INTERLEUKINS-21 AND 22

[0001] This application claims benefit under 35 U.S.C. 19(e) of the filing dates of copending U.S. Provisional Application Serial No. 60/087,340, filed on May 29, 1998, copending U.S. Provisional Application Serial No. 60/099,805, filed on Sep. 10, 1998, and copending U.S. Provisional Application Serial No. 60/131,965, filed on Apr. 30, 1999, each of which is hereby incorporated by reference in its entirety.

[0002] The present invention relates to two novel human genes, each of which encodes a polypeptide which is a member of the Interleukin family. More specifically, the present invention relates to a polynucleotide encoding a novel human polypeptide named Interleukin-21, or IL-21. The present invention also relates to a polynucleotide encoding a novel human polypeptide named Interleukin-22, or IL-22. This invention also relates to IL-21 and IL-22 polypeptides, as well as vectors, host cells, antibodies directed to IL-21 and IL-22 polypeptides, and recombinant methods for producing the same. Also provided are diagnostic methods for detecting disorders related to the immune system, and therapeutic methods for treating such disorders. The invention further relates to screening methods for identifying agonists and antagonists of IL-21 and IL-22 activity.

[0003] Cytokines typically exert their respective biochemical and physiological effects by binding to specific receptor molecules. Receptor binding then stimulates specific signal transduction pathways (Kishimoto, T., et al., Cell 76:253-262 (1994)). The specific interactions of cytokines with their receptors are often the primary regulators of a wide variety of cellular processes including activation, proliferation, and differentiation (Arai, K. -I, et al., Ann. Rev. Biochem. 59:783-836 (1990); Paul, W. E. and Seder, R. A., Cell 76:241-251 (1994)).

[0004] Human interleukin (IL)-17, a closely related homolog of the molecules of the present invention, was only recently identified. IL-17 is a 155 amino acid polypeptide which was molecularly cloned from a CD4 T-cell cDNA library (Yao, Z., et al., J. Immunol. 155:5483-5486 (1995)). The IL-17 polypeptide contains an N-terminal signal peptide and contains approximately 72% identity at the amino acid level with a T-cell trophic herpesvirus saimiri (HVS) gene designated HVS 13. High levels of IL-17 are secreted from CD4-positive primary peripheral blood leukocytes (PBL) upon stimulation (Yao, Z., et al., Immunity 3:811-821 (1995)). Treatment of fibroblasts with IL-17, HVS13, or another murine homologue, designated CTLA8, activate signal transduction pathways and result in the stimulation of the NF-kappaB transcription factor family, the secretion of IL-6, and the costimulation of T-cell proliferation (Yao, Z., et al., Immunity 3:811-821 (1995)).

[0005] An HVS 13-Fc fusion protein was used to isolate a murine IL-17 receptor molecule which does not appear to belong to any of the previously described cytokine receptor families (Yao, Z., et al., Immunity 3:811-821 (1995)). The murine IL-17 receptor (mIL-17R) is predicted to encode a type I transmembrane protein of 864 amino acids with an apparent molecular mass of 97.8 kDa. mIL-17R is predicted to possess an N-terminal signal peptide with a cleavage site between alanine-31 and serine-32. The molecule also contains a 291 amino acid extracellular domain, a 21 amino acid transmembrane domain, and a 521 amino acid cytoplasmic tail. A soluble recombinant WL-17R molecule consisting of 323 amino acids of the extracellular domain of IL-17R fused to the Fc portion of human immunoglobulin IgG1 was able to significantly inhibit IL-17-induced IL-6 production by murine NIH-3T3 cells (supra).

[0006] Interestingly, the expression of the IL-17 gene is highly restricted. It is typically observed primarily in activated T-lymphocyte memory cells (Broxmeyer, H. J. Exp. Med. 183:2411-2415 (1996); Fossiez, F., et al., J. Exp. Med. 183:2593-2603 (1996)). Conversely, the IL-17 receptor appears to be expressed in a large number of cells and tissues (Rouvier, E., et al., J. Immunol. 150:5445-5456 (1993); Yao, Z., et al., J. Immunol. 155:5483-5486 (1995)). It remains to be seen, however, if IL-17 itself can play an autocrine role in the expression of IL-17. IL-17 has been implicated as a causative agent in the expression of IL-6, IL-8, G-CSF, Prostaglandin E (PGE₂), and intracellular adhesion molecule (ICAM)-1 (Fossiez, F., supra; Yao, Z., et al., Immunity 3:811-821 (1995)). Each of these molecules possesses highly relevant and potentially therapeutically valuable properties. For instance, IL-6 is involved in the regulation of hematopoietic stem and progenitor cell growth and expansion (Ikebuchi, K., et al., Proc. Natl. Acad. Sci. USA 84:9035-9039 (1987); Gentile, P. and Broxmeyer, H. E. Ann. N.Y. Acad. Sci. USA 628:74-83 (1991)). IL-8 exhibits a myelosuppressive activity for stem cells and immature subsets of myeloid progenitors (Broxmeyer, H. E., et al., Ann. Hematol. 71:235-246 (1995); Daly, T. J., et al., J. Biol. Chem. 270:23282-23292 (1995)). G-CSF acts both early and late to activate and stimulate hematopoiesis in general, and more specifically on neutrophil hematopoiesis, while PGE₂ enhances erythropoiesis, suppresses lymphopoiesis and myelopoiesis in general, and strongly suppresses monocytopoiesis (Broxmeyer, H. E. Amer. J. Ped. Hematol./Oncol. 14:22-30 (1992); Broxmeyer, H. E. and Williams, D. E. CRC Crit. Rev. Oncol./Hematol. 8:173-226 (1988)).

[0007] Thus, there is a need for polypeptides that function as immunoregulatory molecules and, thereby, modulate the transfer of an extracellular signal ultimately to the nucleus of the cell, since disturbances of such regulation may be involved in disorders relating to cellular activation, hemostasis, angiogenesis, tumor metastasis, cellular migration and ovulation, as well as neurogenesis. Therefore, there is a need for identification and characterization of such human polypeptides which can play a role in detecting, preventing, ameliorating or correcting such disorders.

[0008] The present invention relates to novel polynucleotides and the encoded polypeptides of IL-21 and IL-22. Moreover, the present invention relates to vectors, host cells, antibodies, and recombinant methods for producing the polypeptides and polynucleotides. Also provided are diagnostic methods for detecting disorders related to the polypeptides, and therapeutic methods for treating such disorders. The invention further relates to screening methods for identifying binding partners of IL-21 and IL-22.

[0009] FIG. 1 shows the partial nucleotide sequence (SEQ ID NO:1) and the deduced amino acid sequence (SEQ ID NO:2) of IL-21. The locations of conserved Domains I-IV (see below) are underlined and labeled as such.

[0010] FIGS. 2A and 2B show the nucleotide sequence (SEQ ID NO:3) and the deduced amino acid sequence (SEQ ID NO:4) of IL-22. The locations of conserved Domains I-IV (see below) are underlined and labeled as such. The locations of two potential N-linked glycosylation sites are identified by a bolded asparagine symbol (N) accompanied by a bolded pound sign () located above the initial nucleotide of the codon encoding the corresponding asparagine.

[0011] FIGS. 3A, 3B, and 3C show the regions of identity between the amino acid sequences of: (1) human Interleukin-17 (designated WL-17.aa in the figure; GenBank Accession No. U32659; SEQ ID NO:5); (2) mouse Interleukin-17 (designated mIL-17.aa in the figure; GenBank Accession No. U43088; SEQ ID NO:6); (3) viral Interleukin-17 (designated vIL-17.aa in the figure; GenBank Accession No. X64346; SEQ ID NO:7); (4) IL-20 (designated IL20.aa in the figure and disclosed in copending U.S. Provisional Application Serial No. 60/060,140; filed Sep. 26, 1997; SEQ ID NO:8); (5) a partial-length IL-21 protein (SEQ ID NO:2); (6) the full-length IL-21 protein (designated IL-21FL.aa in the figure); (7) a partial-length IL-22 protein (designated IL-22.aa in the figure), and (8) an IL-22 protein (designated I22ext.aa in the figure), as determined by aligning the sequences using the MegAlign component of the computer program DNA*Star (DNASTAR, Inc., 1228 S. Park St., Madison, Wis. 53715 USA) using the default parameters.

[0012] FIG. 4 shows an analysis of the partial IL-21 amino acid sequence (SEQ ID NO:2). Alpha, beta, turn and coil regions; hydrophilicity and hydrophobicity; amphipathic regions; flexible regions; antigenic index and surface probability are shown. In the Antigenic Index or Jameson-Wolf graph, the positive peaks indicate locations of the highly antigenic regions of the IL-21 protein, that is, regions from which epitope-bearing peptides of the invention can be determined. Polypeptides and polynucleotides encoding polypeptides comprising the domains defined by these graphs are contemplated by the present invention.

[0013] FIG. 5 shows an analysis of the IL-22 amino acid sequence. Alpha, beta, turn and coil regions; hydrophilicity and hydrophobicity; amphipathic regions; flexible regions; antigenic index and surface probability are shown. In the Antigenic Index or Jameson-Wolf graph, the positive peaks indicate locations of the highly antigenic regions of the IL-22 protein, that is, regions from which epitope-bearing peptides of the invention can be determined. Polypeptides and polynucleotides encoding polypeptides comprising the domains defined by these graphs are contemplated by the present invention.

[0014] The data presented in FIG. 5 are also represented in tabular form in Table II. The columns are labeled with the headings Res, Position, and Roman Numerals I-XIII. The column headings refer to the following features of the amino acid sequence presented in FIG. 5 and Table II: Res: amino acid residue of SEQ ID NO:4 or FIGS. 2A and 2B; Position: position of the corresponding residue within SEQ ID NO:4 or FIGS. 2A and 2B; I: Alpha, RegionsGarnier-Robson; II: Alpha, RegionsChou-Fasman; III: Beta, RegionsGarnier-Robson; IV: Beta, RegionsChou-Fasman; V: Turn, RegionsGarnier-Robson; VI: Turn, RegionsChou-Fasman; VII: Coil, RegionsGarnier-Robson; VIII: Hydrophilicity PlotKyte-Doolittle; IX: Alpha, Amphipathic RegionsEisenberg; X: Beta, Amphipathic RegionsEisenberg; XI: Flexible RegionsKarplus-Schulz; XII: Antigenic IndexJameson-Wolf; and XIII: Surface Probability PlotEmini.

[0015] FIGS. 6A and 6B show the nucleotide sequence (SEQ ID NO:28) and the deduced amino acid sequence (SEQ ID NO:29) of the full-length IL-21. The locations of conserved Domains I-IV (identical to those shown in FIG. 1) and of conserved Domains V-VII are underlined and labeled as such. A predicted signal peptide from methionine-1 to alanine-18 is double underlined.

[0016] FIG. 7 shows an analysis of a full-length IL-21 amino acid sequence. Alpha, beta, turn and coil regions; hydrophilicity and hydrophobicity; amphipathic regions; flexible regions; antigenic index and surface probability are shown. In the Antigenic Index or Jameson-Wolf graph, the positive peaks indicate locations of the highly antigenic regions of a full-length IL-21 protein, that is, regions from which epitope-bearing peptides of the invention can be determined. Polypeptides and polynucleotides encoding polypeptides comprising the domains defined by these graphs are contemplated by the present invention.

[0017] The data presented in FIG. 7 are also represented in tabular form in Table I. The columns are labeled with the headings Res, Position, and Roman Numerals I-XIV. The column headings refer to the following features of the amino acid sequence presented in FIG. 7 and Table I: Res: amino acid residue of SEQ ID NO:29 or FIGS. 6A and 6B; Position: position of the corresponding residue within SEQ ID NO:29 or FIGS. 6A and 6B; I: Alpha, RegionsGarnier-Robson; II: Alpha, RegionsChou-Fasman; III: Beta, RegionsGarnier-Robson; IV: Beta, RegionsChou-Fasman; V: Turn, RegionsGarnier-Robson; VI: Turn, RegionsChou-Fasman; VII: Coil, RegionsGarnier-Robson; VIII: Hydrophilicity PlotKyte-Doolittle; IX: Hydrophobicity PlotHopp-Woods; X: Alpha, Amphipathic RegionsEisenberg; XI: Beta, Amphipathic RegionsEisenberg; XII: Flexible RegionsKarplus-Schulz; Xm: Antigenic IndexJameson-Wolf; and XIV: Surface Probability PlotEmini.

[0018] FIG. 8 shows the nucleotide sequence (SEQ ID NO:31) and the deduced amino acid sequence (SEQ ID NO:32) of an IL-22. The locations of conserved Domains I-IV and VI-VII are underlined and labeled as such. The locations of two potential N-linked glycosylation sites are identified by a bolded asparagine symbol (N) accompanied by a bolded pound sign () located above the initial nucleotide of the codon encoding the corresponding asparagine. The two potential N-linked glycosylation sites are located at Asn-39 (N-39, A-40, S-41) and Asn-152 (N-152, S-153, S-154) of SEQ ID NO:32.

[0019] FIG. 9 shows an analysis of the IL-22 amino acid sequence provided in FIG. 8 and SEQ ID NO:32. Alpha, beta, turn and coil regions; hydrophilicity and hydrophobicity; amphipathic regions; flexible regions; antigenic index and surface probability are shown. In the Antigenic Index or Jameson-Wolf graph, the positive peaks indicate locations of the highly antigenic regions of the IL-22 protein, that is, regions from which epitope-bearing peptides of the invention can be determined. Polypeptides and polynucleotides encoding polypeptides comprising the domains defined by these graphs are contemplated by the present invention.

[0020] The data presented in FIG. 9 are also represented in tabular form in Table III. The columns are labeled with the headings Res, Position, and Roman Numerals I-XIV. The column headings refer to the following features of the amino acid sequence presented in FIG. 9 and Table III: Res: amino acid residue of SEQ ID NO:32 or FIG. 8; Position: position of the corresponding residue within SEQ ID NO:32 or FIG. 8; I: Alpha, RegionsGarnier-Robson; II: Alpha, RegionsChou-Fasman; III: Beta, RegionsGarnier-Robson; IV: Beta, RegionsChou-Fasman; V: Turn, RegionsGamier-Robson; VI: Turn, RegionsChou-Fasman; VII: Coil, RegionsGarnier-Robson; VIII: Hydrophilicity PlotKyte-Doolittle; IX: Hydrophobicity PlotHopp-Woods; X: Alpha, Amphipathic RegionsEisenberg; XI: Beta, Amphipathic RegionsEisenberg; XII: Flexible RegionsKarplus-Schulz; XIII: Antigenic IndexJameson-Wolf; and XIV: Surface Probability PlotEmini.

[0021] Definitions

[0022] The following definitions are provided to facilitate understanding of certain terms used throughout this specification.

[0023] In the present invention, isolated refers to material removed from its original environment (e.g., the natural environment if it is naturally occurring), and thus is altered by the hand of man from its natural state. For example, an isolated polynucleotide could be part of a vector or a composition of matter, or could be contained within a cell, and still be isolated because that vector, composition of matter, or particular cell is not the original environment of the polynucleotide. However, a nucleic acid contained in a clone that is a member of a library (e.g., a genomic or cDNA library) that has not been isolated from other members of the library (e.g., in the form of a homogeneous solution containing the clone and other members of the library) or which is contained on a chromosome preparation (e.g., a chromosome spread), is not isolated for the purposes of this invention.

[0024] In the present invention, a secreted IL-21 or IL-22 protein refers to a protein capable of being directed to the ER, secretory vesicles, or the extracellular space as a result of a signal sequence, as well as an IL-21 or IL-22 protein released into the extracellular space without necessarily containing a signal sequence. If the IL-21 or IL-22 secreted protein is released into the extracellular space, the IL-21 or IL-22 secreted protein can undergo extracellular processing to produce a mature IL-21 or IL-22 protein. Release into the extracellular space can occur by many mechanisms, including exocytosis and proteolytic cleavage.

[0025] As used herein, an IL-21 or IL-22 polynucleotide refers to a molecule having a nucleic acid sequence contained in SEQ ID NO:1 or in SEQ ID NO:3, respectively, or the cDNA contained within the respective clones deposited with the ATCC. For example, the IL-21 or IL-22 polynucleotide can contain the nucleotide sequence of the full-length cDNA sequence, including the 5 and 3 untranslated sequences, the coding region, with or without the signal sequence, the secreted protein coding region, as well as fragments, epitopes, domains, and variants of the nucleic acid sequence. Moreover, as used herein, an IL-21 or IL-22 polypeptide refers to a molecule having the translated amino acid sequence generated from the polynucleotide as broadly defined.

[0026] As used herein , an IL-21 polynucleotide refers to a molecule having a nucleic acid sequence contained in SEQ ID NO:1 or in SEQ ID NO:28, or the cDNA contained within the respective clones deposited with the ATCC. For example, the IL-21 polynucleotide can contain the nucleotide sequence of the full-length cDNA sequence, including the 5 and 3 untranslated sequences, the coding region, with or without the signal sequence, the secreted protein coding region, as well as fragments, epitopes, domains, and variants of the nucleic acid sequence. Moreover, as used herein, an IL-21 polypeptide refers to a molecule having the translated amino acid sequence generated from the polynucleotide as broadly defined.

[0027] As used herein , an IL-22 polynucleotide refers to a molecule having a nucleic acid sequence contained in SEQ ID NO:3 or in SEQ ID NO:31, or the cDNA contained within the respective clones deposited with the ATCC. For example, the IL-22 polynucleotide can contain the nucleotide sequence of the full-length cDNA sequence, including the 5 and 3 untranslated sequences, the coding region, with or without the signal sequence, the secreted protein coding region, as well as fragments, epitopes, domains, and variants of the nucleic acid sequence. Moreover, as used herein, an IL-22 polypeptide refers to a molecule having the translated amino acid sequence generated from the polynucleotide as broadly defined.

[0028] A representative clone containing all or most of the sequence for SEQ ID NO:1 (designated HTGED19) was deposited with the American Type Culture Collection (ATCC) on Mar. 5, 1998, and was given the ATCC Deposit Number 209666. In addition, a representative clone containing all or most of the sequence for SEQ ID NO:3 (designated HFPBX96) was also deposited with the ATCC on Mar. 5, 1998, and was given the ATCC Deposit Number 209665. The ATCC is located at 10801 University Blvd., Manassas, Va. 20110-2209, USA. The ATCC deposit was made pursuant to the terms of the Budapest Treaty on the international recognition of the deposit of microorganisms for purposes of patent procedure.

[0029] An IL-21 polynucleotide also includes those polynucleotides capable of hybridizing, under stringent hybridization conditions, to sequences contained in SEQ ID NO:1 or SEQ ID NO:28, the complements thereof, or the cDNA within the deposited clone. Further, an TL-22 polynucleotide also includes those polynucleotides capable of hybridizing, under stringent hybridization conditions, to sequences contained in SEQ ID NO:3 or SEQ ID NO:31, the complements thereof, or the cDNA within the deposited clone. Stringent hybridization conditions refers to an overnight incubation at 42 C. in a solution comprising 50% formamide, 5 SSC (750 mM NaCl, 75 mM sodium citrate), 50 mM sodium phosphate (pH 7.6), 5 Denhardt's solution, 10% dextran sulfate, and 20 g/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1 SSC at about 65 C.

[0030] Also contemplated are nucleic acid molecules that hybridize to the IL-21 and the IL-22 polynucleotides at lower stringency hybridization conditions. Changes in the stringency of hybridization and signal detection are primarily accomplished through the manipulation of formamide concentration (lower percentages of formamide result in lowered stringency); salt conditions, or temperature. For example, lower stringency conditions include an overnight incubation at 37 C. in a solution comprising 6 SSPE (20 SSPE3M NaCl; 0.2M NaH₂PO₄; 0.02M EDTA, pH 7.4), 0.5% SDS, 30% formamide, 100 g/ml salmon sperm blocking DNA; followed by washes at 50 C. with 1 SSPE, 0.1% SDS. In addition, to achieve even lower stringency, washes performed following stringent hybridization can be done at higher salt concentrations (e.g. 5 SSC).

[0031] Note that variations in the above conditions may be accomplished through the inclusion and/or substitution of alternate blocking reagents used to suppress background in hybridization experiments. Typical blocking reagents include Denhardt's reagent, BLOTTO, heparin, denatured salmon sperm DNA, and commercially available proprietary formulations. The inclusion of specific blocking reagents may require modification of the hybridization conditions described above, due to problems with compatibility.

[0032] Of course, a polynucleotide which hybridizes only to polyA sequences (such as any 3 terminal polyA tract of a cDNA shown in the sequence listing), or to a complementary stretch of T (or U) residues, would not be included in the definition of polynucleotide, since such a polynucleotide would hybridize to any nucleic acid molecule containing a polyA stretch or the complement thereof (e.g., practically any double-stranded cDNA clone).

[0033] The EL-21 and IL-22 polynucleotides can be composed of any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. For example, the IL-21 and IL-22 polynucleotides can be composed of single- and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions. In addition, the IL-21 polynucleotides can be composed of triple-stranded regions comprising RNA or DNA or both RNA and DNA. IL-21 polynucleotides may also contain one or more modified bases or DNA or RNA backbones modified for stability or for other reasons. Modified bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications can be made to DNA and RNA; thus, polynucleotide embraces chemically, enzymatically, or metabolically modified forms.

