US 5,589,453 AGrant

Human Rhinovirus Receptor Protein (ICAM-1) That Inhibits Rhinovirus Attachment and Infectivity

Issue Date:1996-12-31

•6 Claims

Abstract

A water soluble human rhinovirus (HRV) major receptor preparation comprising detergent-complexed glycoprotein isolated from animal cells, preferably mammalian cells, that express the HRV major receptor and which exhibits the ability to bind to HRV capsids to substantially reduce infectivity of the virus. The purified, water soluble receptor is obtained by extracting cells expressing the receptor with detergent and isolating the solubilized detergent-glycoprotein complexes by binding to monoclonal antibody selective for the HRV receptor protein. Human rhinovirus receptor protein has subsequently been discovered by Greve et al. to be ICAM-1.

Metadata

Assignee

Molecular Therapeutics, Inc.

Inventor

Jeffrey M. Greve

Application Information

Application Number:US 3163830

Filing Date:1994-09-30

Priority Date:1988-09-01

Art Unit:186

Classifications

IPC:

A61K 3817A61K 3900C07K 14705

Field of Search:

424514530435184.1;185.1;198.12;8;21350;395;827;868;380;82769.1;69.3

Patent Drawings

This patent does not have any drawings.

Description

BACKGROUND OF THE INVENTION

The present invention relates to the isolation of proteins from animal cells, particularly mammalian cells, that bind to human rhinovirus (HRV). More particularly, the invention relates to the isolation of HRV receptor proteins that can bind to HRV and thereby block the infectivity of the virus. This property can serve as a basis for inhibiting the initiation or the spread of HRV infections, better known as the common cold.

In order to infect host cells, viruses must bind to and then enter cells to initiate an infection. Since 1959, evidence has accumulated in the literature indicating that the presence of specific binding sites (receptors) on host cells could be a major determinant of tissue tropism of certain viruses. [Holland, J. J., and McLaren, L. C., The mammalian cell-virus relationship, II. Absorption, reception, and eclipse of poliovirus by HeLa cells, J. Exp. Med. 109, 487-504 (1959). Holland, J. J., Receptor affinities as major determinants of enterovirus tissue tropisms in humans, Virology 15, 312-326 (1961).] Among picornaviruses such as poliovirus, coxsacchie virus, and rhinoviruses, specific binding to host cells has been demonstrated. By competition experiments, it has been demonstrated that some of these receptors are distinct from one another in that the saturation of the receptor of one virus had no effect on the binding of a second virus. [Lonberg-Holm, K, Crowell, R. L., and Philipson, L. Unrelated animal viruses share receptors, Nature 259, 679-681 (1976)].

Rhinoviruses form the largest family of picornaviruses, with 115 distinct serotypes identified to date. A large fraction of rhinoviruses (estimated to be 80%) appear to bind to a common receptor on human cells. [Abraham, G., and Colonno, R. J., Many rhinovirus serotypes share the same cellular receptor, J. of Virology 51, 340-345 (1984).] In 1985, the isolation of a monoclonal antibody that appeared to be directed against the major rhinovirus receptor was described. [Colonno, R. J., Callahan, P. L., and Long, W. J., Isolation of a monoclonal antibody that blocks attachment of the major group of human rhinoviruses, J. of Virology 57, 7-12 (1986).] It inhibited infection of cells with the appropriate serotypes of rhinovirus and it inhibited binding of radiolabeled rhinovirus to cells. This group subsequently reported that the monoclonal antibody bound to a protein with an apparent molecular weight of 90,000 daltons. Tomassini, J. E., and Colonno, R. J., Isolation of a receptor protein involved in attachment of human rhinoviruses, J. of Virology 58, 290-295 (1986). This monoclonal antibody has been utilized in clinical trials with primates and humans and is understood to provide some protection against rhinovirus infection.

There are several other reports of attempts at therapeutic intervention in rhinovirus infections. Intranasal application of interferon in humans has been attempted. [Douglas, R. M., et al., Prophylactic efficacy of intranasal alpha2-interferon against rhinovirus infections in the family setting, The New England J. of Medicine, 314, 65-75 (1986).] In this case, significant reduction in the severity of the infection was found, although nosebleeds were observed as a side-effect. Also, several analogs of disoxaril ("WIN" compounds) that reduce the infectivity of a number of picornaviruses (with widely varying effectiveness, depending on the serotype) have been tested in tissue culture and in some animal models. [Fox, M. P., Otto, M. J., and McKinlay, M. A., Antimicrob. Ag. and Chemotherapy 30, 110-116 (1986).] These compounds appear to inhibit replication at a step subsequent to receptor binding, probably at some step of virus uncoating. The atomic coordinates of the binding sites of these compounds within the viral capsid of the serotype HRV14 have been determined by x-ray crystallography, and are located in a hydrophobic pocket present in each protomeric unit of the capsid. [Smith, T. J., et al., The site of attachment in human rhinovirus 14 for antiviral agents that inhibit uncoating, Science 233, 1286-1293 (1986).] The specific function of the binding pocket, if any, is unknown, but drug-resistant mutants with single amino acid interchanges in this region arise at high frequency and are viable. [Badger, J. et al., Structural analysis of a series of antiviral agents complexed with human rhinovirus 14, PNAS 85, 3304-3308 (1988).] This result calls into question the efficacy of such compounds as drugs. The production of anti-peptide antibodies in rabbits has been reported using peptides derived from amino acid sequence of the viral capsid proteins that line the "receptor canyon" of HRV14. [McCray, J., and Werner, G., Different rhinovirus serotypes neutralized by antipeptide antibodies, Nature 329:736-738 (1987).] While the titers of these sera are quite low, cross-serotype protection of cells in tissue culture from rhinovirus infection was demonstrated, raising the possibility of a vaccine.

It is an object of the present invention to isolate an HRV receptor protein from cells having the property of blocking HRV infection. Given the high affinity the virus has for its receptor, it was hypothesized that a therapeutic agent effective against HRV infection might be the receptor itself, or more specifically, the virus binding domain of the receptor. A protein, protein fragment, or peptide that comprises the virus binding domain could block the ability of virus to bind to host cells by occupying (blocking) the receptor binding cleft on the virus. Furthermore, since such a molecule would make some or all of the molecular contacts with the virus capsid that the receptor does, virus mutations that adversely affect binding of the molecule would adversely affect binding of the receptor, and would thus be deleterious or lethal for the virus; therefore, the likelihood of drug-resistant mutants would be very low. Furthermore, such a molecule would be human, lowering the likelihood of being antigenic in humans.

SUMMARY OF THE INVENTION

It has been found that the human rhinovirus (HRV) major receptor can be isolated as a water soluble preparation which exhibits the desired property of binding to HRV capsids and substantially reducing infectivity of the virus. The preparation is in the form of detergent-complexed glycoprotein isolated from animal cells, preferably mammalian cells, that express the HRV major receptor. The purified receptor protein is characterized as follows. It is a glycoprotein with an apparent molecular weight of 95,000 daltons and includes the binding site for HRV. The glycoprotein contains 6-7 asparagine-linked oligosaccharide chains and exists in the preparation in the form of a detergent micelie-bound protein.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

In general terms, the HRV major receptor preparation of the present invention can be obtained by extraction of appropriate animal cells that are known to express the HRV major receptor with a nonionic detergent, followed by immunopurification. Many human cell lines express the receptor, such as HeLa and WI38. Any of these human sources of HRV receptor can be extracted. Particularly useful are HeLa cells. Furthermore, non-human mammalian transfectant cell lines that express the HRV receptor are known or can be prepared which provide another useful source of the receptor. In particular, transfectant cell lines as described in copending U.S. patent application Ser. No. 130,378, infra, provide a ready source of receptor, particularly those secondary transfectants that have been selected for overexpression of receptor. Other animal cells as are known in the art or developed hereafter, such as insect tissue culture cells that have been tranfected with the gene and express the receptor, can also be used.

Essentially any nonionic detergent can be used for the extraction provided the native conformation of the protein receptor is not destroyed. Denaturation of the receptor can be determined by monitoring the ability of the extracted protein to inhibit virus infectivity or by sensitivity to proteolysis. It has been determined that the receptor can be denatured by heating at 60.degree. C. for 30 minutes or by treatment with 1% SDS indicating that care need be taken to maintain the native conformation of the HRV binding site. Examples of useful non-ionic detergents are the alkyl polyoxyethylene ethers (such as Brij), alkylphenyl polyoxyethelene ethers (such as Triton X-100 and Nonidet P-40), acyl polyoxyethylene sorbitan esters (such as Tween), and beta-D-alkyl glucosides, with Triton X-100 being considered particularly preferred.

The key step in the purification of the receptor is fractionation with highly selective anti-receptor antibody. The most ready means to obtain such an antibody is by monoclonal techniques. It is particularly preferred to produce mouse monoclonal antibodies by generating hybridoma cell lines from fusion of murine myeloma cells and mouse transfectant cells expressing the HRV receptor. Further details are available in copending U.S. patent application Ser. No. 130,378, infra. After binding the detergent-glycoprotein complexes obtained from the cell extract to the selected monoclonal antibody, complexes bound to antibody are separated from the remainder of the mixture. Thereafter, detergent-receptor complexes bound to antibody are dissociated, taking steps to again prevent denaturation, and the resulting water soluble receptor preparation isolated. Appropriate conditions for dissociating detergent-receptor complexes from the antibody can be determined empirically and can be expected to vary somewhat from antibody to antibody. Dissociation by raising pH has been found in some cases to be most effective with low pH or high salt conditions being operable but producing lower protein yields.

It is preferable to perform an intermediary purification before purification with antibody. Such intermediary steps comprise adsorbing the detergent extracted protein complexes to a lectin capable of binding HRV receptor, separating absorbed complexes from the remainder of the mixture, and dissociating such complexes for subsequent treatment with antibody. The selection of lectin and dissociating conditions is usually empirical. It has been found that the HRV receptor binds suitably to wheat germ agglutinin lectin and is dissociated effectively by washing with a solution of N-acetyl glucosamine. Because the oligosaccharides on the receptor protein are not completely characterized, and because the receptor protein can be glycosylated differently on different cell types (e.g., mouse cell transfectants), other lectins would be expected also to be suitable. The selection of an appropriate alternative to wheat germ agglutinin and/or eluting agent can be left to the ordinary skill in the art.

The resulting preparation can be treated with proteolytic agents such as proteases, e.g., trypsin, to produce smaller glycoprotein fragments that retain the ability to bind and reduce infectivity of HRV. For example, peptide fragments can be cleaved from a terminal region of the glycoprotein, e.g., the C-terminus, to yield glycoprotein fragments that retain HRV binding. Such glycoprotein fragments can, for example, have apparent molecular weights of between about 80,000 daltons and about 95,000 daltons. Smaller fragments which retain the HRV binding domain of the receptor are also considered to be within the scope of the present invention.

The receptor preparation of the present invention has been shown to inhibit the infectivity of the virus, presumably by binding to the HRV capsid to block its ability then to bind and infect human cells. Such an observation indicates that the receptor preparation will be useful in reducing the infection of host human cells in vivo by contacting the virus with the preparation under conditions favorable to binding with the virus. A therapeutic form would be that of an aqueous solution of the receptor in the presence of nonionic detergent to maintain the receptor in solution and in its native conformation. Detergents with lower critical micelle concentrations, such as the alkyl polyoxyethylene ether Brij 58, would be preferred in order to reduce the concentration of the detergent in the therapeutic solution. The receptor preparation can be administered in vivo by appropriate contact with those areas of the body susceptible to infection by HRV, e.g., by intranasal spray.