[0034] IL-21 and IL-22 polypeptides can be composed of amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres, and may contain amino acids other than the 20 gene-encoded amino acids. The IL-21 and IL-22 polypeptides may be modified by either natural processes, such as posttranslational processing, or by chemical modification techniques which are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature. Modifications can occur anywhere in the IL-21 and IL-22 polypeptides, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given IL-21 or IL-22 polypeptide. Also, a given IL-21 or IL-22 polypeptide may contain many types of modifications. IL-21 or IL-22 polypeptides may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched, and branched cyclic IL-21 and IL-22 polypeptides may result from posttranslation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination. (See, for instance, PROTEINSSTRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993); POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C. Johnson, Ed., Academic Press, New York, pp. 1-12 (1983); Seifter, et al., Meth. Enzymol. 182:626-646 (1990); Rattan, et al., Ann. NY Acad. Sci. 663:48-62 (1992)).

[0035] SEQ ID NO:1 and SEQ ID NO:28 refer to an IL-21 polynucleotide sequence while SEQ ID NO:2 and SEQ ID NO:29 refer to an IL-21 polypeptide sequence. Likewise, SEQ ID NO:3 and SEQ ID NO:31 refer to an IL-22 polynucleotide sequence while SEQ ID NO:4 and SEQ ID NO:32 refer to an IL-22 polypeptide sequence.

[0036] An IL-21 polypeptide having biological activity refers to polypeptides exhibiting activity similar, but not necessarily identical to, an activity of an IL-21 polypeptide, including mature forms, as measured in a particular biological assay, with or without dose-dependency. In addition, an IL-22 polypeptide having biological activity refers to polypeptides exhibiting activity similar, but not necessarily identical to, an activity of an IL-22 polypeptide, including mature forms, as measured in a particular biological assay, with or without dose-dependency. In the case where dose-dependency does exist, it need not be identical to that of the IL-21 or IL-22 polypeptide, but rather substantially similar to the dose-dependence in a given activity as compared to the IL-21 or IL-22 polypeptides (i.e., the candidate polypeptide will exhibit greater activity or not more than about 25-fold less and, preferably, not more than about tenfold less activity, and most preferably, not more than about three-fold less activity relative to the IL-21 polypeptide).

[0037] IL-21 and IL-22 Polynucleotides and Polypeptides

[0038] Clone HTGED19, encoding IL-21, was isolated from a cDNA library derived from apoptotic T-cells. This clone contains the entire coding region identified as SEQ ID NO:2. The deposited clone contains a cDNA having a total of 705 nucleotides, which encodes a partial predicted open reading frame of 87 amino acid residues (see FIG. 1). The partial open reading frame begins at a point in the complete IL-21 ORF such that the G in position 1 of SEQ ID NO:1 is actually in position 3 of a coding triplet. As such, the partial predicted IL-21 polypeptide sequence is shown beginning in-frame with an alanine residue at position 1 of SEQ ID NO:2. The alanine residue at position 1 of SEQ ID NO:2 is encoded by nucleotides 2-4 of the nucleotide sequence shown as SEQ ID NO:1. The ORF shown as SEQ ID NO:2 ends at a stop codon at nucleotide position 263-265 of the nucleotide sequence shown as SEQ ID NO:1. The predicted molecular weight of the partial IL-21 protein should be about 9,558 Daltons.

[0039] An initial BLAST analysis of the expression of the IL-21 cDNA sequence against the HGS EST database has also revealed a highly specific expression of this cDNA clone. In such an analysis, the HTGED19 cDNA sequence appears to be found only in apoptotic T-cells. Thus, IL-21 appears to be expressed in a highly restricted pattern limited to apoptotic T-cells, and, for example, other subpopulatons of lymphocytes or other cells in a state of activation or quiescence.

[0040] Clone HTGED19, encoding IL-21, was used to screen a panel of bacterial artificial chromosomes containing various segments of human genomic DNA (Research Genetics, Inc.). A positive clone was sequenced to identify potential splice donor and acceptor sites. Analysis of several sites revealed an upstream partial ORF that, when placed immediately 5 and in frame with the existing IL-21 DNA sequence, generated a complete ORF which encodes a polypeptide with additional sequence identity to the IL-17 family (See FIGS. 3A, 3B, and 3C). A clone of the full-length IL-21 ORF has been constructed by combination of the IL-21 exons PCR-amplified from the HTGED19 genomic clone. The clone has been deposited with the ATCC as ATCC Deposit No. PTA-69 on May 14, 1999. The nucleotide sequence of the full-length IL-21 clone contains the entire coding region identified as SEQ ID NO:29. The resultant clone contains an insert having a total of 1067 nucleotides, which encodes a predicted open reading frame of 197 amino acid residues (see FIGS. 6A and 6B). The open reading frame begins at nucleotide position 34 in the complete IL-21 polynucleotide shown as SEQ ID NO:28 (FIGS. 6A and 6B). The ORF ends at a stop codon at nucleotide position 625-627 of the nucleotide sequence shown as SEQ ID NO:28 (FIGS. 6A and 6B). The predicted molecular weight of the IL-21 polypeptide shown in FIGS. 6A and 6B and as SEQ ID NO:29 should be about 21,764 Daltons.

[0041] Further BLAST analysis of the expression of the full-length IL-21 cDNA sequence against the HGS EST database has also revealed a highly specific expression of this cDNA clone. In such an analysis, the full-length HTGED19 cDNA sequence appears to be found only in apoptotic T-cells. Thus, IL-21 appears to be expressed in a highly restricted pattern limited to apoptotic T-cells, and, for example, other subpopulatons of lymphocytes or other cells in a state of activation or quiescence.

[0042] A PCR product comprising exons 1 and 2 (based on the genomic organization predicted above) has been amplified using a 12 week old early stage cDNA library as template DNA. This PCR product confirms that at least exons 1 and 2 of the genomic organization predicted above exists as messenger RNA in at least 12 week old early stage human embyo.

[0043] Clone HFPBX96, encoding IL-22, was isolated from a cDNA library derived from epileptic frontal cortex. This clone contains the entire coding region identified as SEQ ID NO:4. The deposited clone contains a cDNA having a total of 1,642 nucleotides, which encodes a partial predicted open reading frame of 160 amino acid residues (see FIGS. 2A and 2B). The partial open reading frame begins at a point in the complete IL-22 ORF such that the G in position 1 of SEQ ID NO:3 is actually in position two of a coding triplet. As such, the partial predicted IL-22 polypeptide sequence is shown beginning in-frame with an asparagine residue at position 1 of SEQ ID NO:4. The asparagine residue at position 1 of SEQ ID NO:4 is encoded by nucleotides 3-5 of the nucleotide sequence shown as SEQ ID NO:3. The ORF shown as SEQ ID NO:4 ends at a stop codon at nucleotide position 483-485 of the nucleotide sequence shown as SEQ ID NO:3. The predicted molecular weight of the partial IL-22 protein should be about 17,436 Daltons.

[0044] Clone HFPBX96, encoding IL-22, was used to screen a human fetal brain cDNA library containing approximately one million cDNA clones (Genome Systems, Inc.). A positive clone was sequenced to identify 59 nucleotides of additional 5 sequence. The cDNA clone has been deposited with the ATCC as ATCC Deposit No. PTA-70 on May 14, 1999. Analysis of the extended IL-22 ORF reveals a polypeptide with additional sequence identity to the IL-17 family (see FIGS. 3A, 3B, and 3C). The nucleotide sequence of the extended, but still apparently partial-length IL-22 clone contains the entire coding region identified as SEQ ID NO:31. The resultant clone contains an insert having a total of 522 nucleotides, which encodes a predicted open reading frame of 174 amino acid residues (see FIG. 8). The open reading frame begins at nucleotide position 1 in the complete IL-22 polynucleotide shown as SEQ ID NO:31 (FIG. 8). The ORF ends at a stop codon at nucleotide position 520-522 of the nucleotide sequence shown as SEQ ID NO:31 (FIG. 8). The predicted molecular weight of the IL-22 polypeptide shown in FIG. 8 and as SEQ ID NO:31 is about 19,636 Daltons.

[0045] Using BLAST and MegAlign analyses, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:29, and SEQ ID NO:32 were each found to be highly homologous to several members of the Interleukin family. Particularly, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:29, and SEQ ID NO:32 contain at least four domains homologous to the translation products of the human mRNA for Interleukin (IL)-20 (copending U.S. Provisional Application Serial No. 60/060,140; filed Sep. 26, 1997; SEQ ID NO:8), IL-17 (GenBank Accession No. U32659; SEQ ID NO:5; see also FIGS. 3A, 3B, and 3C), the murine mRNA for Interleukin (IL)-17 (GenBank Accession No. U43088; SEQ ID NO:6; see also FIGS. 3A, 3B, and 3C), and the human viral mRNA for Interleukin (IL)-17 (GenBank Accession No. X64346; SEQ ID NO:7; see also FIGS. 3A, 3B, and 3C).

[0046] Specifically, the molecules of the present invention, in particular, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:29, and SEQ ID NO:32 share a high degree of sequence identity with IL-20, IL-17, mIL-17, and vIL-17 in the following conserved domains: (a) a predicted NXDPXRYP domain (where X represents any amino acid) located at about amino acids valine-3 to proline-11 of SEQ ID NO:2, serine-57 to proline-64 of SEQ ID NO:4, valine-113 to proline-121 of SEQ ID NO:29, serine-70 to proline-77 of SEQ ID NO:32, and asparagine-79 to proline-86 of the human IL-17 amino acid sequence (SEQ ID NO:5); (b) a predicted CLCXGC domain (where X represents any amino acid) located at about amino acids cysteine-19 to cysteine-24 of SEQ ID NO:2, cysteine-72 to cysteine-77 of SEQ ID NO:4, cysteine-129 to cysteine-134 of SEQ ID NO:29, cysteine-85 to cysteine-90 of SEQ ID NO:32, and cysteine-94 to cysteine-99 of the human IL-17 amino acid sequence (SEQ ID NO:5); (c) a predicted LVLRRXP domain (where X represents any amino acid) located at about amino acids leucine-46 to proline-52 of SEQ ID NO:2, valine-99 to proline-105 of SEQ ID NO:4, leucine-156 to proline-162 of SEQ ID NO:29, valine-12 to proline-118 of SEQ ID NO:32, and leucine-120 to proline-126 of the human IL-17 amino acid sequence (SEQ ID NO:5); and (d) a predicted VXVGCTCV domain (where X represents any amino acid) located at about amino acids valine-75 to valine-82 of SEQ ID NO:2, isoleucine-121 to valine-128 of SEQ ID NO:4, valine-187 to valine-192 of SEQ ID NO:29, isoleucine-134 to valine-141 of SEQ ID NO:32, and valine-140 to valine-147 of the human IL-17 amino acid sequence (SEQ ID NO:5).

[0047] In addition, the full-length IL-21 molecule shown in FIGS. 6A and 6B (SEQ ID NO:29) and the IL-22 molecule shown in FIG. 8 (SEQ ID NO:32) exhibit several additional conserved domains when compared with IL-20 and the other members of the IL-17 family as shown in FIGS. 3A, 3B, and 3C). These conserved Domains are underlined in FIGS. 6A and 6B and in FIG. 8 and are labeled as conserved Domains V, VI, and VII. Specifically, the molecules of the present invention, in particular, SEQ ID NO:29 and SEQ ID NO:32, share a high degree of sequence identity with IL-20, IL-17, mIL-17, and vIL-17 in the following conserved domains: (a) a predicted PXCXSAE domain (where X represents any amino acid) located at about amino acids proline-34 to glutamic acid-40 of SEQ ID NO:29; (b) a predicted PXXLVS domain (where X represents any amino acid) located at about amino acids proline-63 to serine-68 of SEQ ID NO:29 and at about amino acids alanine-18 to serine-23 of SEQ ID NO:32; and (c) a predicted RSXSPW domain (where X represents any amino acid) located at about amino acids arginine-104 to tryptophan-109 of SEQ ID NO:29 and at about amino acids arginine-60 to tryptophan-65 of SEQ ID NO:32. These polypeptide fragments of IL-21 and IL-22 are specifically contemplated in the present invention. Because each of these IL-17 and IL-17-like molecules is thought to be important immunoregulatory molecules, the homology between these IL-17 and IL-17-like molecules and IL-21 and IL-22 suggests that IL-21 and IL-22 may also be important immunoregulatory molecules.

[0048] Moreover, based on their apparent sequence identities with IL-17 and IL-20 (see FIGS. 3A, 3B, and 3C), the full-length IL-21 and IL-22 polypeptides are each likely to have an amino terminal secretory signal peptide leader sequence. Since the present invention appears to be partial cDNA clones of the IL-21 (SEQ ID NOs:1 and 2) and IL-22 (SEQ ID NOs:3 and 4) molecules (in addition to the full-length IL-21 molecule shown as SEQ ID NOs:28 and 29 and the IL-22 molecule shown as SEQ ID NOs:31 and 32), it is also contemplated that the translation products of SEQ ID NOs:2, 4, and 32 of the present invention will be caused to enter the cellular secretory pathway by virtue of being expressed as a fusion proteins comprising several different portions of the N-terminus of the IL-20 molecule of copending U.S. Provisional Application Serial No. 60/060,140 fused to the known coding sequence of the IL-21 or IL-22 molecules of the present invention. Such expression constructs will secrete hybrid IL-20/IL-21 or IL-20/IL-22 molecules from the host cell.

[0049] In one embodiment, the mature IL-21 protein used in these fusion proteins encompasses about amino acids 12-87 of SEQ ID NO:2, while the IL-20/21 fusion protein encompasses about the 104 or 113 N-terminal amino acids of IL-20 encoded in frame with about amino acids 12-87 of the IL-21 of SEQ ID NO:2. In other embodiments, an IL-20/21 fusion protein encompasses about the 104 or 113 N-terminal amino acids of IL-20 encoded in frame with about amino acids 3-87 of the IL-21 protein of SEQ ID NO:2. These polypeptide fragments of IL-21 are specifically contemplated in the present invention.

[0050] In another embodiment, the mature IL-22 protein used to generate these fusion proteins encompasses about amino acids 1-160 of SEQ ID NO:4, while the IL-20/22 fusion protein encompasses about the 95, 104 or 113 N-terminal amino acids of IL-20 encoded in frame with about amino acids 1-160 of the IL-22 of SEQ ID NO:4. In other embodiments, the IL-22 protein used to generate these fusion proteins encompasses about amino acids 47-160 of SEQ ID NO:4, while the IL-20/22 fusion protein encompasses about the 95, 104 or 113 N-terminal amino acids of IL-20 encoded in frame with about amino acids 1-160 of the IL-22 of SEQ ID NO:4. In still other embodiments, the IL-22 protein used to generate these fusion proteins encompasses about amino acids 56-160 of SEQ ID NO:4, while the IL-20/22 fusion protein encompasses about the 95, 104 or 113 N-terminal amino acids of IL-20 encoded in frame with about amino acids 1-160 of the IL-22 of SEQ ID NO:4. In yet other embodiments, the IL-22 protein used to generate these fusion proteins encompasses about amino acids 65-160 of SEQ ID NO:4, while the IL-20/22 fusion protein encompasses about the 95, 104 or 113 N-terminal amino acids of IL-20 encoded in frame with about amino acids 1-160 of the IL-22 of SEQ ID NO:4. These polypeptide fragments of IL-22 are specifically contemplated in the present invention.

[0051] In yet another embodiment, the mature IL-22 protein used to generate these fusion proteins encompasses about amino acids 1-173 of SEQ ID NO:32, while the IL-20/22 fusion protein encompasses about the 95, 104 or 113 N-terminal amino acids of IL-20 encoded in frame with about amino acids 1-173 of the IL-22 of SEQ ID NO:32. These polypeptide fragments of IL-22 are specifically contemplated in the present invention.

[0052] The IL-21 and IL-22 nucleotide sequences identified as SEQ ID NO:1 and SEQ ID NO:3, respectively, were assembled from partially homologous (overlapping) sequences obtained from the deposited clones. The IL-21 nucleotide sequence identified as SEQ ID NO:28 was assembled from partially homologous (overlapping) sequences obtained from the deposited clone and a genomic DNA clone. The IL-22 nucleotide sequence identified as SEQ ID NO:32 was assembled from partially homologous (overlapping) sequences obtained from the deposited clones (ATCC Deposit No. 209665 and ATCC Deposit No. PTA-70). The overlapping sequences specific to the partial IL-21 and IL-22 molecules of the invention and the full-length IL-21 molecule of the invention were each assembled into single contiguous sequences of high redundancy (usually three to five overlapping sequences at each nucleotide position), resulting in four final sequences identified as SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:28, and SEQ ID NO:31.

[0053] Therefore, SEQ ID NO:1 and the translated SEQ ID NO:2; SEQ ID NO:3 and the translated SEQ ID NO:4; SEQ ID NO:31 and the translated SEQ ID NO:32; and SEQ ID NO:28 and the translated SEQ ID NO:29, are sufficiently accurate and otherwise suitable for a variety of uses well known in the art and described further below. For instance, SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:28, and SEQ ID NO:31 are useful for designing nucleic acid hybridization probes that will detect nucleic acid sequences contained in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:28, and SEQ ID NO:31, or the cDNA contained in the respective deposited cDNA clones. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling a variety of forensic and diagnostic methods of the invention. Similarly, polypeptides identified from SEQ ID NO:2 and SEQ ID NO:29 may be used to generate antibodies which bind specifically to IL-21 and polypeptides identified from SEQ ID NO:4 and SEQ ID NO:32 may be used to generate antibodies which bind specifically to IL-22.

[0054] Nevertheless, DNA sequences generated by sequencing reactions can contain sequencing errors. The errors exist as misidentified nucleotides, or as insertions or deletions of nucleotides in the generated DNA sequence. The erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence. In these cases, the predicted amino acid sequence diverges from the actual amino acid sequence, even though the generated DNA sequence may be greater than 99.9% identical to the actual DNA sequence (for example, one base insertion or deletion in an open reading frame of over 1000 bases).

[0055] Accordingly, for those applications requiring precision in the nucleotide sequence or the amino acid sequence, the present invention provides not only the generated nucleotide sequence identified as SEQ ID NO:1 and the predicted translated amino acid sequence identified as SEQ ID NO:2, and the generated nucleotide sequence identified as SEQ ID NO:28 and the predicted translated amino acid sequence identified as SEQ ID NO:29, but also a sample of plasmid DNA containing a human cDNA of IL-21 deposited with the ATCC. In addition, the present invention also provides not only the generated nucleotide sequence identified as SEQ ID NO:3 and the predicted translated amino acid sequence identified as SEQ ID NO:4, and the generated nucleotide sequence identified as SEQ ID NO:3 and the predicted translated amino acid sequence identified as SEQ ID NO:4, but also a sample of plasmid DNA containing a human cDNA of IL-22 deposited with the ATCC. Accordingly, the nucleotide sequence of the deposited EL-21 and IL-22 clones can be readily determrined by sequencing the deposited clone in accordance with known methods. The predicted IL-21 and IL-22 amino acid sequences can then be verified from such deposits. Moreover, the amino acid sequence of the protein encoded by the deposited clone can also be directly determined by peptide sequencing or by expressing the protein in a suitable host cell containing the deposited human IL-21 or IL-22 cDNAs, collecting the protein, and determining its sequence.

[0056] The present invention also relates to the IL-21 gene corresponding to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:28, SEQ ID NO:29 or the deposited clone which encodes a partial IL-21. The present invention further relates to the IL-22 gene corresponding to SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:31, SEQ ID NO:32 or the deposited clone which encodes IL-22. The IL-21 and IL-22 genes can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include preparing probes or primers from the disclosed sequences and identifying or amplifying the IL-21 and IL-22 genes from appropriate sources of genomic material.

[0057] Also provided in the present invention are species homologs of IL-21 and IL-22. Species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for the desired homolog.

[0058] The IL-21 and IL-22 polypeptides can be prepared in any suitable manner. Such polypeptides include isolated naturally occurring polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods. Means for preparing such polypeptides are well understood in the art.

[0059] The IL-21 and IL-22 polypeptides may be in the form of the secreted protein, including the mature form, or may be a part of a larger protein, such as a fusion protein. It is often advantageous to include an additional amino acids which comprise secretory or leader sequences, pro-sequences, sequences which aid in purification, such as multiple histidine residues, or an additional sequence for stability during recombinant production.

[0060] IL-21 and IL-22 polypeptides are preferably provided in an isolated form, and preferably are substantially purified. A recombinantly produced version of an IL-21 or IL-22 polypeptide, including the secreted polypeptide, can be substantially purified by the one-step method described in the publication by Smith and Johnson (Gene 67:31-40 (1988)). IL-21 and EL-22 polypeptides also can be purified from natural or recombinant sources using antibodies of the invention raised against the IL-21 and IL-22 proteins, respectively, in methods which are well known in the art.

[0061] Polynucleotide and Polypeptide Variants

[0062] Variant refers to a polynucleotide or polypeptide differing from the IL-21 and IL-22 polynucleotides or polypeptides, but retaining essential properties thereof. Generally, variants are overall closely similar, and, in many regions, identical to the IL-21 and IL-22 polynucleotide or polypeptide.

[0063] By a polynucleotide having a nucleotide sequence at least, for example, 95% identical to a reference nucleotide sequence of the present invention, it is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence encoding the IL-21 or IL-22 polypeptides. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be inserted, deleted or substituted with another nucleotide. The query sequence may be an entire sequence shown of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:28, SEQ ID NO:31, the ORF (open reading frame) of either IL-21 or IL-22, or any fragment specified as described herein.