The present invention will now be illustrated, but is not intended to be limited, by the following examples.

Preparation of Purified Human Rhinovirus Receptor (HRR) Protein

(1) Human cells (for example, Hela) or mouse L-cell transfectants (for example, the cell lines described in U.S. patent application Ser. No. 130,378, filed Dec. 8, 1987, McClelland and Meyer, "Transfectant Cell Lines Which Express the Major Human Rhinovirus Receptor") were grown up in large numbers as cellular monolayers in standard tissue culture medium (Dulbecco's modified essential medium containing 10% fetal bovine serum; transfectant cells were maintained in the same medium containing HAT (hypoxanthanine/aminoptherin/thymidine) to maintain selective pressure for the selectable marker (Herpes TK). Cells were solubilized for 1 hour at 4.degree. C. in a physiological buffer (Phosphate-buffered saline) containing a nonionic detergent (for example, Triton X-100) (T buffer) and a cocktail of protease inhibitors (aprotinin, leupeptin at 10 .mu.g/ml, EDTA at 1 mM) to prevent proteolytic degradation of the receptor. Insoluble material was removed by filtration through a 0.22 .mu. filter.

(2) The extract was absorbed onto an affinity resin containing Wheat Germ Agglutinin (WGA) (Sigma Chemical Co., St. Louis, Mo., U.S.A) crosslinked to Sepharose for 18 hours at 4.degree. C. with gentle mixing (2 ml packed resin, containing 5 mg WGA/ml resin, per 10.sup.9 cells). The affinity resin was then washed extensively with buffer to remove unbound glycoproteins and eluted with the competing monosaccharide N-acetyl glucosamine (0.3M N-acetyl glucosamine in T buffer) for 1 hour at room temperature.

(3) The WGA-Sepharose eluant is then absorbed to an affinity resin to which purified monoclonal antibody to the HRR has been coupled (e.g., ATCC HB 9a594, referred to in the McClelland and Meyer patent application, Ser. No. 130,378, supra). The monoclonal antibody IgG was purified by ammonium sulfate precipitation [Parham, P., Meth. Enzymol. 92:110-138 (1983)], followed by affinity chromatography on either protein A Sepharose [Ey, P. L., et al., Immunochem. 15:429-436 (1978)] or an Abx column [J. T. Baker Co., Phillipsburg, N.J., U.S.A] following the procedure described by the manufacturer. Monoclonal IgG affinity resin is prepared by coupling IgG to cyanogen bromide-activated Sepharose [Parham, P., supra].

After adding 10 .mu.g/ml human transferrin to block adsorption of transferrin receptor to the resin, the eluant is incubated at 4.degree. C. for 18 hours with the resin with mixing (40-200 .mu.l of resin, containing 5 mg IgG/ml resin, per 10.sup.9 cells), washed extensively with T buffer to remove unbound proteins, and then eluted under nondenaturing conditions with a high pH buffer (0.05M diethanolamine (pH 11.5) with 0.1% Triton X-100) for 1 hour at room temperature. The eluant is removed, neutralized by the addition of 0.2 volumes of 1M HEPES (pH 7.2), and dialysed against three changes of a physiological buffer containing a small amount of nonionic detergent to maintain the solubility of the receptor (0.01M HEPES, 0.150M NaCl, 0.001M CaCl.sub.2, 0.1% Triton X-100, pH 7.5).

The receptor may be further purified by velocity sedimentation through sucrose gradients to remove a group of minor high molecular weight (>200,000 daltons) contaminants. The receptor preparation is layered on top of a 15-35% sucrose gradient (total volume about 4.5 ml, and centrifuged at 300,000.times.g for 18 hours at 4.degree. C. Fractions are collected from the gradient and fractions containing the rhinovirus receptor, which sediments about 1/3 of the way down the gradient, are pooled, concentrated (if necessary), and dialysed.

(4) The resultant preparation from Hela cells was found to contain a glycoprotein with an apparent molecular weight of 95,000 daltons. From mouse transfectant cells, a protein of the same molecular weight but of greater heterogenity (upon analysis by SDS-PAGE) was isolated. The isolated protein has been shown to comprise the rhinovirus receptor by:

(a) Immunoprecipitation from .sup.125 I-surface labeled Hela cells and mouse transfectants expressing the human rhinovirus receptor with a monoclonal antibody that inhibits rhinovirus binding to cells.

(b) Immunoprecitation of purified, .sup.125 I-labeled receptor with the ATCC HB 9594 monoclonal antibody.

(5) A tryptic fragment was prepared by digesting the receptor with 1% (wt E/wt receptor protein) trypsin for 1 hour at 37.degree. C. The reaction mixture was applied to a GF-450 gel filtration column (Dupont) equilibrated in N buffer and the proteolytic fragment separated from the enzyme. Analysis of the resultant fragments by SDS-PAGE indicated a mixture of a 90,000 dalton and an 83,000 dalton fragment of the receptor. These fragments eluted in the same position on a gel filtration column as intact receptor, suggesting that it is bound to a detergent micelle. Amino acid sequencing of the fragments yielded no sequence, indicating that they, like the intact receptor, have a blocked N-terminus, and further indicating that peptides lost from the 90,000 and 83,000 dalton fragments are from the C-terminus of the protein.

Characterization of the Preparation

(1) The purity of the receptor preparation was assessed by SDS-PAGE followed by silver staining. Quantitation of protein was determined by comparing silver stained protein with a series of standard proteins of known amount on SDS-PAGE and confirmed by amino acid analysis, assuming a protein molecular weight of 50,000 daltons (determined by determining the apparent molecular weight on SDS-PAGE of deglycosylated receptor).

(2) The protein was shown to be a glycoprotein containing 6-7 asparagine-linked oligosaccharide chains by digestion of core-glycosylated receptor with endoglycosidase H. Upon gel filtration, the receptor eluted with a volume consistent with a protein molecular weight of 250,000 daltons. This data, along with evidence from chemical cross-linking experiments indicating the receptor is a monomer, are consistent with the receptor behaving like a protein bound to a detergent micelle.

(3) The purified receptor protein was shown to bind to rhinovirus in vitro. When incubated for 30 minutes at 34.degree. C. with 1 .mu.g/ml HRV14 or HRV3, unlabeled, .sup.125 I-labeled, and .sup.35 S-cysteine metabolically labeled HRR could be shown to associate with virus by sedimentation in sucrose gradients or by pelleting in a high speed centrifuge. This binding could be shown to be specific by competing the binding of radiolabeled receptor with unlabeled receptor. The in vitro reaction had the same temperature-dependency as in vivo: receptor bound to the virus at 37.degree. C. but not at 4.degree. C.

(4) The receptor was shown to inhibit infectivity of rhinovirus by incubating HRR with virus (under the same conditions as described above in which binding could be demonstrated) and then testing the resultant mixtures for infectivity by a standard limiting dilution infectivity assay. A Hela cell suspension was prepared by detaching with 0.03% EDTA/PBS for 10 minutes, and the cells washed in 2% FBS/DMEM (I medium) with 10 mM HEPES and adjusted to a concentration of 1.1.times.10.sup.7 cells/ml. Virus or virus-receptor mixtures were serially diluted in I medium, and 20 .mu.l of virus was mixed with 180 .mu.l of cells and incubated for 60 minutes at room temperature. The mixture was then diluted with 9 volumes of I medium and plated out into 8-10 wells of a 96 well tissue culture plate (approximately 200 .mu.l/well), and cultured at 34.degree. C. for 5 days. Cultures were then scored by CPE (cytopathic effect) and the titer of the original stock determined by the following formula:

The results are shown in the Table below.

Additional HRV serotypes were tested. HRV 4, 11, 17 and 89 serotypes (major class) were inhibited by the virus, whereas HRV 1a and 2 (minor class) were not.

The results described above indicate that the purified HRR can block the infectivity of rhinoviruses belonging to the major receptor class of rhinoviruses. The infectivity inhibition property of the receptor protein is correlated with its ability to bind to the virus, and is presumed to act by blocking the receptor binding site on the virus. This property of the receptor is manifested at low concentrations of the receptor protein, and indicates a high affinity of the receptor for the virus. The significance of these results is that the purified, soluble receptor could be used to inhibit the initiation or the spread of rhinovirus infections in vivo. The purified protein also provides a source of material from which smaller protein fragments and peptides could be derived which have the same activity as the intact receptor.

Claims

What is claimed is:

1. A method for reducing the infection by human rhinovirus (HRV) of a host cell susceptible to infection by HRV, comprising contacting the virus under conditions favorable for binding with an antiviral agent selected from the group consisting of human rhinovirus major receptor protein (HRR) and fragments thereof in a form that exhibits the ability to bind to HRV capsids and reduce infectivity of the virus.

2. The method of claim 1 wherein said method is performed in vivo.

3. The method of claim 1 wherein said antiviral agent is isolated from cells that express the human rhinovirus major receptor.

4. The method of claim 3 wherein said antiviral agent is obtained by detergent extraction of said cells.

5. The method of claim 4 wherein said antiviral agent is complexed with a detergent.

6. The method of claim 1 wherein said antiviral agent is purified by immunopurification prior to contact with the virus.

Patent Citations (48)

Patent	Date	Inventor
US4171365	1979-10-01	Diana et al.
US4209526	1980-06-01	Diana et al.
US4232161	1980-11-01	Diana et al.
US4234725	1980-11-01	Diana et al.
US4261928	1981-04-01	Diana et al.
US4372976	1983-02-01	Diana
US4427653	1984-01-01	Springer
US4451476	1984-05-01	Diana
US4843087	1989-06-01	Diana
US4956281	1990-09-01	Wallner et al.
US5109123	1992-04-01	Reinherz et al.
US5235049	1993-08-01	McClelland et al.
AUX14630/88	1988-10-01
AUX48767/90	1990-02-01
EPX169146A3	1986-01-01
EPX169729A2	1986-01-01
EPX192175A2	1986-08-01
EPX207453A2	1987-01-01
EPX227604A2	1987-07-01
EPX261403A2	1988-03-01
EPX280578A2	1988-08-01
EPX287076B1	1988-10-01
EPX289949A2	1988-11-01
EPX314863A2	1989-05-01
EPX319815A2	1989-06-01
EPX380068A1	1990-01-01
EPX364690A2	1990-04-01
EPX362531A1	1990-04-01
EPX362526A2	1990-04-01
EPX365837A2	1990-05-01
EPX379904A1	1990-08-01
EPX387701B1	1990-09-01
EPX387668A1	1990-09-01
EPX391088A2	1990-10-01
EPX459577A2	1991-12-01
DEX3712678A1	1988-10-01
SAX900469	1990-10-01
GBX2022826	1979-12-01
WOXWO88/06592	1988-09-01
WOXWO89/10938	1989-11-01
WOXWO90/03400	1990-04-01
WOXWO90/10646	1990-09-01
WOXWO90/10652	1990-09-01
WOXWO90/13316	1990-11-01
WOXWO91/18010	1991-11-01
WOXWO91/18011	1991-11-01
WOXWO91/16928	1991-11-01
WOXWO91/16927	1991-11-01

Non-Patent Literature (166)