[0064] As a practical matter, whether any particular nucleic acid molecule or polypeptide is at least 90%, 95%, 96%, 97%, 98% or 99% identical to (or 10%, 5%, 4%, 3%, 2% or 1% different from) a nucleotide sequence of the presence invention can be determined conventionally using known computer programs. A preferred method for determining the best overall match between a query sequence (a sequence of the present invention) and a subject sequence, also referred to as a global sequence alignment, can be determined using the FASTDB computer program based on the algorithm of Brutlag and colleagues (Comp. App. Biosci. 6:237-245 (1990)). In a sequence alignment the query and subject sequences are both DNA sequences. An RNA sequence can be compared by converting (uridine residues (U) to thymidine residues (T). The result of said global sequence alignment is in percent identity. Preferred parameters used in a FASTDB alignment of DNA sequences to calculate percent identiy are: MatrixUnitary, k-tuple4, Mismatch Penalty1, Joining Penalty30, Randomization Group Length0, Cutoff Scorel, Gap Penalty5, Gap Size Penalty 0.05, Window Size500 or the length of the subject nucleotide sequence, whichever is shorter.

[0065] If the subject sequence is shorter than the query sequence because of 5 or 3 deletions, but not because of internal deletions, a manual correction must be made to the results. This is because the FASTDB algorithm does not account for 5 and 3 truncations of the subject sequence when calculating percent identity. For subject sequences truncated at the 5 or 3 ends, relative to the the query sequence, the percent identity is corrected by calculating the number of bases of the query sequence that are 5 and 3 of the subject sequence, which are not matched/aligned, as a percent of the total bases of the query sequence. Whether a nucleotide is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This corrected score is what is used for the purposes of the present invention. Only bases outside the 5 and 3 bases of the subject sequence, as displayed by the FASTDB alignment, which are not matched/aligned with the query sequence, are calculated for the purposes of manually adjusting the percent identity score.

[0066] For example, a 90 base subject sequence is aligned to a 100 base query sequence to determine percent identity. The deletions occur at the 5 end of the subject sequence and therefore, the FASTDB alignment does not show a matched/alignment of the first 10 bases at 5 end. The 10 unpaired bases represent 10% of the sequence ((number of bases at the 5 and 3 ends not matched)/(total number of bases in the query sequence)), so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 bases were perfectly matched the final percent identity would be 90%. In another example, a 90 base subject sequence is compared with a 100 base query sequence. This time the deletions are internal deletions so that there are no bases on the 5 or 3 of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only bases 5 and 3 of the subject sequence which are not matched/aligned with the query sequnce are manually corrected for. No other manual corrections are to made for the purposes of the present invention.

[0067] By a polypeptide having an amino acid sequence which is, at least, for example, 95% identical to (or 5% different from) a query amino acid sequence of the present invention, it is intended that the amino acid sequence of the subject polypeptide is identical to the query sequence except that the subject polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the query amino acid sequence. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a query amino acid sequence, up to 5% of the amino acid residues in the subject sequence may be inserted, deleted, (insertions and deletions are collectively referred to as indels in the art) or substituted with another amino acid. These alterations of the reference sequence may occur at the amino- or carboxy-terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence.

[0068] As a practical matter, whether any particular polypeptide is at least 90%, 95%, 96%, 97%, 98% or 99% identical to (or 10%, 5%, 4%, 3%, 2% or 1% different from), for instance, the amino acid sequences shown in SEQ ID NO:2 or SEQ ID NO:29, or that shown in SEQ ID NO:4 or SEQ ID NO:32, or to the amino acid sequence encoded by deposited cDNA clones, can be determined conventionally using known computer programs. A preferred method for determing the best overall match between a query sequence (a sequence of the present invention) and a subject sequence, also referred to as a global sequence alignment, can be determined using the FASTDB computer program based on the algorithm of Brutlag and colleagues (Comp. App. Biosci. 6:237-245 (1990)). In a sequence alignment the query and subject sequences are either both nucleotide sequences or both amino acid sequences. The result of said global sequence alignment is in percent identity. Preferred parameters used in a FASTDB amino acid alignment are: MatrixPAM 0, k-tuple2, Mismatch Penalty1, Joining Penalty20, Randomization Group Length0, Cutoff Score1, Window Sizesequence length, Gap Penalty5, Gap Size Penalty0.05, Window Size500 or the length of the subject amino acid sequence, whichever is shorter.

[0069] If the subject sequence is shorter than the query sequence due to N- or C-terminal deletions, not because of internal deletions, a manual correction must be made to the a results. This is becuase the FASTDB program does not account for N- and C-terminal truncations of the subject sequence when calculating global percent identity. For subject sequences truncated at the N- and C-terrmini, relative to the the query sequence, the percent identity is corrected by calculating the number of residues of the query sequence that are N- and C-terminal of the subject sequence, which are not matched/aligned with a corresponding subject residue, as a percent of the total bases of the query sequence. Whether a residue is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This final percent identity score is what is used for the purposes of the present invention. Only residues to the N- and C-termini of the subject sequence, which are not matched/aligned with the query sequence, are considered for the purposes of manually adjusting the percent identity score. That is, only query residue positions outside the farthest N- and C-terminal residues of the subject sequence.

[0070] For example, a 90 amino acid residue subject sequence is aligned with a 100 residue query sequence to determine percent identity. The deletion occurs at the N-terminus of the subject sequence and therefore, the FASTDB alignment does not show a matching/alignment of the first 10 residues at the N-terminus. The 10 unpaired residues represent 10% of the sequence (number of residues at the N- and C-termini not matched/total number of residues in the query sequence), so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 residues were perfectly matched the final percent identity would be 90%. In another example, a 90 residue subject sequence is compared with a 100 residue query sequence. This time the deletions are internal deletions so there are no residues at the N- or C-termini of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only residue positions outside the N- and C-terminal ends of the subject sequence, as displayed in the FASTDB alignment, which are not matched/aligned with the query sequnce are manually corrected for. No other manual corrections are to made for the purposes of the present invention.

[0071] The IL-21 and IL-22 variants may contain alterations in the coding regions, non-coding regions, or both. Especially preferred are polynucleotide variants containing alterations which produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded polypeptide. Nucleotide variants produced by silent substitutions due to the degeneracy of the genetic code are preferred. Moreover, variants in which 5-10, 1-5, or 1-2 amino acids are substituted, deleted, or added in any combination are also preferred. IL-21 and IL-22 polynucleotide variants can be produced for a variety of reasons, e.g., to optimize codon expression for a particular host (change codons in the human mRNA to those preferred by a bacterial host such as E. coli).

[0072] Naturally occurring IL-21 and IL-22 variants are called allelic variants, and refer to one of several alternate forms of a gene occupying a given locus on a chromosome of an organism (Genes II, Lewin, B., ed., John Wiley & Sons, New York (1985)). These allelic variants can vary at either the polynucleotide and/or polypeptide level. Alternatively, non-naturally occurring variants may be produced by mutagenesis techniques or by direct synthesis.

[0073] Using known methods of protein engineering and recombinant DNA technology, variants may be generated to improve or alter the characteristics of the IL-21 and IL-22 polypeptides. For instance, one or more amino acids can be deleted from the N-terminus or C-terminus of the secreted protein without substantial loss of biological function. Ron and coworkers reported variant KGF proteins having heparin binding activity even after deleting 3, 8, or 27 amino-terminal amino acid residues (J. Biol. Chem. 268:2984-2988 (1993)). Similarly, Interferon gamama exhibited up to ten times higher activity after deleting 8-10 amino acid residues from the carboxy terminus of this protein (Dobeli, et al., J. Biotechnol. 7:199-216 (1988)).

[0074] In the present case, since the IL-21 and IL-22 proteins of the invention are highly related to the Interleukin-17-like polypeptide family, deletions of N-terminal amino acids up to the cysteine at position 19 of SEQ ID NO:2 and up to the cysteine at position 29 of SEQ ID NO:4 may retain some biological activity. Polypeptides having further N-terminal deletions including the cysteine-19 residue in SEQ ID NO:2 and the cysteine-29 residue in SEQ ID NO:4 would not be expected to retain such biological activities because it is likely that these residues are required for forming a disulfide bridge to provide structural stability which is needed for receptor binding and signal transduction.

[0075] However, even if deletion of one or more amino acids from the N-terminus of a protein results in modification or loss of one or more biological functions of the protein, other biological activities may still be retained. Thus, the ability of the shortened protein to induce and/or bind to antibodies which recognize the complete or mature IL-21 or IL-22 proteins generally will be retained when less than the majority of the residues of the complete or mature IL-21 or IL-22 proteins are removed from the N-termini of the respective proteins. Whether a particular polypeptide lacking N-terminal residues of a complete protein retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art.

[0076] Accordingly, the present invention further provides polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of the IL-21 polypeptide shown in SEQ ID NO:2, up to the cysteine residue at position number 19, and polynucleotides encoding such polypeptides. In addition, the present invention further provides polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of the IL-22 polypeptide shown in SEQ ID NO:4, up to the cysteine residue at position number 29, and polynucleotides encoding such polypeptides. In particular, the present invention provides polypeptides comprising the amino acid sequence of residues n¹-87 of SEQ ID NO:2, where n¹ is an integer in the range of 1 to 18, and 19 is the position of the first residue from the N-terminus of the complete EL-21 polypeptide (shown in SEQ ID NO:2) believed to be required for the receptor binding activity of the IL-21 protein. Likewise, the present invention provides polypeptides comprising the amino acid sequence of residues n²-160 of SEQ ID NO:4, where n² is an integer in the range of 1 to 28, and 29 is the position of the first residue from the N-terminus of the complete IL-22 polypeptide (shown in SEQ ID NO:4) believed to be required for the receptor binding activity of the IL-22 protein.

[0077] More in particular, the invention provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues 1-87, 2-87, 3-87, 4-87, 5-87, 6-87, 7-87, 8-87, 9-87, 10-87, 11-87, 12-87, 13-87, 14-87, 15-87, 16-87, 17-87, 18-87, and 19-87 of SEQ ID NO:2. Polypeptides encoded by these polynucleotides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-21 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0078] The invention also provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues 1-160, 2-160, 3-160, 4-160, 5-160, 6-160, 7-160, 8-160, 9-160, 10-160, 11-160, 12-160, 13-160, 14-160, 15-160, 16-160, 17-160, 18-160, 19-160, 20-160, 21-160, 22-160, 23-160, 24-160, 25-160, 26-160, 27-160, 28-160, and 29-160 of SEQ ID NO:4. Polypeptides encoded by these polynucleotides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-22 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0079] In addition, since the IL-21 and IL-22 proteins of the invention are highly related to the IL-17-like polypeptide family, deletions of C-terminal amino acids up to the leucine at postion 83 of SEQ ID NO:2 and up to the proline at position 129 of SEQ ID NO:4 may retain some biological activity. Polypeptides having further C-terminal deletions including the leucine residue at position 83 of SEQ ID NO:2 and the proline at position 129 of SEQ ID NO:4 would not be expected to retain such biological activities since these residues are in the beginning of the conserved domain required for biological activities.

[0080] However, even if deletion of one or more amino acids from the C-terminus of a protein results in modification of loss of one or more biological functions of the protein, other biological activities may still be retained. Thus, the ability of the shortened protein to induce and/or bind to antibodies which recognize the complete or mature IL-21 and IL-22 proteins generally will be retained when less than the majority of the residues of the complete or mature IL-21 and IL-22 proteins are removed from the C-terminus. Whether a particular polypeptide lacking C-terminal residues of a complete protein retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art.

[0081] Accordingly, the present invention further provides polypeptides having one or more residues removed from the carboxy terminus of the amino acid sequence of the IL-21 polypeptide shown in SEQ ID NO:2, up to the leucine residue at position 83 of SEQ ID NO:2, and polynucleotides encoding such polypeptides. In addition, the present invention further provides polypeptides having one or more residues removed from the carboxy terminus of the amino acid sequence of the IL-22 polypeptide shown in SEQ ID NO:4, up to the proline residues at position 129 of SEQ ID NO:4. In particular, the present invention provides polypeptides having the amino acid sequence of residues 1-m¹ of the amino acid sequence in SEQ ID NO:2, where m¹ is any integer in the range of 83 to 87, and residue 82 is the position of the first residue from the C-terminus of the complete IL-21 polypeptide (shown in SEQ ID NO:2) believed to be required for activity of the IL-21 protein. In addition, the present invention also provides polypeptides having the amino acid sequence of residues 1-m² of the amino acid sequence in SEQ ID NO:4, where m² is any integer in the range of 129 to 160, and residue 128 is the position of the first residue from the C-terminus of the complete IL-22 polypeptide (shown in SEQ ID NO:4) believed to be required for activity of the IL-22 protein.

[0082] More in particular, the invention provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues 1-83, 1-84, 1-85, 1-86, and 1-87 of SEQ ID NO:2. Polypeptides encoded by these polynucleotides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-21 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0083] The present invention also provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues 1-129, 1-130, 1-131, 1-132, 1-133, 1-134, 1-135, 1-136, 1-137, 1-138, 1-139, 1-140, 1-141, 1-142, 1-143, 1-144, 1-145, 1-146, 1-147, 1-148, 1-149, 1-150, 1-151, 1-152, 1-153, 1-154, 1-155, 1-156, 1-157, 1-158, 1-159, and 1-160 of SEQ ID NO:4. Polypeptides encoded by these polynucleotides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-22 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0084] The invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini of IL-21, which may be described generally as having residues n¹-m¹ of SEQ ID NO:2, where n¹ and m¹ are integers as described above. Likewise, the invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini of IL-22, which may be described generally as having residues n²-m² of SEQ ID NO:4, where n² and m² are integers as described above.

[0085] Moreover, ample evidence demonstrates that variants often retain a biological activity similar to that of the naturally occurring protein. For example, Gayle and coworkers conducted extensive mutational analysis of human cytokine IL-1a (J. Biol. Chem. 268:22105-22111 (1993)). They used random mutagenesis to generate over 3,500 individual IL-1a mutants that averaged 2.5 amino acid changes per variant over the entire length of the molecule. Multiple mutations were examined at every possible amino acid position. The investigators found that most of the molecule could be altered with little effect on either binding or biological activity (see, Abstract). In fact, only 23 unique amino acid sequences, out of more than 3,500 nucleotide sequences examined, produced a protein that significantly differed in activity from wild-type.

[0086] Furthermore, even if deleting one or more amino acids from the N-terminus or C-terminus of a polypeptide results in modification or loss of one or more biological functions, other biological activities may still be retained. For example, the ability of a deletion variant to induce and/or to bind antibodies which recognize the secreted form will likely be retained when less than the majority of the residues of the secreted form are removed from the N-terminus or C-terminus. Whether a particular polypeptide lacking N- or C-terminal residues of a protein retains such immunogenic activities can readily be determined by routine methods described herein and otherwise known in the art.

[0087] As mentioned above, even if deletion of one or more amino acids from the N-terminus of a protein results in modification or loss of one or more biological functions of the protein, other biological activities may still be retained. Thus, the ability of the shortened protein to induce and/or bind to antibodies which recognize the complete or mature IL-21 or IL-22 proteins generally will be retained when less than the majority of the residues of the complete or mature IL-21 or IL-22 proteins are removed from the N-termini of the respective proteins. Whether a particular polypeptide lacking N-terminal residues of a complete protein retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art.

[0088] Accordingly, the present invention further provides polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of the IL-21 polypeptide shown in SEQ ID NO:2, up to the valine residue at position number 82, and polynucleotides encoding such polypeptides. In addition, the present invention further provides polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of the IL-22 polypeptide shown in SEQ ID NO:4, up to the aspartic acid residue at position number 155, and polynucleotides encoding such polypeptides. In particular, the present invention provides polypeptides comprising the amino acid sequence of residues n³-87 of SEQ ID NO:2, where n³ is an integer in the range of 1 to 82, and 83 is the position of the first residue from the N-terminus of the complete IL-21 polypeptide (shown in SEQ ID NO:2) believed to be required for immunogenic activity of the IL-21 protein. Likewise, the present invention provides polypeptides comprising the amino acid sequence of residues n⁴-160 of SEQ ID NO:4, where n⁴ is an integer in the range of 1 to 155, and 156 is the position of the first residue from the N-terminus of the complete IL-22 polypeptide (shown in SEQ ID NO:4) believed to be required for immunogenic activity of the IL-22 protein.

[0089] More in particular, the invention provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues R-2 to V-87; V-3 to V-87; D-4 to V-87; T-5 to V-87; D-6 to V-87; E-7 to V-87; D-8 to V-87; R-9 to V-87; Y-10 to V-87; P-11 to V-87; Q-12 to V-87; K-13 to V-87; L-14 to V-87; A-15 to V-87; F-16 to V-87; A-17 to V-87; E-18 to V-87; C-19 to V-87; L-20 to V-87; C-21 to V-87; R-22 to V-87; G-23 to V-87; C-24 to V-87; I-25 to V-87; D-26 to V-87; A-27 to V-87; R-28 to V-87; T-29 to V-87; G-30 to V-87; R-31 to V-87; E-32 to V-87; T-33 to V-87; A-34 to V-87; A-35 to V-87; L-36 to V-87; N-37 to V-87; S-38 to V-87; V-39 to V-87; R-40 to V-87; L-41 to V-87; L-42 to V-87; Q-43 to V-87; S-44 to V-87; L-45 to V-87; L-46 to V-87; V-47 to V-87; L-48 to V-87; R-49 to V-87; R-50 to V-87; R-51 to V-87; P-52 to V-87; C-53 to V-87; S-54 to V-87; R-55 to V-87; D-56 to V-87; G-57 to V-87; S-58 to V-87; G-59 to V-87; L-60 to V-87; P-61 to V-87; T-62 to V-87; P-63 to V-87; G-64 to V-87; A-65 to V-87; F-66 to V-87; A-67 to V-87; F-68 to V-87; H-69 to V-87; T-70 to V-87; E-71 to V-87; F-72 to V-87; I-73 to V-87; H-74 to V-87; V-75 to V-87; P-76 to V-87; V-77 to V-87; G-78 to V-87; C-79 to V-87; T-80 to V-87; C-81 to V-87; and V-82 to V-87 of SEQ ID NO:2. Polypeptides encoded by these polynucleotides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-21 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0090] Further, the invention provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues S-2 to P-160; A-3 to P-160; R-4 to P-160; A-5 to P-160; R-6 to P-160; A-7 to P-160; V-8 to P-160; L-9 to P-160; S-10 to P-160; A-11 to P-160; F-12 to P-160; H-13 to P-160; H-14 to P-160; T-15 to P-160; L-16 to P-160; Q-17 to P-160; L-18 to P-160; G-19 to P-160; P-20 to P-160; R-21 to P-160; E-22 to P-160; Q-23 to P-160; A-24 to P-160; R-25 to P-160; N-26 to P-160; A-27 to P-160; S-28 to P-160; C-29 to P-160; P-30 to P-160; A-31 to P-160; G-32 to P-160; G-33 to P-160; R-34 to P-160; P-35 to P-160; A-36 to P-160; D-37 to P-160; R-38 to P-160; R-39 to P-160; F-40 to P-160; R-41 to P-160; P-42 to P-160; P-43 to P-160; T-44 to P-160; N-45 to P-160; L-46 to P-160; R-47 to P-160; S-48 to P-160; V-49 to P-160; S-50 to P-160; P-51 to P-160; W-52 to P-160; A-53 to P-160; Y-54 to P-160; R-55 to P-160; I-56 to P-160; S-57 to P-160; Y-58 to P-160; D-59 to P-160; P-60 to P-160; A-61 to P-160; R-62 to P-160; Y-63 to P-160; P-64 to P-160; R-65 to P-160; Y-66 to P-160; L-67 to P-160; P-68 to P-160; E-69 to P-160; A-70 to P-160; Y-71 to P-160; C-72 to P-160; L-73 to P-160; C-74 to P-160; R-75 to P-160; G-76 to P-160; C-77 to P-160; L-78 to P-160; T-79 to P-160; G-80 to P-160; L-81 to P-160; F-82 to P-160; G-83 to P-160; E-84 to P-160; E-85 to P-160; D-86 to P-160; V-87 to P-160; R-88 to P-160; F-89 to P-160; R-90 to P-160; S-91 to P-160; A-92 to P-160; P-93 to P-160; V-94 to P-160; Y-95 to P-160; M-96 to P-160; P-97 to P-160; T-98 to P-160; V-99 to P-160; V-100 to P-160; L-101 to P-160; R-102 to P-160; R-103 to P-160; T-104 to P-160; P-105 to P-160; A-106 to P-160; C-107 to P-160; A-108 to P-160; G-109 to P-160; G-110 to P-160; R-111 to P-160; S-112 to P-160; V-113 to P-160; Y-114 to P-160; T-115 to P-160; E-116 to P-160; A-117 to P-160; Y-118 to P-160; V-19 to P-160; T-120 to P-160; I-121 to P-160; P-122 to P-160; V-123 to P-160; G-124 to P-160; C-125 to P-160; T-126 to P-160; C-127 to P-160; V-128 to P-160; P-129 to P-160; E-130 to P-160; P-131 to P-160; E-132 to P-160; K-133 to P-160; D-134 to P-160; A-135 to P-160; D-136 to P-160; S-137 to P-160; I-138 to P-160; N-139 to P-160; S-140 to P-160; S-141 to P-160; I-142 to P-160; D-143 to P-160; K-144 to P-160; Q-145 to P-160; G-146 to P-160; A-147 to P-160; K-148 to P-160; L-149 to P-160; L-150 to P-160; L-151 to P-160; G-152 to P-160; P-153 to P-160; N-154 to P-160; and D-155 to P-160 of SEQ ID NO:4. Polypeptides encoded by these polynucleotides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-22 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0091] Also as mentioned above, even if deletion of one or more amino acids from the C-terminus of a protein results in modification of loss of one or more biological functions of the protein, other biological activities may still be retained. Thus, the ability of the shortened protein to induce and/or bind to antibodies which recognize the complete or mature IL-21 and IL-22 proteins generally will be retained when less than the majority of the residues of the complete or mature IL-21 and IL-22 proteins are removed from the C-terminus. Whether a particular polypeptide lacking C-terminal residues of a complete protein retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art.