Plow, E. F. et al., Proc. Nat'l Acad. Sci. (USA) 82:8057-8061, "The effect of Arg-Gly-Asp-containing peptides on fibrinogen and von Willebrand factor binding to platelets".
Braude, A. (ed.s), "Infectious Diseases and Medical Microbiology", 2nd edition, W. B. Saunders Co., Philadelphia, PA, (1986) chapter 65 Picornaviruses, pp. 521-529.
Gennaro, A. R. (ed.), Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Co., Easton, PA (1990), "Drug Absorption, Action and Disposition", pp. 707-721.
Martin et al., "Efficient Neutralization and Disruption of Rhinovirus by Chimeric ICAM-1/Immunoglobulin Molecules", J. Virology, 67(6):3561-3568 (Jun. 1993).
Hendley et al., "Transmission of Rhinovirus Colds By Self-Inoculation", The New England Journal of Medicine, 288(26):1361-1364 (Jun. 28, 1973).
Hendley, J. O., and Gwaltney, J. M., Jr., "Mechanisms of Transmission of Rhinovirus Infections", Epidemiologic Reviews, 10:242-257 (1988).
Suter, David, Associated Press, "Tests for a Nasal Spray to Deflect Cold Viruses", New York Times, Sep. 20, 1995.
Manning, Anita, "War on Bacteria Mix of Victories Amid Warnings", USA Today, Sep. 20, 1995.
Haney, Daniel Q., "Beyond Chicken Soup. Nasal Spray Keeps Chimps From Catching Cold Virus", St. Louis Post Dispatch, Sep. 20, 1995.
Associated Press, "Common Colds: Nasal Spray May Help Keep The Sniffles Away", Atlanta Constitution, Sep. 20, 1995.
Associated Press, "Drug Sprays Away Colds", New York Post, Sep. 20, 1995.
Haney, Daniel Q., Associated Press, "The Cold War: Scientists Develop Spray That May End Sniffles", Arizona Republic, Sep. 20, 1995.
Haney, Daniel Q., Associated Press, "For Colds, Nasal Spray Holds Hope. A Protein Swamps The Virus With Potential Targets In The Nose. Its a Decoy Trick", Philadelphia Inquirer, Sep. 20, 1995.
Associated Press, "Simple Nasal Spray May Be Able To Keep Common Cold Away. Medicine Successful On Chimps So Far", Washington Times, Sep. 20, 1995.
Associated Press, "Doctors Sniffing Out Spray to Fight Colds", Denver Post, Sep. 20, 1995.
Associated Press, "Someday Soon, A Simple Sniff Should Snuff The Sniffles", Houston Chronicle, Sep. 20, 1995.
Haney, Daniel Q., Associated Press, "Spray May Ward Off Sniffles. Nasal Treatment Studied To Keep Cold Viruses From Invading Victim", Denver-Rocky Mountain News, Sep. 20, 1995.
Haney, Daniel Q., Associated Press, "Scientists Make Headway In Cold War With Nose Spray", Chicago Sun-Times, Sep. 20, 1995.
Haney, Daniel Q., Associated Press, "Labs Busy Working On Nose Spray To Keep Colds Away", Charlotte Observer, Sep. 20, 1995.
Associated Press, "Nasal Spray May Prevent Sniffles", Miami Herald, Sep. 20, 1995.
Associated Press, "Cure For The Cold? No, But Prevention May Be Spray Away", San Diego Union-Tribune, Sep. 20, 1995.
Haney, Daniel Q., "Nasal Spray Touted As Next-Best Thing To Cure For Colds", The Montreal Gazette, Sep. 20, 1995.
Associated Press, "Scientists Feel They Can Develop Spray To Keep The Sniffles Away", The Spectator, Sep. 20, 1995.
Haney, Daniel Q., Associated Press, "New Nasal Spray May Take Sniffles Out Of Common Cold", Cleveland Plain Dealer, Sep. 20, 1995.
Associated Press, "No Cure, But Nothing To Sniff(le) At. Nasal Spray To Block Common Cold Is In The Works", Minneapolis Star Tribune, Sep. 20, 1995.
Haney, Daniel Q., Associated Press, "Out Front: Progress On Cold Front. Spray May Ward Off Sniffles. Medicine Is First To Block Infection", Sep. 19, 1995.
Monitoring Report, "Cure For Colds Sep. 18 to Sep. 20", Video Monitoring Services of America, a Burrelle's Affiliate, New York, New York, pp. 1-3, Sep. 20, 1995.
Steis, R. G., L. Marcon, J. Clark, W. Urba, D. L. Longo, D. L. Nelson, and A. E. Maluish, "Serum soluble IL-2 receptor as a tumor marker in patients with hairy cell leukemia", Blood (May 1988) 7195:1304-9.
Harning, R., E. Mainofli, J. C. Bystryn, M. Henn, V. J. Merluzzi, and R. Rothlein, "Serum levels of circulating intercellular adhesion molecule 1 in human malignant melanoma", Cancer Res. (1991) 51(8):5003-5.
Shipkowitz, N. L., R. R. Bower, J. B. Schleicher, F. Aquino, R. N. Appell, and W. R. Broderick, "Antiviral Activity of a bis-Benzimidazole Against Experimental Rhinovirus Infections in Chimpanzees", App. Microbiol. (1972) 23(1):117-122.
Sundquist, B., K. L ovgren, S. H oglund, and B. Morein, "Influenza virus ISCOMs: biochemical characterization", Vaccine (Feb. 1988) 6:44-48.
Couch et al., "Effect of Route Inoculation on Experimental Respiratory Viral Disease in Volunteers and Evidence for Airborne Transmission", Bacteriol. Rev. 30:517-529 (1966).
Crump et al., "In Vitro Inhibitory Activity of Soluble ICAM-1 for the Numbered Serotypes of Human Rhinovirus", Antiviral Chemistry and Chemother. 4(6):323-327 (1993).
D'Alessio et al., "Short-Duration Exposure and the Transmission of Rhinoviral Colds", J. Inf. Dis. 150(2):189-193 (1984).
Dochez et al., "Studies in the Common Cold. IV. Experimental Transmission of the Common Cold to Antropoid Apes and Human Beings by Means of a Filtrable Agent", J. Exp. Med. 52:701-716 (1930).
Douglas, R. G., "Pathogenesis of Rhinovirus Common Colds in Human Volunteers", Annals of Otology, Rhinology and Laryngology 79:563-571 (1970).
Greve et al., "Mechanisms of Receptor-Mediated Rhinovirus Neutralization Defined by Two Soluble Forms of ICAM-1", J. Virology 65:6015-6023 (1991).
Gwaltney et al., Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, N. J. Schmidt and R. W. Evans, Eds, 6th edition. pp. 603, Am Pub. Health. Assoc., Washington D. C. (1989).
Halperin et al., "Exacerbations of Asthma in Adults During Experimental Rhinovirus Infection", Am. Rev. Respir. Dis. 132:976-980 (1985).
Hardy et al., "Intranasal Drug Delivery by Spray and Drops", J. Pharm. Pharmacol. 37:294-297 (1985).
Hayden et al., "Modification of Experimental Rhinovirus Colds by Receptor Blockade" Antiviral Research 9:233-247 (1988).
Helenius, A. and Von Bonsdorff, C. H., "Semliki Forest Virus Membrane Proteins, Preparation and Characterization of Spike Complexes Soluble in Detergent-Free Medium" Biochimica et Biophysica Acta 436:895-899 (1976).
Hendley et al., "Relation Between Naturally acquired Immunity and Infectivity of Two Rhinoviruses in Volunteers", J. Inf. Dis. 125:243-248 (1972).
Illum, L., "The Nasal Delivery of Peptides and Proteins", Trends in Biotech. 9:284-289 (1991).
Johnston et al., "Viruses as Precipitants of Asthma Symptoms. III. Rhinoviurses: Molecular Biology and Prospects for Future Intervention", Clin. Exp. Allergy, 23:237 (1993).
Johnston et al., "Viral Infections in Exacerbations in School Children with Cough or Wheeze: A Longitudinal Study", Am. Rev. Resp. Dis., 145:A546 (1992).
Lemanske et al., "Rhinovirus Upper Respiratory Infection Increases Airway Hyperreactivity and Late Asthmatic Reactions", J. Clin. Invest. 83:1-10 (1989).
Marsh et al., "Interactions of Semliki Forest Virus Spike Glycoprotein Rosettes and Vesicle with Cultures Cells", J. Cell Biology 96:455-461 (1983).
Morein, B., "Potentiation of the Immune Response by Immunization with Antigens in Defined Multimeric Physical Forms", Veterinary Immunology and Immunopathology 17:153-159 (1987).
Ohlin et al., "Spectrum of Activity of Soluble Intercellular Adhesion Molecule-1 Against Rhinovirus Reference Strains and Field Isolates", Antimicrob. Agents and Chemother. 38:1413-1415 (1994).
Schipper et al., "The Nasal Mucocilliary Clearance: Relevance to Nasal Drug Delivery", Pharm. Res. 8:807-814 (1991).
Simons et al., "Formation of Protein Micelles from Amphiphilic Membrane Proteins", PNAS 75(11):5306-5310 (1978).
Sundquist et al., "Influenza Virus ISCOMs: Antibody Response in Animals", Vaccine 6:49-53 (1988).
Traunecker et al., "Soluble CD4 Molecules Neutralize Human Immunodeficiency Virus Type 1" Nature 331:84-86 (1988).
Turner at el., "Efficacy of Oral WIN 54954 for Prophylaxis of Experimental Rhinovirus Infection", 37:297-300 (1993).
Woods et al., "In Vitro and In Vivo Activities of WIN 54954, a New Broad Spectrum Antipicornavirus Drug", Antimicrob. Agents Chemother 33:2069-2074 (1989).
Abraham, G. and Colonno, R. J., "Many Rhinovirus Serotypes Share the Same Cellular Receptor", J. Virol. 51:340-345 (1984).
Anasetti et al., "Activation of Natural Killer Cells by LFA-3 Binding to CD2", Publication, Fred Hutchinson Cancer Research Center, Seattle WA, and Molecular Diagnostics, West Haven, CT (U.S.A.).
Argenbright et al., "Monoclonal Antibodies to the Leukocyte Membrane CD18 Glycoprotein Complex and to Intercellular Adhesion Molecule-1 Inhibit Leukocyte-Endothelial Adhesion in Rabbits", J. Leukoc. Biol. 49:253-257 (1991).
Argenbright, L. W. and Barton, R. W., "Interactions of Leukocyte Integrins with Intercellular Adhesion Molecule-1 in the Production of Inflammatory Vascular Injury In Vivo: the Shwartzmann Reaction Revisited", J. Clin. Invest. 89(1):259-272 (1992).
Badger et. al., "Structure Analysis of a Series of Antiviral Agents Complexed with Human Rhinovirus 14", PNAS 85:3304-3308 (1988).
Bangham, C. R. M. and McMichael, A. J., "Nosing ahead in the cold war" Nature 334:16 (1990).
Bebbington, C. R., and Hentschel, C. C. G. "The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells" DNA Cloning 3:163-186 (1987).
Blann, A. D., "Cell Hybrids: an important new source of antibody production" Med. Lab. Sci. 36:329-338 (1979).
Bock et al., "Characterization of soluble forms of NCAM", FEBS Lett 225 (1,2):33-36 (1987).
Bowie et al., "Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions", Science 247:1306-1310 (1990).