[0092] Accordingly, the present invention further provides polypeptides having one or more residues removed from the carboxy terminus of the amino acid sequence of the IL-21 polypeptide shown in SEQ ID NO:2, up to the aspartic acid residue at position 6 of SEQ ID NO:2, and polynucleotides encoding such polypeptides. In addition, the present invention further provides polypeptides having one or more residues removed from the carboxy terminus of the amino acid sequence of the IL-22 polypeptide shown in SEQ ID NO:4, up to the arginine residues at position 6 of SEQ ID NO:4. In particular, the present invention provides polypeptides having the amino acid sequence of residues 1-m³ of the amino acid sequence in SEQ ID NO:2, where m³ is any integer in the range of 6 to 87, and residue 5 is the position of the first residue from the C-terminus of the complete IL-21 polypeptide (shown in SEQ ID NO:2) believed to be required for immunogenic activity of the IL-21 protein. In addition, the present invention also provides polypeptides having the amino acid sequence of residues 1-m⁴ of the amino acid sequence in SEQ ID NO:4, where m⁴ is any integer in the range of 6 to 160, and residue 5 is the position of the first residue from the C-terminus of the complete W-22 polypeptide (shown in SEQ ID NO:4) believed to be required for immunogenic activity of the IL-22 protein.

[0093] More in particular, the invention provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues A-1 to S-86; A-1 to R-85; A-1 to P-84; A-1 to L-83; A-1 to V-82; A-1 to C-81; A-1 to T-80; A-1 to C-79; A-1 to G-78; A-1 to V-77; A-1 to P-76; A-1 to V-75; A-1 to H-74; A-1 to 1-73; A-1 to F-72; A-1 to E-71; A-1 to T-70; A-1 to H-69; A-1 to F-68; A-1 to A-67; A-1 to F-66;

[0094] A-1 to A-65; A-1 to G-64; A-1 to P-63; A-1 to T-62; A-1 to P-61; A-1 to L-60; A-1 to G-59; A-1 to S-58; A-1 to G-57; A-1 to D-56; A-1 to R-55; A-1 to S-54; A-1 to C-53; A-i to P-52; A-1 to R-51; A-1 to R-50; A-1 to R-49; A-1 to L-48; A-1 to V-47; A-1 to L-46; A-1 to L-45; A-1 to S-44; A-1 to Q-43; A-1 to L-42; A-1 to L-41; A-1 to R-40; A-1 to V-39; A-1 to S-38; A-1 to N-37; A-1 to L-36; A-1 to A-35; A-1 to A-34; A-1 to T-33; A-1 to E-32; A-1 to R-31; A-1 to G-30; A-1 to T-29; A-1 to R-28; A-1 to A-27; A-1 to D-26; A-1 to I-25; A-1 to C-24; A-1 to G-23; A-1 to R-22; A-1 to C-21; A-1 to L-20; A-1 to C-19; A-1 to E-18; A-1 to A-17; A-1 to F-16; A-1 to A-15; A-1 to L-14; A-1 to K-13; A-1 to Q-12; A-1 to P-11; A-1 to Y-10; A-1 to R-9; A-1 to D-8; A-1 to E-7; and A-1 to D-6 of SEQ ID NO:2. Polynucleotides encoding these polypeptides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-21 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0095] Moreover, the invention also provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues N-1to G-159; N-1 to A-158; N-1 to P-157; N-1 to A-156; N-1 to D-155; N-1 to N-154; N-1 to P-153;N-1 to G-152; N-i to L-151; N-1 to L-150; N-1 to L-149; N-1 to K-148; N-1 to A-147; N-1 to G-146; N-1 to Q-145; N-1 to K-144; N-1 to D-143; N-1to I-142; N-1to S-141; N-1 to S-140; N-1 to N-139; N-1 to 1-138; N-1 to S-137; N-1 to D-136; N-1 to A-135; N-1 to D-134; N-1 to K-133; N-1 to E-132; N-1 to P-131; N-1 to E-130; N-1 to P-129; N-1to V-128; N-1 to C-127; N-1 to T-126; N-1 to C-125; N-1to G-124; N-1 to V-123; N-1to P-122; N-1 to I-121; N-1to T-120; N-1to V-119; N-1 to Y-118; N-1 to A-117; N-1 to E-116; N-1 to T-115; N-1to Y-114; N-1 to V-113; N-1 to S-112; N-1to R-111; N-1to G-110; N-1to G-109; N-1to A-108; N-1 to C-107; N-1 to A-106; N-1 to P-105; N-1 to T-104; N-1 to R-103; N-1 to R-102; N-1 to L-101; N-1 to V-100; N-1 to V-99; N-1to T-98; N-1 to P-97; N-1to M-96; N-1to Y-95; N-1to V-94; N-1to P-93; N-1 to A-92; N-1 to S-91; N-1 to R-90; N-1 to F-89; N-1 to R-88; N-1 to V-87; N-1 to D-86; N-1 to E-85; N-1 to E-84; N-1to G-83; N-1to F-82; N-1 to L-81; N-1 to G-80; N-1 to T-79; N-1 to L-78; N-1 to C-77; N-1 to G-76; N-1 to R-75; N-1 to C-74; N-1 to L-73; N-1 to C-72; N-1 to Y-71; N-1to A-70; N-1 to E-69; N-1to P-68; N-1 to L-67; N-1to Y-66; N-1to R-65; N-1 to P-64; N-1to Y-63; N-1 to R-62; N-1to A-61; N-1 to P-60; N-1to D-59; N-1 to Y-58; N-1 to S-57; N-1 to 1-56; N-1 to R-55; N-1 to Y-54; N-1 to A-53; N-1 to W-52; N-1to P-51; N-1 to S-50; N-1 to V-49; N-1 to S-48; N-1to R-47; N-1to L-46; N-1to N-45; N-1to T-44; N-1to P-43; N-1to P-42; N-1to R-41; N-1to F-40; N-1to R-39; N-1 to R-38; N-1 to D-37; N-1 to A-36; N-1 to P-35; N-1 to R-34; N-1 to G-33; N-1 to G-32; N-1to A-31; N-1to P-30; N-1 to C-29; N-1to S-28; N-1 to A-27; N-1to N-26; N-1 to R-25; N-1 to A-24; N-1 to Q-23; N-1 to E-22; N-1 to R-21; N-1to P-20; N-1 to G-19; N-1 to L-18; N-1 to Q-17; N-1 to L-16; N-1 to T-15; N-1 to H-14; N-1 to H-13; N-1 to F-12; N-1 to A-11; N-1 to S-10; N-1 to L-9; N-1 to V-8; N-1 to A-7; and N-1 to R-6 of SEQ ID NO:4. Polypeptides encoded by these polynucleotides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-22 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0096] The invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini of IL-21, which may be described generally as having residues n³-m³ of SEQ ID NO:2, where n³ and m³ are integers as described above. Likewise, the invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini of IL-22, which may be described generally as having residues n⁴-m⁴ of SEQ ID NO:4, where n⁴ and m⁴ are integers as described above.

[0097] Moreover, any polypeptide having one or more amino acids deleted from both the amino and the carboxyl termini of IL-22, described specifically as having residues n⁴-m⁴ of SEQ ID NO:4 (where n⁴ and m⁴ are integers as described above) may be excluded from the invention. In particular, any polypeptide having one or more amino acids deleted from both the amino and the carboxyl termini of IL-22 and which is defined by residues n⁴-m⁴ of SEQ ID NO:4, where n is equal to 21, 22, 23, 24 or 25 and m⁴ is equal to 271, 272, 273, 274, 275 or 276 may be excluded from the invention.

[0098] Also as mentioned above, even if deletion of one or more amino acids from the N-terminus of a protein results in modification or loss of one or more biological functions of the protein, other biological activities may still be retained. Thus, the ability of the shortened protein to induce and/or bind to antibodies which recognize the full-length or mature IL-21 polypeptides generally will be retained when less than the majority of the residues of the full-length or mature IL-21 polypeptides are removed from the N-terminus. Whether a particular polypeptide lacking N-terminal residues of a complete or full-length polypeptide retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art.

[0099] Accordingly, the present invention further provides polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of the IL-21 polypeptide shown in SEQ ID NO:29, up to the valine residue at position number 192, and polynucleotides encoding such polypeptides. In particular, the present invention provides polypeptides comprising the amino acid sequence of residues n⁵-197 of SEQ ID NO:29, where n⁵ is an integer in the range of 1 to 192, and 193 is the position of the first residue from the N-terminus of the full-length IL-21 polypeptide (shown in SEQ ID NO:29) believed to be required for immunogenic activity of the IL-21 protein.

[0100] More in particular, the invention provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues T-2 to V-197; L-3 to V-197; L-4 to V-197; P-5 to V-197; G-6 to V-197; L-7 to V-197; L-8 to V-197; F-9 to V-197; L-10 to V-197; T-11 to V-197; W-12 to V-197; L-13 to V-197; H-14 to V-197; T-15 to V-197; C-16 to V-197; L-17 to V-197; A-18 to V-197; H-19 to V-197; H-20 to V-197; D-21 to V-197; P-22 to V-197; S-23 to V-197; L-24 to V-197; R-25 to V-197; G-26 to V-197 ; H-27 to V-197 ; P-28 to V-197 ; H-29 to V-197 ; S-30 to V-197 ; H-31 to V-197; G-32 to V-197; T-33 to V-197; P-34 to V-197; H-35 to V-197; C-36 to V-197; Y-37 to V-197; S-38 to V-197; A-39 to V-197; E-40 to V-197; E-41 to V-197; L-42 to V-197; P-43 to V-197; L-44 to V-197; G-45 to V-197; Q-46 to V-197; A-47 to V-197; P-48 to V-197; P-49 to V-197; H-50 to V-197; L-51 to V-197; L-52 to V-197; A-53 to V-197; R-54 to V-197; G-55 to V-197; A-56 to V-197; K-57 to V-197; W-58 to V-197; G-59 to V-197; Q-60 to V-197; A-61 to V-197; L-62 to V-197; P-63 to V-197; V-64 to V-197; A-65 to V-197; L-66 to V-197; V-67 to V-197; S-68 to V-197; S-69 to V-197; L-70 to V-197; E-71 to V-197; A-72 to V-197; A-73 to V-197; S-74 to V-197; H-75 to V-197; R-76 to V-197; G-77 to V-197; R-78 to V-197; H-79 to V-197; E-80 to V-197; R-81 to V-197; P-82 to V-197; S-83 to V-197; A-84 to V-197; T-85 to V-197; T-86 to V-197; Q-87 to V-197; C-88 to V-197; P-89 to V-197; V-90 to V-197; L-91 to V-197; R-92 to V-197; P-93 to V-197; E-94 to V-197; E-95 to V-197; V-96 to V-197; L-97 to V-197; E-98 to V-197; A-99 to V-197; D-100 to V-197; T-101 to V-197; H-102 to V-197; Q-103 to V-197; R-104 to V-197; S-105 to V-197; I-106 to V-197; S-107 to V-197; P-108 to V-197; W-109 to V-197; R-110 to V-197; Y-111 to V-197; R-112 to V-197; V-113 to V-197; D-114 to V-197; T-115 to V-197; D-116 to V-197; E-117 to V-197; D-118 to V-197; R-119 to V-197; Y-120 to V-197; P-121 to V-197; Q-122 to V-197; K-123 to V-197; L-124 to V-197; A-125 to V-197; F-126 to V-197; A-127 to V-197; E-128 to V-197; C-129 to V-197; L-130 to V-197; C-131 to V-197; R-132 to V-197; G-133 to V-197; C-134 to V-197; I-135 to V-197; D-136 to V-197; A-137 to V-197; R-138 to V-197; T-139 to V-197; G-140 to V-197; R-141 to V-197; E-142 to V-197; T-143 to V-197; A-144 to V-197; A-145 to V-197; L-146 to V-197; N-147 to V-197; S-148 to V-197; V-149 to V-197; R-150 to V-197; L-151 to V-197; L-152 to V-197; Q-153 to V-197; S-154 to V-197; L-155 to V-197; L-156 to V-197; V-157 to V-197; L-158 to V-197; R-159 to V-197; R-160 to V-197; R-161 to V-197; P-162 to V-197; C-163 to V-197; S-164 to V-197; R-165 to V-197; D-166 to V-197; G-167 to V-197; S-168 to V-197; G-169 to V-197; L-170 to V-197; P-171 to V-197; T-172 to V-197; P-173 to V-197; G-174 to V-197; A-175 to V-197; F-176 to V-197; A-177 to V-197; F-178 to V-197; H-179 to V-197; T-180 to V-197; E-181 to V-197; F-182 to V-197; I-183 to V-197; H-184 to V-197; V-185 to V-197; P-186 to V-197; V-187 to V-197; G-188 to V-197; C-189 to V-197; T-190 to V-197; C-191 to V-197; and V-192 to V-197 of SEQ ID NO:29. Polypeptides encoded by these polynucleotides also are provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-21 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0101] Also as mentioned above, even if deletion of one or more amino acids from the C-terminus of a protein results in modification of loss of one or more biological functions of the protein, other biological activities may still be retained. Thus, the ability of the shortened polypeptide to induce and/or bind to antibodies which recognize the full-length or mature IL-21polypeptide generally will be retained when less than the majority of the residues of the full-length or mature IL-21 polypeptides are removed from the C-terminus. Whether a particular polypeptide lacking C-terminal residues of a complete protein retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art.

[0102] Accordingly, the present invention further provides polypeptides having one or more residues removed from the carboxy terminus of the amino acid sequence of the IL-21 polypeptide shown in SEQ ID NO:29, up to the glycine residue at position 6 of SEQ ID NO:29, and polynucleotides encoding such polypeptides. In particular, the present invention provides polypeptides having the amino acid sequence of residues 1-m⁵ of the amino acid sequence in SEQ ID NO:29, where m⁵ is any integer in the range of 6 to 196, and residue 5 is the position of the first residue from the C-terminus of the full-length IL-21 polypeptide (shown in SEQ ID NO:29) believed to be required for immunogenic activity of the IL-21 protein.

[0103] More in particular, the invention provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues M-1 to S-196; M-1 to R-195; M-1 to P-194; M-1 to L-193; M-1 to V-192; M-1 to C-191; M-1 to T-190; M-1 to C-189; M-1 to G-188; M-1 to V-187; M-1 to P-186; M-1 to V-185; M-1 to H-184; M-1 to 1-183; M-1 to F-182; M-1 to E-181; M-1 to T-180; M-1 to H-179; M-1 to F-178; M-1 to A-177; M-1 to F-176; M-1 to A-175; M-1 to G-174; M-1 to P-173; M-1 to T-172; M-1 to P-171; M-1 to L-170; M-1 to G-169; M-1 to S-168; M-1 to G-167; M-1 to D-166; M-1 to R-165; M-1 to S-164; M-1 to C-163; M-1 to P-162; M-1 to R-161; M-1 to R-160; M-1 to R-159; M-1 to L-158; M-1 to V-157; M-1 to L-156; M-1 to L-155; M-1 to S-154; M-1 to Q-153; M-1 to L-152; M-1 to L-151; M-1 to R-150; M-1 to V-149; M-1 to S-148; M-1 to N-147; M-1 to L-146; M-1 to A-145; M-1 to A-144; M-1 to T-143; M-1 to E-142; M-1 to R-141; M-1 to G-140; M-1 to T-139; M-1 to R-138; M-1 to A-137; M-1 to D-136; M-1 to I-135; M-1 to C-134; M-1 to G-133; M-1 to R-132; M-1 to C-131; M-1 to L-130; M-1 to C-129; M-1 to E-128; M-1 to A-127; M-1 to F-126; M-1 to A-125; M-1 to L-124; M-1 to K-123; M-1 to Q-122; M-1 to P-121; M-1 to Y-120; M-1 to R-119; M-1 to D-118; M-1 to E-117; M-1 to D-116; M-1 to T-115; M-1 to D-114; M-1 to V-113; M-1 to R-112; M-1 to Y-111; M-1 to R-110; M-1 to W-109; M-1 to P-108; M-1 to S-107; M-1 to I-106; M-1 to S-105; M-1 to R-104; M-1 to Q-103; M-1 to H-102; M-1 to T-101; M-1 to D-100; M-1 to A-99; M-1 to E-98; M-1 to L-97; M-1 to V-96; M-1 to E-95; M-1 to E-94; M-1 to P-93; M-1 to R-92; M-1 to L-91; M-1 to V-90; M-1 to P-89; M-1 to C-88; M-1 to Q-87; M-1 to T-86; M-1 to T-85; M-1 to A-84; M-1 to S-83; M-1 to P-82; M-1 to R-81; M-1 to E-80; M-1 to H-79; M-1 to R-78; M-I to G-77; M-1 to R-76; M-1 to H-75; M-I to S-74; M-1 to A-73; M-1 to A-72; M-1 to E-71; M-1 to L-70; M-1 to S-69; M-1 to S-68; M-1 to V-67; M-1 to L-66; M-1 to A-65; M-1 to V-64; M-1 to P-63; M-1 to L-62; M-1 to A-61; M-1 to Q-60; M-1 to G-59; M-1 to W-58; M-1 to K-57; M-1 to A-56; M-1 to G-55; M-I to R-54; M-1 to A-53; M-1 to L-52; M-1 to L-51; M-1 to H-50; M-I to P-49; M-1 to P-48; M-1 to A-47; M-1 to Q-46; M-1 to G-45; M-l to L-44; M-1 to P-43; M-1 to L-42; M-1 to E-41; M-I to E-40; M-I to A-39; M-1 to S-38; M-1 to Y-37; M-1 to C-36; M-1 to H-35; M-l to P-34; M-1 to T-33; M-1 to G-32; M-1 to H-31; M-1 to S-30; M-1 to H-29; M-1 to P-28; M-1 to H-27; M-1 to G-26; M-1 to R-25; M-1 to L-24; M-1 to S-23; M-1 to P-22; M-1 to D-21; M-1 to H-20; M-1 to H-19; M-1 to A-18; M-1 to L-17; M-1 to C-16; M-1 to T-15; M-1 to H-14; M-1 to L-13; M-1 to W-12; M-1 to T-11; M-1 to L-10; M-I to F-9; M-I to L-8; M-1 to L-7; and M-I to G-6 of SEQ ID NO:29. Polypeptides encoded by these polynucleotides also are provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-22 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0104] The invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini of IL-21, which may be described generally as having residues n⁵-m⁵ of SEQ ID NO:29, where n⁵ and m⁵ are integers as described above. Polynucleotides encoding such polypeptides are also provided.

[0105] Moreover, any polypeptide having one or more amino acids deleted from both the amino and the carboxyl termini of IL-21, described specifically as having residues n⁵-m⁵ of SEQ ID NO:29 (where n⁵ and m⁵ are integers as described above) may be excluded from the invention.

[0106] Also as mentioned above, even if deletion of one or more amino acids from the N-terminus of a protein results in modification or loss of one or more biological functions of the protein, other biological activities may still be retained. Thus, the ability of the shortened protein to induce and/or bind to antibodies which recognize the full-length, partial-length or mature IL-22 polypeptides generally will be retained when less than the majority of the residues of the full-length, partial-length or mature IL-22 polypeptides are removed from the N-terminus. Whether a particular polypeptide lacking N-terminal residues of a complete or full-length polypeptide retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art.

[0107] Accordingly, the present invention further provides polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of the IL-22 polypeptide shown in SEQ ID NO:32, up to the aspartic acid residue at position number 168, and polynucleotides encoding such polypeptides. In particular, the present invention provides polypeptides comprising the amino acid sequence of residues n⁶-173 of SEQ ID NO:32, where n⁶is an integer in the range of 1 to 168, and 168 is the position of the first residue from the N-terminus of the IL-22 polypeptide (shown in SEQ ID NO:32) believed to be required for immunogenic activity of the IL-22 protein.

[0108] More in particular, the invention provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues C-2 to P-173; A-3 to P-173; D-4 to P-173; R-5 to P-173; P-6 to P-173; E-7 to P-173; E-8 to P-173; L-9 to P-173; L-10 to P-173; E-11 to P-173; Q-12 to P-173; L-13 to P-173; Y-14 to P-173; G-15 to P-173; R-16 to P-173; L-17 to P-173; A-18 to P-173; A-19 to P-173; G-20 to P-173; V-21 to P-173; L-22 to P-173; S-23 to P-173; A-24 to P-173; F-25 to P-173; H-26 to P-173; H-27 to P-173; T-28 to P-173; L-29 to P-173; Q-30 to P-173; L-31 to P-173; G-32 to P-173; P-33 to P-173; R-34 to P-173; E-35 to P-173; Q-36 to P-173; A-37 to P-173; R-38 to P-173; N-39 to P-173; A-40 to P-173; S-41 to P-173; C-42 to P-173; P-43 to P-173; A-44 to P-173; G-45 to P-173; G-46 to P-173; R-47 to P-173; P-48 to P-173; A-49 to P-173; D-50 to P-173; R-51 to P-173; R-52 to P-173; F-53 to P-173; R-54 to P-173; P-55 to P-173; P-56 to P-173; T-57 to P-173; N-58 to P-173; L-59 to P-173; R-60 to P-173; S-61 to P-173; V-62 to P-173; S-63 to P-173; P-64 to P-173; W-65 to P-173; A-66 to P-173; Y-67 to P-173; R-68 to P-173; 1-69 to P-173; S-70 to P-173; Y-71 to P-173; D-72 to P-173; P-73 to P-173; A-74 to P-173; R-75 to P-173; Y-76 to P-173; P-77 to P-173; R-78 to P-173; Y-79 to P-173; L-80 to P-173; P-81 to P-173; E-82 to P-173; A-83 to P-173; Y-84 to P-173; C-85 to P-173; L-86 to P-173; C-87 to P-173; R-88 to P-173; G-89 to P-173; C-90 to P-173; L-91 to P-173; T-92 to P-173; G-93 to P-173; L-94 to P-173; F-95 to P-173; G-96 to P-173; E-97 to P-173; E-98 to P-173; D-99 to P-173; V-100 to P-173; R-101 to P-173; F-102 to P-173; R-103 to P-173; S-104 to P-173; A-105 to P-173; P-106 to P-173; V-107 to P-173; Y-108 to P-173; M-109 to P-173; P-110 to P-173; T-111 to P-173; V-112 to P-173; V-113 to P-173; L-114 to P-173; R-115 to P-173; R-116 to P-173; T-117 to P-173; P-118 to P-173; A-119 to P-173; C-120 to P-173; A-121 to P-173; G-122 to P-173; G-123 to P-173; R-124 to P-173; S-125 to P-173; V-126 to P-173; Y-127 to P-173; T-128 to P-173; E-129 to P-173; A-130 to P-173; Y-131 to P-173; V-132 to P-173; T-133 to P-173; I-134 to P-173; P-135 to P-173; V-136 to P-173; G-137 to P-173; C-138 to P-173; T-139 to P-173; C-140 to P-173; V-141 to P-173; P-142 to P-173; E-143 to P-173; P-144 to P-173; E-1345 to P-173; K-146 to P-173; D-147 to P-173; A-148 to P-173; D-149 to P-173; S-150 to P-173; I-151 to P-173; N-152 to P-173; S-153 to P-173; S-154 to P-173; I-155 to P-173; D-156 to P-173; K-157 to P-173; Q-158 to P-173; G-159 to P-173; A-160 to P-173; K-161 to P-173; L-162 to P-173; L-163 to P-173; L-164 to P-173; G-165 to P-173; P-166 to P-173; N-167 to P-173; and D-168 to P-173 of SEQ ID NO:32. Polypeptides encoded by these polynucleotides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-21 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0109] Also as mentioned above, even if deletion of one or more amino acids from the C-terminus of a protein results in modification of loss of one or more biological functions of the protein, other biological activities may still be retained. Thus, the ability of the shortened polypeptide to induce and/or bind to antibodies which recognize the full-length, partial-length or mature IL-22 polypeptide generally will be retained when less than the majority of the residues of the full-length, partial-length or mature IL-22 polypeptides are removed from the C-terminus. Whether a particular polypeptide lacking C-terminal residues of a complete protein retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art.

[0110] Accordingly, the present invention further provides polypeptides having one or more residues removed from the carboxy terminus of the amino acid sequence of the IL-22 polypeptide shown in SEQ ID NO:32, up to the proline residue at position 6 of SEQ ID NO:32, and polynucleotides encoding such polypeptides. In particular, the present invention provides polypeptides having the amino acid sequence of residues 1-m⁶ of the amino acid sequence in SEQ ID NO:32, where m⁶ is any integer in the range of 6 to 173, and residue 6 is the position of the first residue from the C-terminus of the IL-22 polypeptide (shown in SEQ ID NO:32) believed to be required for immunogenic activity of the IL-22 protein.

[0111] More in particular, the invention provides polynucleotides encoding polypeptides comprising, or alternatively consisting of, the amino acid sequence of residues G-1 to G-172; G-1 to A-171; G-1 to P-170; G-1 to A-169; G-1 to D-168; G-1 to N-167; G-1 to P-166; G-1 to G-165; G-1 to L-164; G-1 to L-163; G-1 to L-162; G-1 to K-161; G-1 to A-160; G-1 to G-159; G-1 to Q-158; G-1 to K-157; G-1 to D-156; G-1 to I-155; G-1 to S-154; G-1 to S-153; G-1 to N-152; G-1 to I-151; G-1 to S-150; G-1 to D-149; G-1 to A-148; G-1 to D-147; G-1 to K-146; G-1 to E-145; G-1 to P-144; G-1 to E-143; G-1 to P-142; G-1 to V-141; G-1 to C-140; G-1 to T-139; G-1 to C-138; G-1 to G-137; G-1 to V-136; G-1 to P-135; G-1 to I-134; G-1 to T-133; G-1 to V-132; G-1 to Y-131; G-1 to A-130; G-l to E-129; G-1 to T-128; G-1 to Y-127; G-1 to V-126; G-1 to S-125; G-1 to R-124; G-1 to G-123; G-1 to G-122; G-1 to A-121; G-1 to C-120; G-1 to A-119; G-1 to P-118; G-1 to T-117; G-1 to R-116; G-1 to R-115; G-1 to L-114; G-1 to V-113; G-1 to V-112; G-1 to T-111; G-1 to P-110; G-1 to M-109; G-1 to Y-108; G-1 to V-107; G-1 to P-106; G-1 to A-105; G-1 to S-104; G-1 to R-103; G-1 to F-102; G-1 to R-101; G-1 to V-100; G-1 to D-99; G-1 to E-98; G-1 to E-97; G-1 to G-96; G-1 to F-95; G-1 to L-94; G-1 to G-93; G-1 to T-92; G-1 to L-91; G-1 to C-90; G-1 to G-89; G-1 to R-88; G-1 to C-87; G-1 to L-86; G-1 to C-85; G-1 to Y-84; G-1 to A-83; G-1 to E-82; G-1 to P-81; G-1 to L-80; G-1 to Y-79; G-1 to R-78; G-1 to P-77; G-1 to Y-76; G-1 to R-75; G-1 to A-74; G-1 to P-73; G-1 to D-72; G-1 to Y-71; G-1 to S-70; G-1 to 1-69; G-1 to R-68; G-1 to Y-67; G-1 to A-66; G-1 to W-65; G-1 to P-64; G-1 to S-63; G-1 to V-62; G-1 to S-61; G-1 to R-60; G-1 to L-59; G-1 to N-58; G-1 to T-57; G-1 to P-56; G-1 to P-55; G-1 to R-54; G-1 to F-53; G-1 to R-52; G-1 to R-51; G-1 to D-50; G-1 to A-49; G-1 to P-48; G-1 to R-47; G-1 to G-46; G-1 to G-45; G-1 to A-44; G-1 to P-43; G-1 to C-42; G-1 to S-41; G-1 to A-40; G-1 to N-39; G-1 to R-38; G-1 to A-37; G-1 to Q-36; G-1 to E-35; G-1 to R-34; G-1 to P-33; G-1 to G-32; G-1 to L-31; G-1 to Q-30; G-1 to L-29; G-1 to T-28; G-1 to H-27; G-1 to H-26; G-1 to F-25; G-1 to A-24; G-1 to S-23; G-1 to L-22; G-1 to V-21; G-1 to G-20; G-1 to A-19; G-1 to A-18; G-1 to L-17; G-1 to R-16; G-1 to G-15; G-1 to Y-14; G-1 to L-13; G-1 to Q-12; G-1 to E-11; G-1 to L-10; G-1 to L-9; G-1 to E-8; G-1 to E-7; and G-1 to P-6 of SEQ ID NO:32. Polypeptides encoded by these polynucleotides are also provided. The present application is also directed to nucleic acid molecules comprising, or alternatively, consisting of, a polynucleotide sequence at least 90%, 95%, 96%, 97%, 98% or 99% identical to the polynucleotide sequence encoding the IL-22 polypeptides described above. The present invention also encompasses the above polynucleotide sequences fused to a heterologous polynucleotide sequence.

[0112] The invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini of IL-22, which may be described generally as having residues n⁶-m⁶ of SEQ ID NO:32, where n⁶ and m⁶ are integers as described above. Polynucleotides encoding these polypeptides are alos provided.

[0113] Moreover, any polypeptide having one or more amino acids deleted from both the amino and the carboxyl termini of IL-22, described specifically as having residues n⁶-m⁶ of SEQ ID NO:32 (where n⁶ and m⁶ are integers as described above) may be excluded from the invention, as may polynucleotides encoding such polypeptides.

[0114] The invention further includes IL-21 and IL-22 polypeptide variants which show substantial biological activity. Such variants include deletions, insertions, inversions, repeats, and substitutions selected according to general rules known in the art so as have little effect on activity. For example, guidance concerning how to make phenotypically silent amino acid substitutions is provided by Bowie and colleagues (Science 247:1306-1310 (1990)), wherein the authors indicate that there are two main strategies for studying the tolerance of an amino acid sequence to change.

[0115] The first strategy exploits the tolerance of amino acid substitutions by natural selection during the process of evolution. By comparing amino acid sequences in different species, conserved amino acids can be identified. These conserved amino acids are likely important for protein function. In contrast, the amino acid positions where substitutions have been tolerated by natural selection indicates that these positions are not critical for protein function. Thus, positions tolerating amino acid substitution could be modified while still maintaining biological activity of the protein.

[0116] The second strategy uses genetic engineering to introduce amino acid changes at specific positions of a cloned gene to identify regions critical for protein function. For example, site directed mutagenesis or alanine-scanning mutagenesis (introduction of single alanine mutations at every residue in the molecule) can be used (Cunningham and Wells, Science 244:1081-1085 (1989)). The resulting mutant molecules can then be tested for biological activity.

[0117] As the authors state, these two strategies have revealed that proteins are surprisingly tolerant of amino acid substitutions. The authors further indicate which amino acid changes are likely to be permissive at certain amino acid positions in the protein. For example, most buried (within the tertiary structure of the protein) amino acid residues require nonpolar side chains, whereas few features of surface side chains are generally conserved. Moreover, tolerated conservative amino acid substitutions involve replacement of an aliphatic or hydrophobic amino acid with another aliphatic or hydrophobic amino acid such as Ala, Val, Leu or Ile; replacement of a hydroxyl residue with another hydroxyl residue such as Ser or Thr; replacement of an acidic residue with another acidic residue such as Asp or Glu; replacement of an amide residue with another amide residue such as Asn or Gln, replacement of a basic residue with another basic residue such as Lys, Arg, or His; replacement of an aromatic residue with another aromatic residue such as Phe, Tyr, or Trp, and replacement of a small-sized amino acid with another small-sized residue such as Ala, Ser, Thr, Met, or Gly.

[0118] Besides conservative amino acid substitution, variants of IL-21 and IL-22 include (i) substitutions with one or more of the non-conserved amino acid residues, where the substituted amino acid residues may or may not be one encoded by the genetic code, or (ii) substitution with one or more of amino acid residues having a substituent group, or (iii) fusion of the mature polypeptide with another compound, such as a compound to increase the stability and/or solubility of the polypeptide (for example, polyethylene glycol), or (iv) fusion of the polypeptide with additional amino acids, such as an IgG Fc fusion region peptide, or leader or secretory sequence, or a sequence facilitating purification. Such variant polypeptides are deemed to be within the scope of those skilled in the art from the teachings herein.

[0119] For example, IL-21 and IL-22 polypeptide variants containing amino acid substitutions of charged amino acids with other charged or neutral amino acids may produce proteins with improved characteristics, such as less aggregation. Aggregation of pharmaceutical formulations both reduces activity and increases clearance due to the aggregate's immunogenic activity (Pinckard, et al., Clin. Exp. Immunol. 2:331-340 (1967); Robbins, et al., Diabetes 36:838-845 (1987); Cleland, et al., Crit. Rev. Ther. Drug Carrier Systems 10:307-377 (1993)).

[0120] Polynucleotide and Polypeptide Fragments

[0121] The invention provides nucleic acid molecules having nucleotide sequences related to extensive portions of SEQ ID NO:3 and SEQ ID NO:31 which have been determined from the following related cDNA clones: HE2CD08R (SEQ ID NO:24); HAGBX04R (SEQ ID NO:25); HCEBA24FB (SEQ ID NO:26); and HCELE59R (SEQ ID NO:27). Furthermore, the invention provides nucleic acid molecules having nucleotide sequences related to extensive portions of SEQ ID NO:28 which has been determined from a related cDNA clone designated HTGED19RB (SEQ ID NO:30). Such polynucleotides (i.e., SEQ ID NOs:24, 25, 26, and 30) may preferably be excluded from the present invention.

[0122] In the present invention, a polynucleotide fragment refers to a short polynucleotide having a nucleic acid sequence contained in the deposited clones or shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:28 or SEQ ID NO:31. The short nucleotide fragments are preferably at least about 15 nt, and more preferably at least about 20 nt, still more preferably at least about 30 nt, and even more preferably, at least about 40 nt in length. A fragment at least 20 nt in length, for example, is intended to include 20 or more contiguous bases from the cDNA sequence contained in the deposited clones or the nucleotide sequences shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:28 or SEQ ID NO:31. These nucleotide fragments are useful as diagnostic probes and primers as discussed herein. Of course, larger fragments (e.g., 50, 150, 500, 600, 2000 nucleotides) are preferred.

[0123] Moreover, representative examples of IL-21 polynucleotide fragments include, for example, fragments having a sequence from about nucleotide number 1-50, 51-100, 101-150, 151-200, 201-250, 251-300, 301-350, 351-400, 401-450, 451-500, 501-550, 551-600, 651-700, or 701 to the end of SEQ ID NO:1 or the cDNA contained in the deposited clone. In addition, representative examples of IL-22 polynucleotide fragments include, for example, fragments having a sequence from about nucleotide number 1-50, 51-100, 101-150, 151-200, 201-250, 251-300, 301-350, 351-400, 401-450, 451-500, 501-550, 551-600, 601-650, 651-700, 701-750, 751-800, 801-850, 851-900, 901-950, 951-1000, 1001-1050, 1051-1100, 1101-1150, 1151-1200, 1201-1250, 1251-1300, 1301-1350, 1351-1400, 1401-1450, 1451-1500, 1551-1600, or 1601 to the end of SEQ ID NO:3 or the cDNA contained in the deposited clone. Moreover, representative examples of the full-length IL-21 polynucleotide fragments include, for example, fragments having a sequence from about nucleotide number 1-1025, 50-1025, 100-1025, 150-1025, 200-1025, 250-1025, 300-1025, 350-1025, 400-1025, 450-1025, 500-1025, 550-1025, 600-1025, 650-1025, 700-1025, 750-1025, 800-1025, 850-1025, 900-1025, 950-1025, 1000-1025, 1-1000, 50-1000, 100-1000, 150-1000, 200-1000, 250-1000, 300-1000, 350-1000, 400-1000, 450-1000, 500-1000, 550-1000, 600-1000, 650-1000, 700-1000, 750-1000, 800-1000, 850-1000, 900-1000, 950-1000, 1-950, 50-950, 100-950, 150-950, 200-950, 250-950, 300-950, 350-950, 400-950, 450-950, 500-950, 550-950, 600-950, 650-950, 700-950, 750-950, 800-950, 850-950, 900-950, 1-900, 50-900, 100-900, 150-900, 200-900, 250-900, 300-900, 350-900, 400-900, 450-900, 500-900, 550-900, 600-900, 650-900, 700-900, 750-900, 800-900, 850-900, 1-850, 50-850, 100-850, 150-850, 200-850, 250-850, 300-850, 350-850, 400-850, 450-850, 500-850, 550-850, 600-850, 650-850, 700-850, 750-850, 800-850, 1-800, 50-800, 100-800, 150-800, 200-800, 250-800, 300-800, 350-800, 400-800, 450-800, 500-800, 550-800, 600-800, 650-800, 700-800, 750-800, 1-750, 50-750, 100-750, 150-750, 200-750, 250-750, 300-750, 350-750, 400-750, 450-750, 500-750, 550-750, 600-750, 650-750, 700-750, 1-700, 50-700, 100-700, 150-700, 200-700, 250-700, 300-700, 350-700, 400-700, 450-700, 500-700, 550-700, 600-700, 650-700, 1-650, 50-650, 100-650, 150-650, 200-650, 250-650, 300-650, 350-650, 400-650, 450-650, 500-650, 550-650, 600-650, 1-600, 50-600, 100-600, 150-600, 200-600, 250-600, 300-600, 350-600, 400-600, 450-600, 500-600, 550-600, 1-550, 50-550, 100-550, 150-550, 200-550, 250-550, 300-550, 350-550, 400-550, 450-550, 500-550, 1-500, 50-500, 100-500, 150-500, 200-500, 250-500, 300-500, 350-500, 400-500, 450-500, 1-450, 50-450, 100-450, 150-450, 200-450, 250-450, 300-450, 350-450, 400-450, 1-400, 50-400, 100-400, 150-400, 200-400, 250-400, 300-400, 350-400, 1-350, 50-350, 100-350, 150-350, 200-350, 250-350, 300-350, 1-300, 50-300, 100-300, 150-300, 200-300, 250-300, 1-250, 50-250, 100-250, 150-250, 200-250, 1-200, 50-200, 100-200, 150-200, 1-150, 50-150, 100-150, 1-100, 50-100, and 1-50 of SEQ ID NO:28. In this context about includes the particularly recited ranges, larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini. Preferably, these fragments encode a polypeptide which has biological activity.

[0124] Further, the invention includes a polynucleotide comprising any portion of at least about 30 nucleotides, preferably at least about 50 nucleotides, of SEQ ID NO:1 from residue 1 to 650, 25 to 650, 50 to 650, 75 to 650, 100 to 650, 125 to 650, 150 to 650, 175 to 650, 200 to 650, 225 to 650, 250 to 650, 275 to 650, 300 to 650, 325 to 650, 350 to 650, 375 to 650, 400 to 650, 425 to 650, 500 to 650, 525 to 650, 550 to 650, 575 to 650, 600 to 650, 625 to 650, 1 to 600, 25 to 600, 50 to 600, 75 to 600, 100 to 600, 125 to 600, 150 to 600, 175 to 600, 200 to 600, 225 to 600, 250 to 600, 275 to 600, 300 to 600, 325 to 600, 350 to 600, 375 to 600, 400 to 600, 425 to 600, 500 to 600, 525 to 600, 550 to 600, 575 to 600, 1 to 550, 25 to 550, 50 to 550, 75 to 550, 100 to 550, 125 to 550, 150 to 550, 175 to 550, 200 to 550, 225 to 550, 250 to 550, 275 to 550, 300 to 550, 325 to 550, 350 to 550, 375 to 550, 400 to 550, 425 to 550, 500 to 550, 525 to 550, 1 to 500, 25 to 500, 50 to 500, 75 to 500, 100 to 500, 125 to 500, 150 to 500, 175 to 500, 200 to 500, 225 to 500, 250 to 500, 275 to 500, 300 to 500, 325 to 500, 350 to 500, 375 to 500, 400 to 500, 425 to 500, 450 to 500, 475 to 500, 1 to 450, 25 to 450, 50 to 450, 75 to 450, 100 to 450, 125 to 450, 150 to 450, 175 to 450, 200 to 450, 225 to 450, 250 to 450, 275 to 450, 300 to 450, 325 to 450, 350 to 450, 375 to 450, 400 to 450, 425 to 450, 1 to 400, 25 to 400, 50 to 400, 75 to 400, 100 to 400, 125 to 400, 150 to 400, 175 to 400, 200 to 400, 225 to 400, 250 to 400, 275 to 400, 300 to 400, 325 to 400, 350 to 400, 375 to 400, 1 to 350, 25 to 350, 50 to 350, 75 to 350, 100 to 350, 125 to 350, 150 to 350, 175 to 350, 200 to 350, 225 to 350, 250 to 350, 275 to 350, 300 to 350, 325 to 350, 1 to 300, 25 to 300, 50 to 300, 75 to 300, 100 to 300, 125 to 300, 150 to 300, 175 to 300, 200 to 300, 225 to 300, 250 to 300, 275 to 300, 1 to 250, 25 to 250, 50 to 250, 75 to 250, 100 to 250, 125 to 250, 150 to 250, 175 to 250, 200 to 250, 225 to 250, 1 to 200, 25 to 200, 50 to 200, 75 to 200, 100 to 200, 125 to 200, 150 to 200, 175 to 200, 1 to 150, 25 to 150, 50 to 150, 75 to 150, 100 to 150, 125 to 150, 1 to 100, 25 to 100, 50 to 100, 75 to 100, 1 to 50, and 25 to 50.