Campbell, B. A. and Cords, C. E., "Monoclonal Antibodies That Inhibit Attachment of Group B Coxsackieviruses", J. Virol. 48(2):561-564 (1983).
Capon et al., "Designing CD4 immunoadhesins for AIDS therapy", Nature 337:525-531 (1989).
Cate et al., "Isolation of the Bovine and Human Genes for M ullerian Inhibiting Substance and Expression of the Human Gene in Animal Cells", Cell 45:685-698 (1986).
Cole et al., "Topographic Localization of the Heparin-binding Domain of the Neural Cell Adhesion Molecule N-CAM", J. Cell Biol. 103:1739-1744 (1986).
Colonno, et al., "Isolation of a Monoclonal Antibody that Blocks Attachment of the Major Group of Human Rhinoviruses", J. Virology 57: 7-12 (1986).
Colonno, R. J. and Tomassini, J. E., "Viral Receptors: A Novel Approach For The Prevention Of Human Rhinovirus Infection", in Medical Virology VI, de la Maza, L. M. and E. M. Peterson, eds. (Elsevier, New York, 1987) 331-351.
Cooper, G. M., "Cellular Transforming Genes", Science 217: 801-806 (1982).
Couch, R. B., "Rhinoviruses", Virology, Second Edition, edited by B. N. Fields, D. M. Knipe et al. Raven Press, Ltd., New York, 607-629 (1990).
Creighton, T. E., Proteins by W. H. Freeman and Company, New York, 33-34 (1984).
Cunningham et al., "Neural Cell Adhesion Molecule: Structure, Immunoglobulin-Like Domains, Cell Surface Modulation, and Alternative RNA Splicing", Science 236:799-806 (1987).
Cybulsky, M. I. and Gimbrone, Jr., M. A., "Endothelial Expression of a Mononuclear Leukocyte Adhesion Molecule During Atherogenesis", Science 251:788-791 (1991).
Deen et al., "A Soluble Form of CD4 (T4) Protein Inhibits AIDS Virus Infection", Nature 331:82-86 (1988).
Dick, E. C., "Experimental Infection of Chimpanzees with Human Rhinovirus Types 14 and 43", Proceedings Of The Society For Experimental Biology And Medicine 127:1079-1081 (1968).
Douglas et al., "Prophylactic Efficacy of Intranasal Alpha 2-Interferon Against Rhinovirus Infections in the Family Setting", The New England J. of Med. 314:65-70 (1986).
Dustin et al., "Induction by IL 1 and Interferon-.gamma.: Tissue Distribution, Biochemistry, and Function of a Natural Adherence Molecule (ICAM-1)", J. Immunol. 137(1):245-254 (1986).
Dustin et al., "Supergene Families Meet in the Immune System" Immunology Today, 9(7 and 8):213-215 (1988).
Dustin et al., "Correlation of CD2 Binding and Functional Properties of Multimeric and Monomeric Lymphocyte Function-Associated Antigen 3", J. Exp. Med. 169:503-517 (1989).
Ey, P. L., et. al., "Isolation of Pure IgG1, IgG2a, and IgG2b Immunoglobulins from Mouse Serum Using Protein A--Sepharose", Immunochemistry 15:429-436 (1978).
Fisher et al., "HIV Infection is Blocked in vitro by Recombinant Soluble CD4", Nature 331:76-78 (1988).
Fox et al., "Prevention of a Rhinovirus and Poliovirus Uncoating by WIN 51711, a New Antiviral Drug", Antimicrob. Ag. and Chemotherapy 30: 110-116 (1986).
Galfrey et. al., "Antibodies to Major Histocompatibility Antigens Produced by Hybrid Cell Lines", Nature 266:550-552 (1977).
Gething, M. J. and Sambrook, J., "Construction of Influenza Haemagglutinin Genes that Code for Intracellular and Secreted Forms of the Protein" Nature 300:598-603 (1982).
Ginsberg et al., "Inhibition of Fibronectin Binding to Platelets by Proteolytic Fragments and Synthetic Peptides Which Support Fibroblast Adhesion", J. Biol. Chem. 260(7):3931-3936 (1985).
Giranda et al., "Modeling of the Human Intercellular Adhesion Molecule-1, the Human Rhinovirus Major Group Receptor" Proteins: Structure, Function, and Genetics, 7:227-233 (1990).
Gough, N., "Putting A Stop To An Immunoglobulin Message", Trends Genet. 3(9):238-240 (1987).
Gower et al., "Alternative Splicing Generates a Secreted Form of N-CAM in Muscle and Brain", Cell 55:955-964 (1988).
Graham, F. L., and Van der Eb, A. J., "A New Technique for the Assay of Infectivity of Human Adenovirus 5 DNA", Virology 52:456-467 (1973).
Green et al., "Immunogenic Structure of the Influenza Virus Hemagglutinin", Cell 28:477-487 (1982).
Greve et al., "The Major Human Rhinovirus Receptor Is ICAM-1", Cell 56:839-847 (1989).
Gross-Bellard et al., "Isolation of High-Molecular-Weight DNA from Mammalian Cells", Eur. J. Biochem. 36:32-38 (1973).
G ussow, D. and Ploegh, H., "Soluble class I antigens: a conundrum with no solution?", Immunology Today 8(7,8):220-222 (1987).
Hamparian et al., "A Collaborative Report: Rhinoviruses-Extension of the Numbering System from 89 to 100", Virology 159:191-192 (1987).
Hayden et al., "Safety and Efficacy of Intranasal Pirodavir (R77975) in Experimental Rhinovirus Infection", Antimicrob. Agents Chemother. 36(4): 727-732 (1992).
Hayden et al., "Prevention of Natural Colds by Contact Prophylaxis with Intrnasal Alpha2-Interferon", The New England Journal of Medicine, 314(2):71-75 (1986).