[0125] Moreover, the invention includes a polynucleotide comprising any portion of at least about 30 nucleotides, preferably at least about 50 nucleotides, of SEQ ID NO:3 from residue 300 to 850. More preferably, the invention includes a polynucleotide comprising nucleotide residues 50 to 850, 75 to 850, 100 to 850, 125 to 850, 150 to 850, 175 to 850, 200 to 850, 225 to 850, 250 to 850, 275 to 850, 300 to 850, 325 to 850, 350 to 850, 375 to 850, 400 to 850, 425 to 850, 450 to 850, 475 to 850, 500 to 850, 525 to 850, 550 to 850, 575 to 850, 600 to 850, 625 to 850, 650 to 850, 675 to 850, 700 to 850, 750 to 850, 775 to 850, 800 to 850, 50 to 800, 75 to 800, 100 to 800, 125 to 800, 150 to 800, 175 to 800, 200 to 800, 225 to 800, 250 to 800, 275 to 800, 300 to 800, 325 to 800, 350 to 800, 375 to 800, 400 to 800, 425 to 800, 450 to 800, 475 to 800, 500 to 800, 525 to 800, 550 to 800, 575 to 800, 600 to 800, 625 to 800, 650 to 800, 675 to 800, 700 to 800, 750 to 800, 50 to 750, 75 to 750, 100 to 750, 125 to 750, 150 to 750, 175 to 750, 200 to 750, 225 to 750, 250 to 750, 275 to 750, 300 to 750, 325 to 750, 350 to 750, 375 to 750, 400 to 750, 425 to 750, 450 to 750, 475 to 750, 500 to 750, 525 to 750, 550 to 750, 575 to 750, 600 to 750, 625 to 750, 650 to 750, 675 to 750, 700 to 750, 50 to 700, 75 to 700, 100 to 700, 125 to 700, 150 to 700, 175 to 700, 200 to 700, 225 to 700, 250 to 700, 275 to 700, 300 to 700, 325 to 700, 350 to 700, 375 to 700, 400 to 700, 425 to 700, 450 to 700, 475 to 700, 500 to 700, 525 to 700, 550 to 700, 575 to 700, 600 to 700, 625 to 700, 650 to 700, 50 to 650, 75 to 650, 100 to 650, 125 to 650, 150 to 650, 175 to 650, 200 to 650, 225 to 650, 250 to 650, 275 to 650, 300 to 650, 325 to 650, 350 to 650, 375 to 650, 400 to 650, 425 to 650, 450 to 650, 475 to 650, 500 to 650, 525 to 650, 550 to 650, 575 to 650, 600 to 650, 50 to 600, 75 to 600, 100 to 600, 125 to 600, 150 to 600, 175 to 600, 200 to 600, 225 to 600, 250 to 600, 275 to 600, 300 to 600, 325 to 600, 350 to 600, 375 to 600, 400 to 600, 425 to 600, 450 to 600, 475 to 600, 500 to 600, 525 to 600, 550 to 600, 50 to 550, 75 to 550, 100 to 550, 125 to 550, 150 to 550, 175 to 550, 200 to 550, 225 to 550, 250 to 550, 275 to 550, 300 to 550, 325 to 550, 350 to 550, 375 to 550, 400 to 550, 425 to 550, 450 to 550, 475 to 550, 500 to 550, 50 to 500, 75 to 500, 100 to 500, 125 to 500, 150 to 500, 175 to 500, 200 to 500, 225 to 500, 250 to 500, 275 to 500, 300 to 500, 325 to 500, 350 to 500, 375 to 500, 400 to 500, 425 to 500, 450 to 500, 50 to 450, 75 to 450, 100 to 450, 125 to 450, 150 to 450, 175 to 450, 200 to 450, 225 to 450, 250 to 450, 275 to 450, 300 to 450, 325 to 450, 350 to 450, 375 to 450, 400 to 450, 50 to 400, 75 to 400, 100 to 400, 125 to 400, 150 to 400, 175 to 400, 200 to 400, 225 to 400, 250 to 400, 275 to 400, 300 to 400, 325 to 400, 350 to 400, 50 to 350, 75 to 350, 100 to 350, 125 to 350, 150 to 350, 175 to 350, 200 to 350, 225 to 350, 250 to 350, 275 to 350, 300 to 350, 50 to 300, 75 to 300, 100 to 300, 125 to 300, 150 to 300, 175 to 300, 200 to 300, 225 to 300, and 250 to 300.

[0126] In the present invention, a polypeptide fragment refers to a short amino acid sequence contained in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:29, SEQ ID NO:32 or encoded by the cDNAs contained in the deposited clones. Protein fragments may be free-standing, or comprised within a larger polypeptide of which the fragment forms a part or region, most preferably as a single continuous region. Representative examples of polypeptide fragments of the partial IL-21 invention, include, for example, fragments from about amino acid number 1-20, 21-40, 41-60, 61-83 or to the end of the coding region. Moreover, polypeptide fragments of IL-21 can be about 10, 20, 30, 40, 50, 60, 70, or 80 amino acids in length. Representative examples of polypeptide fragments of the IL-22 invention, include, for example, fragments from about amino acid number 1-20, 21-40, 41-60, 61-80, 81-100, 100-120, 120-140, 140-160, or to the end of the coding region. Moreover, polypeptide fragments of IL-22 can be about 10, 20, 30, 40, 50, 60, 70, 80, 100, 120, 140, or 150 amino acids in length. Representative examples of polypeptide fragments of the full-length IL-21 of the invention, include, for example, fragments from about amino acid number 1-20, 21-40, 41-60, 61-80, 81-100, 100-120, 120-140, 140-160, 160-180, 180-200 or 180-to the end of the coding region. Moreover, polypeptide fragments of the full-length IL-21 can be about 10, 20, 30, 40, 50, 60, 70, 80, 100, 120, 140, 150, 160, 170, 180 or 190 amino acids in length. In this context about includes the particularly recited ranges, larger or smaller by several (5, 4, 3, 2, or 1) amino acids, at either extreme or at both extremes.

[0127] A further embodiment of the invention relates to a peptide or polypeptide which comprises the amino acid sequence of an IL-21 or EL-22 polypeptide having an amino acid sequence which contains at least one conservative amino acid substitution, but not more than 50 conservative amino acid substitutions, even more preferably, not more than 40 conservative amino acid substitutions, still more preferably, not more than 30 conservative amino acid substitutions, and still even more preferably, not more than 20 conservative amino acid substitutions. Of course, in order of ever-increasing preference, it is highly preferable for a peptide or polypeptide to have an amino acid sequence which comprises the amino acid sequence of an IL-21 or IL-22 polypeptide, which contains at least one, but not more than 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 conservative amino acid substitutions.

[0128] Preferred polypeptide fragments include the secreted IL-21 and IL-22 proteins as well as the mature forms. Further preferred polypeptide fragments include the secreted IL-21 and IL-22 proteins or the mature forms having a continuous series of deleted residues from the amino or the carboxy terminus, or both. For example, any number of amino acids, ranging from 1-60, can be deleted from the amino terminus of either the secreted or the mature form of the IL-21 and IL-22 polypeptides. Similarly, any number of amino acids, ranging from 1-30, can be deleted from the carboxy terminus of the secreted or the mature form of the IL-21 and IL-22 polypeptides. Furthermore, any combination of the above amino and carboxy terminus deletions are preferred. Similarly, polynucleotide fragments encoding these IL-21 and IL-22 polypeptide fragments are also preferred. Also preferred are IL-21 and IL-22 polypeptide and polynucleotide fragments .characterized by structural or functional domains, such as fragments that comprise alpha-helix and alpha-helix forming regions, beta-sheet and beta-sheet-forming regions, turn and turn-forming regions, coil and coil-forming regions, hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, flexible regions, surface-forming regions, substrate binding region, and high antigenic index regions. Polypeptide fragments of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:29 or SEQ ID NO:32 falling within conserved domains are specifically contemplated by the present invention (FIGS. 4, 5, 7, and 9). Moreover, polynucleotide fragments encoding these domains are also contemplated. , In additional embodiments, the polynucleotides of the invention encode functional attributes of IL-21 or IL-22. Preferred embodiments of the invention in this regard include fragments that comprise alpha-helix and alpha-helix forming regions (alpha-regions), beta-sheet and beta-sheet forming regions (beta-regions), turn and turn-forming regions (turn-regions), coil and coil-forming regions (coil-regions), hydrophilic regions, hydrophobic regions, alpha amphipathic regions, beta amphipathic regions, flexible regions, surface-forming regions and high antigenic index regions of IL-21 or IL-22.

[0129] The data representing the structural or functional attributes of IL-21 set forth in FIG. 7 and/or Table I, as described above, was generated using the various modules and algorithms of the DNA*STAR set on default parameters. The data representing the structural or functional attributes of IL-22 set forth in FIG. 5 and/or Table II, in FIG. 9 and/or Table III, as described above, was generated using the various modules and algorithms of the DNA*STAR set on default parameters. In a preferred embodiment, the data presented in columns VIII, IX, XIII, and XIV of Table I can be used to determine regions of L-21 which exhibit a high degree of potential for antigenicity. In an additional preferred embodiment, the data presented in columns VIII, IX, XIII, and XIV of Tables II and/or III can be used to determine regions of IL-22 which exhibit a high degree of potential for antigenicity. Regions of high antigenicity are determined from the data presented in columns VIII, IX, XIII, and/or IV by choosing values which represent regions of the polypeptide which are likely to be exposed on the surface of the polypeptide in an environment in which antigen recognition may occur in the process of initiation of an immune response.

[0130] Certain preferred regions in these regards are set out in FIG. 7, but may, as shown in Tables I, be represented or identified by using tabular representations of the data presented in FIG. 7. The DNA* STAR computer algorithm used to generate FIG. 7 (set on the original default parameters) was used to present the data in FIG. 7 in a tabular format (See Table I). The tabular format of the data in FIG. 7 may be used to easily determine specific boundaries of a preferred region.

[0131] Certain preferred regions in these regards are set out in FIGS. 5 and 8, but may, as shown in Tables II and III, respectively, be represented or identified by using tabular representations of the data presented in FIGS. 5 and 8, respectively. The DNA*STAR computer algorithm used to generate FIGS. 5 and 8 (set on the original default parameters) was used to present the data in FIGS. 5 and 8 in a tabular format (See Tables II and III, respectively). The tabular format of the data in FIGS. 5 and 8 may be used to easily determine specific boundaries of a preferred region.

[0132] The above-mentioned preferred regions set out in FIGS. 5, 7, and 9, and in Tables II, I, and III, respectively, include, but are not limited to, regions of the aforementioned types identified by analysis of the amino acid sequence set out in FIGS. 2A-B, 6A-B, and 8, respectively. As set out in FIG. 7 and in Table I, and in FIG. 5 and Table II, and in FIG. 8 and Table III, such preferred regions include Garnier-Robson alpha-regions, beta-regions, turn-regions, and coil-regions, Chou-Fasman alpha-regions, beta-regions, and coil-regions, Kyte-Doolittle hydrophilic regions and hydrophobic regions, Eisenberg alpha- and beta-amphipathic regions, Karplus-Schulz flexible regions, Emini surface-forming regions and Jameson-Wolf regions of high antigenic index.

TABLE I
Res	Position	I	II	III	IV	V	VI	VII	VIII	IX	X	XI	XII	XIII	XIV
Met	1	A	.	.	.	.	.	.	0.80	0.76	.	.	.	0.40	0.36
Thr	2	.	.	B	.	.	.	.	0.76	0.76	.	.	.	0.40	0.44
Leu	3	A	.	.	.	.	.	.	1.18	0.76	.	.	.	0.40	0.34
Leu	4	A	.	.	.	.	T	.	1.60	1.01	.	.	.	0.20	0.28
Pro	5	A	.	.	.	.	T	.	1.91	1.09	.	.	F	0.05	0.16
Gly	6	A	.	.	.	.	T	.	2.12	1.39	.	.	.	0.20	0.17
Leu	7	A	.	.	.	.	T	.	2.12	1.39	.	.	.	0.20	0.17
Leu	8	A	A	.	.	.	.	.	1.60	1.19	.	.	.	0.60	0.16
Phe	9	A	A	.	.	.	.	.	1.60	1.67	.	.	.	0.60	0.17
Leu	10	A	A	.	.	.	.	.	1.42	1.93	.	.	.	0.60	0.17
Thr	11	A	A	.	.	.	.	.	1.39	1.74	.	.	.	0.60	0.28
Trp	12	A	A	.	.	.	.	.	1.24	1.54	.	.	.	0.60	0.46
Leu	13	A	A	.	.	.	.	.	1.24	1.33	.	.	.	0.60	0.30
His	14	A	A	.	.	.	.	.	1.13	1.33	*	.	.	0.60	0.17
Thr	15	A	A	.	.	.	.	.	0.36	1.34	.	.	.	0.60	0.17
Cys	16	A	A	.	.	.	.	.	0.08	0.93	.	.	.	0.60	0.27
Leu	17	A	A	.	.	.	.	.	0.21	0.74	.	.	.	0.60	0.27
Ala	18	.	A	.	.	T	.	.	0.81	0.24	.	.	.	0.10	0.32
His	19	.	A	.	.	T	.	.	0.54	0.19	.	.	.	0.44	0.91
His	20	.	A	.	.	.	.	C	0.04	0.00	.	*	.	1.33	1.48
Asp	21	.	.	.	.	.	T	C	0.82	0.00	.	*	F	2.22	1.21
Pro	22	.	.	.	.	T	T	.	1.29	0.50	.	*	F	2.76	1.74
Ser	23	.	.	.	.	T	T	.	1.84	0.57	.	*	F	3.40	1.27
Leu	24	.	.	.	.	T	T	.	1.67	0.57	.	*	F	3.06	1.03
Arg	25	.	.	.	.	T	.	.	1.67	0.14	*	*	F	2.35	1.03
Gly	26	.	.	.	.	T	.	.	1.37	0.07	*	*	F	2.14	1.05
His	27	.	.	.	.	.	T	C	1.54	0.07	.	*	.	1.78	1.70
Pro	28	.	.	.	.	.	T	C	1.50	0.26	.	*	.	1.57	1.18
His	29	.	.	.	.	T	T	.	2.00	0.17	.	*	.	1.30	1.18
Ser	30	.	.	.	.	T	T	.	1.68	0.23	.	*	.	1.17	1.26
His	31	.	.	.	.	T	.	.	1.99	0.16	.	.	.	0.84	1.26
Gly	32	.	.	.	.	T	.	.	1.36	0.23	.	.	F	0.86	1.26
Thr	33	.	.	.	.	.	T	C	1.32	0.30	.	.	F	0.58	0.50
Pro	34	.	.	.	.	.	T	C	1.06	0.67	.	.	F	0.15	0.58
His	35	.	.	.	.	T	T	.	0.77	0.56	.	.	.	0.20	0.78
Cys	36	.	.	.	.	T	T	.	0.80	0.63	.	.	.	0.20	0.55
Tyr	37	.	A	.	.	T	.	C	1.14	0.14	.	.	.	0.10	0.61
Ser	38	.	A	.	.	.	.	C	0.64	0.29	.	.	.	0.50	0.78
Ala	39	A	A	.	.	.	.	.	0.64	0.10	.	.	.	0.45	1.20
Glu	40	A	A	.	.	.	.	.	0.13	0.24	.	.	F	0.60	1.19
Glu	41	A	A	.	.	.	.	.	0.19	0.31	.	.	.	0.30	0.73
Leu	42	A	.	.	.	.	T	.	0.43	0.27	.	.	.	0.70	0.72
Pro	43	A	.	.	.	.	T	.	0.14	0.37	.	.	.	0.70	0.72
Leu	44	.	.	.	.	T	T	.	0.52	0.13	.	.	.	0.50	0.42
Gly	45	.	.	.	.	T	T	.	0.31	0.56	.	.	F	0.35	0.78
Gln	46	A	.	.	.	.	.	.	0.28	0.30	.	.	F	0.05	0.78
Ala	47	.	.	.	.	.	.	C	0.28	0.37	*	.	F	0.40	1.29
Pro	48	.	.	.	.	.	T	C	0.32	0.37	*	.	F	0.60	1.08
Pro	49	A	.	.	.	.	T	.	0.10	0.63	*	*	F	0.05	0.51
His	50	A	.	.	.	.	T	.	0.36	0.73	*	*	.	0.20	0.51
Leu	51	A	.	.	.	.	T	.	0.01	0.23	*	*	.	0.10	0.65
Leu	52	A	A	.	.	.	.	.	0.01	0.23	*	*	.	0.30	0.42
Ala	53	A	A	.	.	.	.	.	0.27	0.30	*	.	.	0.30	0.31
Arg	54	A	A	.	.	.	.	.	0.19	0.20	*	.	.	0.30	0.75
Gly	55	A	A	.	.	.	.	.	0.12	0.03	*	.	F	0.15	0.95
Ala	56	A	A	.	.	.	.	.	0.69	0.23	*	.	F	0.45	0.93
Lys	57	.	A	.	.	T	.	.	0.91	0.33	*	.	F	0.85	0.83
Trp	58	.	A	.	.	T	.	.	0.69	0.17	*	.	F	0.25	0.84
Gly	59	.	A	.	.	.	.	C	0.37	0.43	*	.	F	0.25	0.69
Gln	60	A	A	.	.	.	.	.	0.14	0.36	*	.	.	0.30	0.53
Ala	61	.	A	.	.	.	.	C	0.14	1.00	*	.	.	0.40	0.38
Leu	62	.	A	B	.	.	.	.	1.00	0.59	*	.	.	0.60	0.38
Pro	63	.	A	B	.	.	.	.	1.57	0.84	.	.	.	0.60	0.18
Val	64	A	A	.	.	.	.	.	1.52	1.09	.	.	.	0.60	0.13
Ala	65	A	A	.	.	.	.	.	1.82	0.97	.	.	.	0.60	0.22
Leu	66	A	A	.	.	.	.	.	2.04	0.67	.	.	.	0.60	0.19
Val	67	A	A	.	.	.	.	.	1.23	0.93	.	.	.	0.60	0.21
Ser	68	A	A	.	.	.	.	.	1.61	0.29	.	.	.	0.30	0.36
Ser	69	A	A	.	.	.	.	.	1.34	0.29	.	.	.	0.30	0.44
Leu	70	A	A	.	.	.	.	.	1.06	0.10	*	.	.	0.30	0.60
Glu	71	A	A	.	.	.	.	.	0.28	0.16	*	.	.	0.30	0.60
Ala	72	A	A	.	.	.	.	.	0.69	0.04	*	.	.	0.30	0.61
Ala	73	A	A	.	.	.	.	.	0.64	0.43	*	*	.	0.79	1.45
Ser	74	A	.	.	.	.	.	.	1.06	0.69	*	*	.	1.48	0.83
His	75	A	.	.	.	.	T	.	1.83	0.69	*	*	.	2.17	1.60
Arg	76	A	.	.	.	.	T	.	1.83	0.69	.	*	F	2.66	2.16
Gly	77	.	.	.	.	T	T	.	2.53	1.19	.	*	F	3.40	2.79
Arg	78	.	.	.	.	T	T	.	2.91	1.57	.	*	F	3.06	4.02
His	79	.	.	.	.	.	.	C	2.91	1.64	.	*	F	2.66	3.17
Glu	80	.	.	.	.	.	.	C	2.36	1.26	.	*	F	2.66	4.29
Arg	81	.	.	.	.	.	T	C	1.93	1.19	.	*	F	2.86	2.21
Pro	82	.	.	.	.	T	T	.	1.97	0.70	.	.	F	3.06	2.35
Ser	83	.	.	.	.	T	T	.	1.86	0.71	.	*	F	3.40	1.96
Ala	84	.	.	.	.	T	T	.	1.22	0.31	.	*	F	2.76	1.73
Thr	85	.	.	.	B	T	.	.	1.01	0.26	.	*	F	1.27	0.60
Thr	86	.	.	.	B	T	.	.	0.04	0.26	.	*	F	0.93	0.69
Gln	87	.	.	.	B	T	.	.	0.56	0.51	.	.	F	0.29	0.51
Cys	88	.	.	B	B	.	.	.	0.14	0.70	*	.	.	0.60	0.29
Pro	89	.	.	B	B	.	.	.	0.23	0.21	.	*	.	0.30	0.39
Val	90	.	.	.	B	.	.	C	0.54	0.16	.	.	.	0.10	0.35
Leu	91	.	A	.	.	.	.	C	0.86	0.24	.	.	.	0.65	1.14
Arg	92	.	A	.	.	.	.	C	0.00	0.81	*	.	F	1.10	1.27
Pro	93	A	A	.	.	.	.	.	0.14	0.60	*	*	F	0.90	1.27
Glu	94	A	A	.	.	.	.	.	0.07	0.56	*	*	F	0.90	1.27
Glu	95	A	A	.	.	.	.	.	0.33	1.24	*	*	F	0.90	1.13
Val	96	A	A	.	.	.	.	.	1.14	0.74	.	*	.	0.60	0.74
Leu	97	A	A	.	.	.	.	.	0.72	1.17	*	*	.	0.60	0.71
Glu	98	A	A	.	.	.	.	.	0.90	0.69	.	.	.	0.60	0.59
Ala	99	A	A	.	.	.	.	.	0.90	0.19	.	*	F	0.60	1.08
Asp	100	A	.	.	.	.	T	.	1.01	0.43	.	*	F	1.00	2.28
Thr	101	A	.	.	.	.	T	.	1.57	1.11	*	*	F	1.30	2.58
His	102	A	.	.	.	.	T	.	1.49	0.73	*	*	F	1.30	3.42
Gln	103	.	.	.	.	T	T	.	1.19	0.54	*	.	F	1.91	1.43
Arg	104	.	.	.	B	T	.	.	1.57	0.16	*	.	F	1.42	1.33
Ser	105	.	.	.	B	T	.	.	1.28	0.21	*	*	F	1.63	1.51
Ile	106	.	.	.	B	.	.	C	1.70	0.20	*	*	F	0.89	0.92
Ser	107	.	.	.	.	.	T	C	1.49	0.20	*	*	F	2.10	0.92
Pro	108	.	.	.	.	T	T	.	1.60	0.56	*	*	F	1.34	1.07
Trp	109	.	.	.	.	T	T	.	0.63	0.17	*	*	.	1.28	3.00
Arg	110	.	.	.	.	.	T	C	0.93	0.13	.	*	.	0.87	1.66
Tyr	111	.	.	.	B	T	.	.	1.51	0.26	.	*	.	1.40	1.80
Arg	112	.	.	.	B	T	.	.	1.81	0.20	.	*	.	1.53	2.46
Val	113	.	.	.	B	.	.	C	2.02	1.11	.	*	.	1.97	2.10
Asp	114	.	.	.	.	T	T	.	2.31	1.11	.	*	F	3.06	2.32
Thr	115	.	.	.	.	T	T	.	2.31	1.87	.	*	F	3.40	1.98
Asp	116	.	.	.	.	T	T	.	2.31	1.87	*	*	F	3.06	5.23
Glu	117	.	.	.	.	I	T	.	1.99	1.76	*	*	F	2.72	4.90
Asp	118	.	.	.	.	T	T	.	2.84	1.33	*	.	F	2.38	5.25
Arg	119	A	.	.	.	.	T	.	2.89	1.41	*	*	F	1.64	5.45
Tyr	120	A	.	.	.	.	T	.	2.39	1.41	*	.	F	1.30	6.29
Pro	121	A	.	.	.	.	T	.	1.80	0.73	*	*	F	1.30	3.11
Gln	122	A	A	.	.	.	.	.	1.10	0.23	*	*	F	0.60	1.60
Lys	123	A	A	.	.	.	.	.	0.51	0.56	*	*	F	0.45	0.89
Leu	124	A	A	.	.	.	.	.	0.40	0.30	*	*	.	0.30	0.58
Ala	125	A	A	.	.	.	.	.	0.02	0.13	.	.	.	0.30	0.58
Phe	126	A	A	.	.	.	.	.	0.62	0.04	.	.	.	0.30	0.16
Ala	127	A	A	.	.	.	.	.	1.29	0.73	*	.	.	0.60	0.16
Glu	128	A	A	.	.	.	.	.	1.22	0.61	*	*	.	0.60	0.08
Cys	129	A	A	.	.	.	.	.	0.76	0.11	*	.	.	0.30	0.19
Leu	130	A	A	.	.	.	.	.	0.83	0.24	*	*	.	0.30	0.18
Cys	131	.	.	.	.	T	T	.	1.02	0.17	*	*	.	1.10	0.06
Arg	132	.	.	.	.	T	T	.	0.43	0.51	*	*	.	0.20	0.07
Gly	133	.	.	.	.	T	T	.	1.02	0.06	*	*	.	1.10	0.15
Cys	134	.	.	.	.	T	T	.	0.24	0.24	*	*	.	1.40	0.28
Ile	135	A	.	.	.	.	.	.	0.26	0.81	*	*	.	1.40	0.28
Asp	136	.	.	.	.	T	.	.	0.58	0.33	.	*	.	1.80	0.41
Ala	137	.	.	.	.	T	.	.	0.58	0.33	.	*	.	2.10	0.76
Arg	138	.	.	.	.	.	T	C	0.92	0.90	*	*	F	3.00	2.12
Thr	139	.	.	.	.	.	T	C	1.28	1.59	*	*	F	2.70	2.20
Gly	140	.	.	.	.	.	T	C	1.58	1.10	*	*	F	2.40	3.14
Arg	141	A	.	.	.	.	T	.	0.99	1.10	*	.	F	1.90	1.62
Glu	142	A	A	.	.	.	.	.	0.77	0.60	*	*	F	1.20	1.13
Thr	143	A	A	.	.	.	.	.	0.66	0.40	*	.	F	0.45	0.94
Ala	144	A	A	.	.	.	.	.	0.67	0.43	.	*	.	0.30	0.78
Ala	145	A	A	.	.	.	.	.	0.16	0.04	.	*	.	0.30	0.60
Leu	146	A	.	.	B	.	.	.	0.16	0.60	.	*	.	0.60	0.31
Asn	147	A	.	.	B	.	.	.	0.66	0.11	*	.	.	0.30	0.60
Ser	148	A	.	.	B	.	.	.	1.16	0.30	*	.	.	0.30	0.49
Val	149	A	.	.	B	.	.	.	0.57	0.49	*	.	.	0.60	0.49
Arg	150	A	.	.	B	.	.	.	0.28	0.20	*	.	.	0.30	0.53
Leu	151	A	.	.	B	.	.	.	0.28	0.19	*	.	.	0.30	0.53
Leu	152	A	.	.	B	.	.	.	1.09	0.49	*	*	.	0.60	0.58
Gln	153	A	.	.	B	.	.	.	1.64	0.53	*	*	.	0.60	0.25
Ser	154	A	.	.	B	.	.	.	1.60	1.17	.	*	.	0.60	0.22
Leu	155	.	.	B	B	.	.	.	1.60	1.17	*	.	.	0.60	0.22
Leu	156	.	.	B	B	.	.	.	0.68	0.49	*	*	.	0.60	0.25
Val	157	.	.	B	B	.	.	.	0.24	0.09	*	*	.	0.30	0.37
Leu	158	.	.	B	B	.	.	.	0.03	0.30	*	.	.	0.30	0.87
Arg	159	.	.	.	B	T	.	.	0.33	0.56	.	.	F	1.30	1.63
Arg	160	.	.	.	B	T	.	.	0.18	0.67	.	*	F	1.30	1.18
Arg	161	.	.	.	B	.	.	C	1.10	0.93	.	*	F	1.10	1.91
Pro	162	.	.	.	.	T	.	.	1.96	1.61	.	*	F	1.84	1.91
Cys	163	.	.	.	.	T	.	.	2.42	1.61	.	*	F	2.18	1.63
Ser	164	.	.	.	.	T	T	.	2.01	1.19	.	*	F	2.57	0.82
Arg	165	.	.	.	.	T	T	.	1.56	0.80	*	.	F	2.91	0.71
Asp	166	.	.	.	.	T	T	.	0.63	0.80	*	*	F	3.40	1.32
Gly	167	.	.	.	.	T	T	.	0.63	0.69	*	.	F	2.91	0.81
Ser	168	.	.	.	.	T	.	.	0.99	0.64	*	.	F	2.37	0.64
Gly	169	.	.	.	.	.	.	C	1.08	0.16	*	.	F	1.53	0.55
Leu	170	.	.	.	.	.	.	C	0.62	0.27	*	.	F	0.59	0.87
Pro	171	.	.	.	.	.	.	C	0.03	0.27	.	.	F	0.25	0.64
Thr	172	.	.	.	.	.	T	C	0.32	0.39	.	.	F	0.45	0.65
Pro	173	.	.	.	.	.	T	C	0.61	0.74	.	.	F	0.15	0.68
Gly	174	.	.	.	.	.	T	C	0.97	0.56	.	.	F	0.15	0.45
Ala	175	A	.	.	.	.	T	.	0.19	0.91	.	.	.	0.20	0.27
Phe	176	A	A	.	.	.	.	.	0.29	0.93	.	.	.	0.60	0.24
Ala	177	A	A	.	.	.	.	.	0.02	0.99	.	*	.	0.60	0.34
Phe	178	A	A	.	.	.	.	.	0.47	0.56	.	.	.	0.60	0.59
His	179	A	A	.	.	.	.	.	1.01	0.84	.	.	.	0.60	0.59
Thr	180	A	.	.	B	.	.	.	0.46	0.74	.	.	.	0.60	0.41
Glu	181	A	.	.	B	.	.	.	0.61	0.74	.	.	.	0.60	0.64
Phe	182	A	.	.	B	.	.	.	0.23	0.60	.	.	.	0.60	0.35
Ile	183	.	.	.	B	T	.	.	0.39	0.53	.	.	.	0.20	0.38
His	184	.	.	.	B	T	.	.	0.70	0.69	.	.	.	0.20	0.16
Val	185	.	.	.	B	.	.	C	1.06	1.11	.	.	.	0.40	0.18
Pro	186	.	.	.	.	T	T	.	1.37	0.90	.	.	.	0.20	0.14
Val	187	.	.	.	.	T	T	.	1.33	0.70	.	.	.	0.20	0.15
Gly	188	.	.	.	.	T	T	.	1.30	0.77	.	*	.	0.20	0.11
Cys	189	.	.	.	.	T	T	.	2.08	0.77	.	.	.	0.20	0.05
Thr	190	.	.	B	B	.	.	.	1.43	1.03	.	*	.	0.60	0.06
Cys	191	.	.	B	B	.	.	.	1.11	0.81	.	.	.	0.60	0.09
Val	192	.	.	B	B	.	.	.	0.56	0.39	*	.	.	0.30	0.33
Leu	193	.	.	B	.	.	T	.	1.07	0.20	*	.	.	0.28	0.31
Pro	194	.	.	B	.	.	T	.	0.79	0.36	*	.	F	0.61	0.42
Arg	195	.	.	.	.	T	T	.	0.87	0.21	*	.	.	1.04	0.73
Ser	196	.	.	.	.	T	T	.	0.59	0.00	*	.	.	1.97	1.13
Val	197	.	.	.	.	T	.	.	0.12	0.26	*	.	.	1.80	0.93

[0133]

TABLE II
Res	Position	I	II	III	IV	V	VI	VII	VIII	IX	X	XI	XII	XIII
Asn	1	.	.	.	.	.	.	C	0.58	.	*	.	0.85	1.60
Ser	2	.	A	.	.	.	.	C	1.08	.	*	.	0.65	1.26
Ala	3	.	A	B	.	.	.	.	0.88	.	*	.	0.75	1.93
Arg	4	.	A	B	.	.	.	.	0.41	.	*	.	0.75	1.22
Ala	5	.	A	B	.	.	.	.	0.01	.	*	.	0.30	0.67
Arg	6	.	A	B	.	.	.	.	0.31	.	*	.	0.30	0.55
Ala	7	.	A	B	.	.	.	.	0.60	.	*	.	0.30	0.38
Val	8	.	A	B	.	.	.	.	0.71	.	*	.	0.30	0.38
Leu	9	.	A	B	.	.	.	.	0.86	*	*	.	0.60	0.17
Ser	10	.	A	B	.	.	.	.	0.30	*	*	.	0.60	0.22
Ala	11	.	A	B	.	.	.	.	0.72	*	.	.	0.60	0.41
Phe	12	.	A	B	.	.	.	.	0.94	*	.	.	0.60	0.72
His	13	.	A	B	.	.	.	.	0.09	*	.	.	0.60	0.44
His	14	.	A	B	.	.	.	.	0.09	*	.	.	0.60	0.76
Thr	15	.	A	B	.	.	.	.	0.13	*	.	.	0.60	0.72
Leu	16	.	A	.	.	.	.	C	0.24	*	*	.	0.10	0.52
Gln	17	.	A	.	.	T	.	.	1.06	*	*	.	0.40	0.60
Leu	18	.	A	.	.	.	.	C	1.09	.	*	.	0.80	0.81
Gly	19	.	.	.	.	.	T	C	1.12	.	*	F	2.40	1.70
Pro	20	.	.	.	.	.	T	C	0.84	*	*	F	3.00	1.70
Arg	21	.	.	.	.	.	T	C	1.77	*	*	F	2.70	2.08
Glu	22	.	.	B	.	.	T	.	1.77	*	*	F	2.20	4.12
Gln	23	.	.	B	.	.	.	.	1.99	*	*	F	1.70	4.28
Ala	24	.	.	.	.	T	.	.	2.03	*	*	F	1.80	2.21
Arg	25	.	.	.	.	T	.	.	1.58	*	*	F	1.50	1.71
Asn	26	.	.	.	.	T	.	.	1.26	*	*	F	1.05	0.53
Ala	27	.	.	.	.	T	.	.	0.67	*	.	.	0.90	0.81
Ser	28	.	.	B	.	.	.	.	0.32	.	.	.	0.78	0.42
Cys	29	.	.	B	.	.	T	.	0.57	.	*	.	0.66	0.26
Pro	30	.	.	.	.	T	T	.	0.57	.	*	.	1.34	0.25
Ala	31	.	.	.	.	T	T	.	0.36	.	*	F	2.37	0.37
Gly	32	.	.	.	.	T	T	.	0.36	.	.	F	2.80	1.06
Gly	33	.	.	.	.	.	.	C	0.66	*	*	F	1.97	0.69
Arg	34	.	.	B	.	.	.	.	1.43	*	.	F	1.94	1.15
Pro	35	.	.	B	.	.	T	.	1.76	*	.	F	1.86	2.27
Ala	36	.	.	B	.	.	T	.	1.64	*	*	F	1.58	4.49
Asp	37	.	.	B	.	.	T	.	2.10	*	*	F	1.30	1.99
Arg	38	.	.	B	.	.	T	.	2.23	*	*	F	1.30	2.52
Arg	39	.	.	B	.	.	.	.	1.91	*	*	F	1.10	3.85
Phe	40	.	.	B	.	.	.	.	1.81	*	*	F	1.44	3.57
Arg	41	.	.	B	.	.	.	.	2.40	*	*	F	1.78	2.63
Pro	42	.	.	.	.	.	T	C	1.59	.	*	F	2.22	2.16
Pro	43	.	.	.	.	T	T	.	1.59	.	*	F	2.16	2.05
Thr	44	.	.	.	.	T	T	.	1.18	.	*	F	3.40	2.05
Asn	45	.	.	.	.	.	T	C	1.02	*	*	F	2.56	1.78
Leu	46	.	.	B	B	.	.	.	0.61	*	*	F	0.87	0.85
Arg	47	.	.	B	B	.	.	.	0.61	*	.	F	1.13	0.79
Ser	48	.	.	B	B	.	.	.	0.53	*	.	F	0.79	0.76
Val	49	.	.	B	B	.	.	.	0.26	*	.	F	0.45	0.97
Ser	50	.	.	B	.	.	T	.	0.01	*	*	F	0.25	0.50
Pro	51	.	.	B	.	.	T	.	0.93	*	*	.	0.20	0.59
Trp	52	.	.	B	.	.	T	.	0.07	*	*	.	0.05	1.55
Ala	53	.	.	B	.	.	T	.	0.07	*	*	.	0.20	0.81
Tyr	54	.	.	B	B	.	.	.	0.54	*	*	.	0.60	0.70
Arg	55	.	.	B	B	.	.	.	0.84	.	*	.	0.45	1.05
Ile	56	.	.	B	B	.	.	.	0.84	*	*	.	0.13	1.73
Ser	57	.	.	B	.	.	.	.	0.54	*	*	.	0.61	1.71
Tyr	58	.	.	.	.	T	.	.	1.24	*	*	.	1.74	0.88
Asp	59	.	.	.	.	.	T	C	1.24	*	*	F	2.32	2.46
Pro	60	.	.	.	.	T	T	.	0.92	*	.	F	2.80	2.88
Ala	61	.	.	.	.	T	T	.	1.92	*	.	F	2.52	2.84
Arg	62	.	.	B	.	.	T	.	1.98	*	.	F	2.14	3.33
Tyr	63	.	.	B	.	.	T	.	1.41	*	.	.	1.41	3.37
Pro	64	.	.	B	.	.	T	.	1.20	*	.	.	0.53	2.75
Arg	65	.	.	.	.	T	T	.	1.41	*	.	.	0.65	2.17
Tyr	66	.	.	B	.	.	T	.	1.41	*	.	F	0.40	2.40
Leu	67	.	.	B	.	.	.	.	1.06	*	.	F	0.80	1.57
Pro	68	.	.	B	.	.	.	.	0.63	*	.	.	0.05	1.26
Glu	69	.	.	.	.	T	.	.	0.03	*	.	.	0.00	0.43
Ala	70	.	.	B	B	.	.	.	0.74	*	.	.	0.60	0.43
Tyr	71	.	.	B	B	.	.	.	0.39	*	.	.	0.60	0.15
Cys	72	.	.	B	B	.	.	.	0.08	*	.	.	0.30	0.17
Leu	73	.	.	B	B	.	.	.	0.38	.	*	.	0.60	0.16
Cys	74	.	.	B	.	.	T	.	1.19	.	*	.	0.20	0.06
Arg	75	.	.	B	.	.	T	.	0.91	*	*	.	0.20	0.09
Gly	76	.	.	B	.	.	T	.	1.01	*	.	.	0.20	0.15
Cys	77	.	.	B	.	.	T	.	1.16	.	*	.	0.10	0.28
Leu	78	.	.	B	B	.	.	.	1.04	.	.	.	0.30	0.12
Thr	79	.	.	B	B	.	.	.	0.72	.	*	.	0.60	0.10
Gly	80	.	.	.	B	.	.	C	0.83	.	*	.	0.40	0.19
Leu	81	.	.	.	B	.	.	C	0.49	.	.	.	0.40	0.40
Phe	82	.	.	B	B	.	.	.	0.18	.	.	F	0.45	0.48
Gly	83	.	.	.	B	.	.	C	0.13	.	*	F	0.95	0.81
Glu	84	.	A	B	.	.	.	.	0.56	.	*	F	0.45	0.73
Glu	85	.	A	B	.	.	.	.	0.20	.	*	F	0.90	1.65
Asp	86	.	A	B	B	.	.	.	1.12	.	*	F	0.90	1.45
Val	87	.	A	B	B	.	.	.	1.52	.	*	F	0.90	1.63
Arg	88	.	A	.	B	T	.	.	1.28	.	*	.	1.15	1.26
Phe	89	.	A	.	B	T	.	.	1.07	.	*	.	1.00	0.77
Arg	90	.	A	.	B	T	.	.	0.21	.	*	.	0.85	1.59
Ser	91	.	A	.	B	.	.	C	0.03	.	*	.	0.50	0.60
Ala	92	.	.	.	B	.	.	C	0.22	.	*	.	0.25	1.09
Pro	93	.	.	.	B	.	.	C	0.10	.	*	.	0.10	0.55
Val	94	.	.	.	B	T	.	.	0.29	*	.	.	0.20	0.64
Tyr	95	.	.	B	B	.	.	.	0.68	*	.	.	0.60	0.91
Met	96	.	.	B	B	.	.	.	1.23	.	.	.	0.60	0.44
Pro	97	.	.	B	B	.	.	.	1.46	.	*	.	0.60	0.44
Thr	98	.	.	B	B	.	.	.	1.13	*	.	.	0.60	0.23
Val	99	.	.	B	B	.	.	.	0.17	*	.	.	0.60	0.46
Val	100	.	.	B	B	.	.	.	0.23	.	.	.	0.30	0.58
Leu	101	.	.	B	B	.	.	.	0.16	.	.	.	0.30	0.58
Arg	102	.	.	B	B	.	.	.	0.22	.	.	F	0.60	1.20
Arg	103	.	.	B	B	.	.	.	0.58	.	.	F	0.60	1.63
Thr	104	.	.	B	B	.	.	.	0.31	.	.	F	0.60	1.06
Pro	105	.	.	B	B	.	.	.	0.20	*	.	F	1.00	0.55
Ala	106	.	.	B	.	.	.	.	0.67	.	*	.	1.00	0.28
Cys	107	.	.	B	.	.	T	.	0.67	.	.	.	0.85	0.19
Ala	108	.	.	.	.	T	T	.	0.26	*	*	.	2.10	0.24
Gly	109	.	.	.	.	T	T	.	0.29	*	.	F	2.50	0.32
Gly	110	.	.	.	.	T	T	.	0.32	*	.	F	2.25	0.44
Arg	111	.	.	B	B	.	.	.	0.04	*	.	F	0.60	0.69
Ser	112	.	.	B	B	.	.	.	0.62	*	.	F	0.35	1.00
Val	113	.	.	B	B	.	.	.	0.62	*	.	.	0.70	1.75
Tyr	114	.	.	B	.	.	.	.	0.72	.	.	.	0.50	0.90
Thr	115	.	.	B	.	.	.	.	0.21	.	.	.	0.25	1.05
Glu	116	.	.	B	B	.	.	.	0.21	.	*	.	0.45	1.05
Ala	117	.	.	B	B	.	.	.	0.80	.	*	.	0.60	0.97
Tyr	118	.	.	B	B	.	.	.	0.16	.	*	.	0.60	0.47
Val	119	.	.	B	B	.	.	.	0.77	.	*	.	0.60	0.42
Thr	120	.	.	B	B	.	.	.	0.80	.	*	.	0.60	0.31
Ile	121	.	.	B	B	.	.	.	1.47	.	*	.	0.60	0.20
Pro	122	.	.	B	.	.	T	.	1.19	.	*	.	0.20	0.14
Val	123	.	.	.	.	T	T	.	1.61	.	.	.	0.20	0.14
Gly	124	.	.	.	.	T	T	.	1.61	.	.	.	0.20	0.11
Cys	125	.	.	B	.	.	T	.	1.51	.	.	.	0.20	0.05
Thr	126	.	.	B	.	.	.	.	0.62	.	.	.	0.40	0.11
Cys	127	.	.	B	.	.	.	.	0.62	.	.	.	0.10	0.19
Val	128	.	.	B	.	.	T	.	0.23	.	.	.	0.40	0.55
Pro	129	.	.	B	.	.	T	.	0.62	.	.	F	1.45	0.65
Glu	130	.	.	B	.	.	T	.	1.29	*	.	F	2.20	2.44
Pro	131	.	.	B	.	.	T	.	1.01	*	.	F	2.50	5.49
Glu	132	.	.	.	.	T	.	.	1.68	*	.	F	3.00	3.59
Lys	133	A	.	.	.	.	.	.	2.23	*	.	F	2.30	3.46
Asp	134	A	.	.	.	.	T	.	1.56	*	.	F	2.20	3.00
Ala	135	A	.	.	.	.	T	.	1.56	*	.	F	1.90	1.21
Asp	136	A	.	.	.	.	T	.	1.47	*	.	F	1.45	0.98
Ser	137	.	.	B	.	.	T	.	1.17	*	.	F	1.15	0.78
Ile	138	.	.	B	.	.	.	.	0.23	*	.	F	0.80	1.04
Asn	139	.	.	B	.	.	T	.	0.23	*	.	F	0.85	0.44
Ser	140	.	.	B	.	.	T	.	0.87	*	.	F	1.16	0.54
Ser	141	.	.	B	.	.	T	.	0.87	*	.	F	1.62	1.55
Ile	142	.	.	B	.	.	T	.	0.82	.	*	F	2.23	1.67
Asp	143	.	.	B	.	.	T	.	1.12	*	*	F	2.54	1.23
Lys	144	.	.	.	.	T	T	.	1.17	*	.	F	3.10	0.93
Gln	145	.	.	B	.	.	T	.	0.66	*	.	F	2.54	2.65
Gly	146	.	.	B	.	.	T	.	0.14	*	.	F	2.23	1.31
Ala	147	.	A	B	.	.	.	.	0.22	*	.	F	1.07	0.54
Lys	148	.	A	B	.	.	.	.	0.12	.	.	F	0.16	0.26
Leu	149	.	A	B	.	.	.	.	0.38	*	.	.	0.60	0.26
Leu	150	.	A	B	.	.	.	.	0.38	.	.	.	0.60	0.39
Leu	151	.	A	B	.	.	.	.	0.03	.	.	.	0.06	0.32
Gly	152	.	.	B	.	.	T	.	0.03	.	.	F	0.73	0.64
Pro	153	.	.	.	.	.	T	C	0.29	.	.	F	1.17	0.78
Asn	154	.	.	.	.	T	T	.	0.07	.	.	F	2.36	1.47
Asp	155	.	.	.	.	.	T	C	0.40	.	.	F	2.40	1.50
Ala	156	.	.	.	.	.	.	C	1.00	.	.	F	1.81	0.96
Pro	157	.	.	.	.	.	T	C	0.96	.	.	F	1.77	0.92
Ala	158	.	.	.	.	.	T	C	0.78	.	.	.	1.38	0.71
Gly	159	.	.	.	.	.	T	C	0.39	.	.	.	0.54	0.90
Pro	160	.	.	B	.	.	T	.	0.00	.	.	.	0.10	0.74

[0134]

TABLE III
Res	Position	I	II	III	IV	V	VI	VII	VIII		IX	X	XI	XII	XIII
Gly	1	.	A	.	.	T	.	.	0.46	0.21	.	.	.	0.70	0.34
Cys	2	.	A	.	.	T	.	.	0.63	0.64	.	.	.	1.00	0.51
Ala	3	.	A	.	.	.	.	C	1.02	0.64	.	.	.	0.80	0.62
Asp	4	.	A	.	.	.	.	C	1.41	1.07	.	.	.	0.95	1.09
Arg	5	A	A	.	.	.	.	.	0.99	1.50	*	.	F	0.90	3.51
Pro	6	A	A	.	.	.	.	.	0.52	1.39	*	.	F	0.90	2.87
Glu	7	A	A	.	.	.	.	.	1.19	1.20	*	.	F	0.90	1.42
Glu	8	A	A	.	.	.	.	.	1.78	1.20	*	.	F	0.90	1.25
Leu	9	A	A	.	.	.	.	.	0.97	0.80	*	.	F	0.90	1.40
Leu	10	A	A	.	.	.	.	.	0.61	0.54	*	.	F	0.75	0.67
Glu	11	A	A	.	.	.	.	.	0.48	0.21	*	*	.	0.30	0.60
Gln	12	A	A	.	B	.	.	.	0.59	0.64	*	*	.	0.60	0.73
Leu	13	A	A	.	B	.	.	.	0.22	0.04	*	*	.	0.45	1.72
Tyr	14	A	A	.	B	.	.	.	0.00	0.04	*	*	.	0.30	0.82
Gly	15	A	A	.	.	.	.	.	0.22	0.46	*	*	.	0.60	0.48
Arg	16	A	A	.	.	.	.	.	0.12	0.56	*	*	.	0.60	0.59
Leu	17	A	A	.	.	.	.	.	0.98	0.30	*	*	.	0.30	0.37
Ala	18	A	A	.	B	.	.	.	0.98	0.19	*	*	.	0.30	0.28
Ala	19	A	A	.	B	.	.	.	1.03	0.44	*	*	.	0.60	0.12
Gly	20	A	A	.	B	.	.	.	1.28	0.83	*	*	.	0.60	0.19
Val	21	A	A	.	B	.	.	.	2.09	0.64	*	.	.	0.60	0.19
Leu	22	A	A	.	B	.	.	.	1.31	0.93	.	.	.	0.60	0.16
Ser	23	A	A	.	.	.	.	.	0.76	0.93	.	.	.	0.60	0.22
Ala	24	A	A	.	.	.	.	.	0.48	1.00	.	.	.	0.60	0.41
Phe	25	A	A	.	B	.	.	.	0.94	0.84	*	.	.	0.60	0.72
His	26	A	A	.	B	.	.	.	0.09	0.84	*	*	.	0.60	0.44
His	27	.	A	B	B	.	.	.	0.09	0.86	*	.	.	0.60	0.76
Thr	28	.	A	B	B	.	.	.	0.13	1.04	.	.	.	0.60	0.72
Leu	29	.	A	.	B	.	.	C	0.24	0.69	*	*	.	0.10	0.52
Gln	30	.	A	.	B	T	.	.	1.06	0.61	*	*	.	0.40	0.60
Leu	31	.	A	.	B	.	.	C	1.09	0.11	.	*	.	0.80	0.81
Gly	32	.	.	.	.	.	T	C	1.12	0.37	.	*	F	2.40	1.70
Pro	33	.	.	.	.	.	T	C	0.84	0.66	*	*	F	3.00	1.70
Arg	34	.	.	.	.	.	T	C	1.77	0.56	*	*	F	2.70	2.08
Glu	35	A	.	.	.	.	T	.	1.77	1.24	*	*	F	2.20	4.12
Gln	36	A	.	.	.	.	.	.	1.99	1.27	*	*	F	1.70	4.28
Ala	37	.	.	.	.	T	.	.	2.03	1.20	*	*	F	1.80	2.21
Arg	38	.	.	.	.	T	.	.	1.58	0.81	*	*	F	1.50	1.71
Asn	39	.	.	.	.	T	.	.	1.26	0.24	*	*	F	1.05	0.53
Ala	40	.	.	.	.	T	.	.	0.67	0.21	*	.	.	0.90	0.81
Ser	41	.	.	B	.	.	.	.	0.32	0.21	.	.	.	0.78	0.42
Cys	42	.	.	B	.	.	T	.	0.57	0.21	.	*	.	0.66	0.26
Pro	43	.	.	.	.	T	T	.	0.57	0.24	.	*	.	1.34	0.25
Ala	44	.	.	.	.	T	T	.	0.36	0.26	.	*	F	2.37	0.37
Gly	45	.	.	.	.	T	T	.	0.36	0.21	.	.	F	2.80	1.06
Gly	46	.	.	.	.	.	.	C	0.66	0.29	*	*	F	1.97	0.69
Arg	47	.	.	B	.	.	.	.	1.43	0.71	*	.	F	1.94	1.15
Pro	48	.	.	B	.	.	T	.	1.76	1.21	*	.	F	1.86	2.27
Ala	49	.	.	B	.	.	T	.	1.64	1.64	*	*	F	1.58	4.49
Asp	50	.	.	B	.	.	T	.	2.10	1.29	*	*	F	1.30	1.99
Arg	51	.	.	B	.	.	T	.	2.23	1.29	*	*	F	1.30	2.52
Arg	52	.	.	B	.	.	.	.	1.91	1.29	*	*	F	1.10	3.85
Phe	53	.	.	B	.	.	.	.	1.81	1.36	*	*	F	1.44	3.57
Arg	54	.	.	B	.	.	.	.	2.40	0.87	*	*	F	1.78	2.63
Pro	55	.	.	.	.	.	T	C	1.59	0.47	.	*	F	2.22	2.16
Pro	56	.	.	.	.	T	T	.	1.59	0.21	.	*	F	2.16	2.05
Thr	57	.	.	.	.	T	T	.	1.18	0.57	.	*	F	3.40	2.05
Asn	58	.	.	.	.	.	T	C	1.02	0.19	*	*	F	2.56	1.78
Leu	59	.	.	B	B	.	.	.	0.61	0.03	*	*	F	0.87	0.85
Arg	60	.	.	B	B	.	.	.	0.61	0.01	*	*	F	1.13	0.79
Ser	61	.	.	B	B	.	.	.	0.53	0.07	*	.	F	0.79	0.76
Val	62	.	.	B	B	.	.	.	0.26	0.44	*	.	F	0.45	0.97
Ser	63	.	.	B	.	.	T	.	0.01	0.26	*	*	F	0.25	0.50
Pro	64	.	.	B	.	.	T	.	0.93	1.01	*	*	.	0.20	0.59
Trp	65	.	.	B	.	.	T	.	0.07	0.63	*	*	.	0.05	1.55
Ala	66	.	.	B	.	.	T	.	0.07	0.67	*	*	.	0.20	0.81
Tyr	67	.	.	B	B	.	.	.	0.54	0.67	*	*	.	0.60	0.70
Arg	68	.	.	B	B	.	.	.	0.84	1.00	.	*	.	0.45	1.05
Ile	69	.	.	B	B	.	.	.	0.84	0.09	*	*	.	0.13	1.73
Ser	70	.	.	B	.	.	.	.	0.54	0.01	*	*	.	0.61	1.71
Tyr	71	.	.	.	.	T	.	.	1.24	0.24	*	*	.	1.74	0.88
Asp	72	.	.	.	.	.	T	C	1.24	0.24	*	*	F	2.32	2.46
Pro	73	.	.	.	.	T	T	.	0.92	0.17	*	*	F	2.80	2.88
Ala	74	.	.	.	.	T	T	.	1.92	0.13	*	.	F	2.52	2.84
Arg	75	.	.	B	.	.	T	.	1.98	0.89	*	.	F	2.14	3.33
Tyr	76	.	.	B	.	.	T	.	1.41	0.13	*	.	.	1.41	3.37
Pro	77	.	.	B	.	.	T	.	1.20	0.13	*	.	.	0.53	2.75
Arg	78	.	.	.	.	T	T	.	1.41	0.06	*	.	.	0.65	2.17
Tyr	79	.	.	B	.	.	T	.	1.41	0.06	*	.	F	0.40	2.40
Leu	80	.	.	B	.	.	.	.	1.06	0.20	*	.	F	0.80	1.57
Pro	81	.	.	B	.	.	.	.	0.63	0.13	*	.	.	0.05	1.26
Glu	82	.	.	.	.	T	.	.	0.03	0.70	*	.	.	0.00	0.43
Ala	83	.	.	B	B	.	.	.	0.74	0.63	*	.	.	0.60	0.43
Tyr	84	.	.	B	B	.	.	.	0.39	0.51	.	.	.	0.60	0.15
Cys	85	.	.	B	B	.	.	.	0.08	0.09	*	.	.	0.30	0.17
Leu	86	.	.	B	B	.	.	.	0.38	0.51	.	*	.	0.60	0.16
Cys	87	.	.	B	.	.	T	.	1.19	0.59	.	*	.	0.20	0.06
Arg	88	.	.	B	.	.	T	.	0.91	0.51	*	*	.	0.20	0.09
Gly	89	.	.	B	.	.	T	.	1.01	0.43	*	.	.	0.20	0.15
Cys	90	.	.	B	.	.	T	.	1.16	0.17	.	*	.	0.10	0.28
Leu	91	.	.	B	B	.	.	.	1.04	0.29	.	.	.	0.30	0.12
Thr	92	.	.	B	B	.	.	.	0.72	1.07	.	*	.	0.60	0.10
Gly	93	.	.	.	B	.	.	C	0.83	1.07	.	*	.	0.40	0.19
Leu	94	.	.	.	B	.	.	C	0.49	0.50	.	.	.	0.40	0.40
Phe	95	.	.	B	B	.	.	.	0.18	0.19	.	.	F	0.45	0.48
Gly	96	A	.	.	B	.	.	.	0.13	0.67	.	*	F	0.75	0.81
Glu	97	A	A	.	.	.	.	.	0.56	0.46	.	*	F	0.45	0.73
Glu	98	A	A	.	.	.	.	.	0.20	1.14	.	*	F	0.90	1.65
Asp	99	A	A	.	B	.	.	.	1.12	1.14	.	*	F	0.90	1.45
Val	100	A	A	.	B	.	.	.	1.52	1.57	.	*	F	0.90	1.63
Arg	101	A	A	.	B	.	.	.	1.28	1.19	.	*	.	0.75	1.26
Phe	102	A	A	.	B	.	.	.	1.07	0.69	.	*	.	0.60	0.77
Arg	103	A	A	.	B	.	.	.	0.21	0.26	.	*	.	0.45	1.59
Ser	104	.	A	.	B	.	.	C	0.03	0.26	.	*	.	0.50	0.60
Ala	105	.	.	.	B	.	.	C	0.22	0.50	.	*	.	0.25	1.09
Pro	106	.	.	.	B	.	.	C	0.10	0.33	.	*	.	0.10	0.55
Val	107	.	.	.	B	T	.	.	0.29	0.76	*	.	.	0.20	0.64
Tyr	108	.	.	B	B	.	.	.	0.68	0.86	*	.	.	0.60	0.91
Met	109	.	.	B	B	.	.	.	1.23	1.00	.	.	.	0.60	0.44
Pro	110	.	.	B	B	.	.	.	1.46	1.21	.	*	.	0.60	0.44
Thr	111	.	.	B	B	.	.	.	1.13	1.26	*	.	.	0.60	0.23
Val	112	.	.	B	B	.	.	.	0.17	0.50	*	.	.	0.60	0.46
Val	113	.	.	B	B	.	.	.	0.23	0.11	.	.	.	0.30	0.58
Leu	114	.	.	B	B	.	.	.	0.16	0.06	.	.	.	0.30	0.58
Arg	115	.	.	B	B	.	.	.	0.22	0.11	.	.	F	0.60	1.20
Arg	116	.	.	B	B	.	.	.	0.58	0.26	.	.	F	0.60	1.63
Thr	117	.	.	B	B	.	.	.	0.31	0.33	.	.	F	0.60	1.06
Pro	118	.	.	B	B	.	.	.	0.20	0.51	*	.	F	1.00	0.55
Ala	119	.	.	B	.	.	.	.	0.67	0.09	.	*	.	1.00	0.28
Cys	120	.	.	B	.	.	T	.	0.67	0.34	.	*	.	0.85	0.19
Ala	121	.	.	.	.	T	T	.	0.26	0.14	*	*	.	2.10	0.24
Gly	122	.	.	.	.	T	T	.	0.29	0.19	*	.	F	2.50	0.32
Gly	123	.	.	.	.	T	T	.	0.32	0.04	*	.	F	2.25	0.44
Arg	124	.	.	B	B	.	.	.	0.04	0.14	.	.	F	0.60	0.69
Ser	125	.	.	B	B	.	.	.	0.62	0.13	.	.	F	0.35	1.00
Val	126	.	.	B	B	.	.	.	0.62	0.30	.	.	.	0.70	1.75
Tyr	127	.	.	B	.	.	.	.	0.72	0.23	.	.	.	0.50	0.90
Thr	128	.	.	B	.	.	.	.	0.21	0.53	.	.	.	0.25	1.05
Glu	129	.	.	B	B	.	.	.	0.21	0.79	.	*	.	0.45	1.05
Ala	130	.	.	B	B	.	.	.	0.80	0.63	.	*	.	0.60	0.97
Tyr	131	.	.	B	B	.	.	.	0.16	0.56	.	*	.	0.60	0.47
Val	132	.	.	B	B	.	.	.	0.77	0.50	.	*	.	0.60	0.42
Thr	133	.	.	B	B	.	.	.	0.80	1.14	.	*	.	0.60	0.31
Ile	134	.	.	B	B	.	.	.	1.47	1.07	.	*	.	0.60	0.20
Pro	135	.	.	B	.	.	T	.	1.19	0.89	.	*	.	0.20	0.14
Val	136	.	.	.	.	T	T	.	1.61	0.73	.	.	.	0.20	0.14
Gly	137	.	.	.	.	T	T	.	1.61	0.81	.	.	.	0.20	0.11
Cys	138	.	.	B	.	.	T	.	1.51	0.77	.	.	.	0.20	0.05
Thr	139	.	.	B	.	.	.	.	0.62	0.77	.	.	.	0.40	0.11
Cys	140	.	.	B	.	.	.	.	0.62	0.13	.	.	.	0.10	0.19
Val	141	.	.	B	.	.	T	.	0.23	0.13	.	.	.	0.10	0.55
Pro	142	.	.	B	.	.	T	.	0.62	0.44	.	.	F	0.85	0.65
Glu	143	.	.	B	.	.	T	.	1.29	0.93	.	.	F	1.30	2.44
Pro	144	A	.	.	.	.	T	.	1.01	1.50	*	.	F	1.30	5.49
Glu	145	A	.	.	.	.	.	.	1.68	1.64	*	.	F	1.10	3.59
Lys	146	A	.	.	.	.	.	.	2.23	2.07	*	.	F	1.10	3.46
Asp	147	A	.	.	.	.	T	.	1.56	1.69	.	.	F	1.30	3.00
Ala	148	A	.	.	.	.	T	.	1.56	1.43	*	.	F	1.30	1.21
Asp	149	A	.	.	.	.	T	.	1.47	1.03	*	.	F	1.15	0.98
Ser	150	A	.	.	.	.	T	.	1.17	0.64	*	.	F	1.15	0.78
Ile	151	A	.	.	.	.	.	.	0.23	0.26	*	*	F	0.80	1.04
Asn	152	.	.	B	.	.	T	.	0.23	0.07	*	.	F	0.85	0.44
Ser	153	.	.	B	.	.	T	.	0.87	0.07	*	.	F	0.85	0.54
Ser	154	.	.	B	.	.	T	.	0.87	0.46	*	*	F	1.00	1.55
Ile	155	A	.	.	.	.	T	.	0.82	0.74	.	*	F	1.30	1.67
Asp	156	A	.	.	.	.	T	.	1.12	0.71	*	*	F	1.30	1.23
Lys	157	A	.	.	.	.	T	.	1.17	0.60	*	.	F	1.15	0.93
Gln	158	A	.	.	.	.	T	.	0.66	0.99	*	.	F	1.30	2.65
Gly	159	.	.	B	.	.	T	.	0.14	0.99	*	.	F	1.30	1.31
Ala	160	.	A	B	.	.	.	.	0.22	.30	*	.	F	0.45	0.54
Lys	161	.	A	B	.	.	.	.	0.12	0.39	.	.	F	0.15	0.26
Leu	162	.	A	B	.	.	.	.	0.38	0.41	.	.	.	0.60	0.26
Leu	163	.	A	B	.	.	.	.	0.38	0.41	.	.	.	0.60	0.39
Leu	164	.	A	B	.	.	.	.	0.03	0.31	.	.	.	0.06	0.32
Gly	165	.	.	B	.	.	T	.	0.03	0.31	.	.	F	0.73	0.64
Pro	166	.	.	.	.	.	T	C	0.29	0.13	.	.	F	1.17	0.78
Asn	167	.	.	.	.	T	T	.	0.07	0.13	.	.	F	2.36	1.47
Asp	168	.	.	.	.	.	T	C	0.40	0.31	.	.	F	2.40	1.50
Ala	169	.	.	.	.	.	.	C	1.00	0.31	.	.	F	1.81	0.96
Pro	170	.	.	.	.	.	T	C	0.96	0.31	.	.	F	1.77	0.92
Ala	171	.	.	.	.	.	T	C	0.78	0.29	.	.	.	1.38	0.71
Gly	172	.	.	.	.	.	T	C	0.39	0.14	.	.	.	0.54	0.90
Pro	173	.	.	B	.	.	T	.	0.00	0.07	.	.	.	0.10	0.74