Holland, J. J. and McLaren, L. C., "The mammalian cell-virus relationship. II. Absorption, Reception and Eclipse of Poliovirus by HeLa Cells" J. Exp. Med. 109:487-504 (1959).
Holland, J. J., "Receptor affinities as Major Determinants of Enterovirus Tissue Tropisms in Humans", Virology 15:312-326.
Horton et al., "Gene Splicing by Overlap Extension: Tailor-Made Genes Using the Polyerase Chain Reaction", BioTechniques 8(5):528-535 (1990).
Hussey et. al., "A Soluble CD4 Protein Selectively Inhibits HIV Replication and Syncytium Formation" Nature, 331:78-81 (1988).
Kamarck, M. E., and Ruddle, F. H., "Somatic Cell Genetics and the Human Gene Map", Chapter 105 in Handbook of Experimental Immunology in Four Volumes, vol. 3: Genetics and Molecular Immunology, D. M. Weir, ed. (Blackwell Scientific Publications, Boston, MA, 1986).
Katz et al., "Chromosome Mapping of Cell Membrane Antigens Expressed on Activiated B Cells", Eur. J. Immunol., 15:103-106 (1985).
Kavenoff, R., and Zimm, B. H., "Chromosome-Sized DNA Molecules from Drosophila", Chromosoma (Berl.) 41:1-27 (1973).
K uhn et al., "Gene Transfer, Expression, and Molecular Cloning of the Human Transferrin Receptor Gene", Cell 37:95-103 (1984).
Lebman et al., "A Monoclonal Antibody that Detects Expression of Transferrin Receptor in Human Erythroid Precursor Cells", Blood 59(3): 671-678 (1982).
Littlefield, J. W., "Selection of Hybrids from Matings of Fibroblasts in vitro and Their Presumed Recombinants", Science 145:709-710 (1964).
Lonberg-Holm et al., "Unrelated Animal Viruses Share Receptors", Nature 259:679-681 (1976).
Margulies, D. H., et. al., "Engineering Soluble Major Histocompatibility Molecules: Why and How", Immunol. Res. 6: 101-116 (1987).
Marlin, S. D. and Springer, T. A., "Purified Intercellular Adhesion Molecule-1 (ICAM-1) Is a Ligand for Lymphocyte Function-Associated Antigen 1 (LFA-1)", Cell 51:813-819 (1987).
Marlin et al., "A Soluble Form of Intercellular Adhesion Molecule-1 Inhibits Rhinovirus Infection", Nature 344:70-72 (1990).
Marsh et al., "Antibody-toxin Conjugation", Immunotoxins by Kluwer Academic Publishers, Boston, Dordrecht, Lancaster 213-237 (1988).
McClelland et al., "Identification of Monoclonal Antibody Epitopes and Critical Residues for Rhinovirus in Domain 1 of ICAM-1", PNAS 88(18): 7993-7997 (1991).
McClelland et al., "Transfectant cell lines which express the major human rhinovirus receptor, their preparation, and their uses", Immuno. 112: 117179 (1990).
McCray, J. and Werner, G., "Different Rhinovirus Serotypes Neutralized by Antipeptide Antibodies", Nature 329:736-738 (1987).
Medical Microbiology: "An Introduction to Infectious Diseases", 2nd ed., J. C. Sherris, ed. (Elsevier Science Publishing Co., Inc., N.Y. 1990) pp. 514-515.
Medrano, L. and Green, H., "Picornavirus Receptors and Picornavirus Multiplication in Human-Mouse Hybrid Cell Lines", Virology 54:515-524 (1973).
Melchers et al., Lymphocyte Hybridomas, vol. 81 of Current Topics in Microbiology and Immunology, W. Arber, W. Henle, P. H. Hofschneider, J. H. Humphrey, J. Klein, P. Koldovsky, H. Koprowski, O. Maaloe, F. Melchers, R. Rott, H. G. Schweiger, L. Syrucek, P. K. Vogt, eds (Springer Verlang, New York, 1978).
Mendelsohn et al., "Transformation of a Human Poliovirus Receptor Gene into Mouse Cells", PNAS 83:7845-7849 (1986).
Minor, P. D., "Growth, Assay and Purification of Picornaviruses", in Virology: A Practical Approach, B. W. J. Mahy, ed. (IRL Press Limited, Oxford, England), 25-41 (1985).
Minor et al., "Monoclonal antibodies which block cellular receptors of poliovirus", Virus Research 1:203-212 (1984).
Niman et al., "Anti-peptide antibodies detect oncogene-related proteins in urine", PNAS 82:7924-7928 (1985).
Nobis et al., "Production of a Monoclonal Antibody against an Epitope on HeLa Cells that Is the Functional Poliovirus Binding Site", J. Gen. Virol. 66:2563-2569 (1985).
Parham, P., "Monoclonal Antibodies Against HLA Products and Their use in Immunaffinity Purification," Methods in Enzymology 92:110-138 (1983).
Pepinsky et al., "The Increased Potency of Crossed-linked Lymphocyte Function-associated Antigen-3 (LFA-3) Multimers Is a Direct Consequence of Changes in Valency", J. Biol. Chem. 266(27):18244-18249 (1991).
Peterson, A. and Seed, B., "Genetic Analysis of Monoclonal Antibody and HIV Binding Sites on the Human Lymophocyte Antigen CD4", Cell 54:65-72 (1988).
Rossman et al., "Structure of a Human Common Cold Virus and Functional Relationship to other Picornaviruses", Nature 317:145-153 (1985).
Rothlein et al., "A Form of Circulating ICAM-1 In Human Serum", J. Immuno. 147(11):3788-3793 (1991).
Rothlein et al., "A Human Intercellular Adhesion Molecule (ICAM-1) Distinct From LFA-1", J. Immuno. 137(4):1270-1274 (1986).
Ruddle et al., "DNA-Mediated Gene Transfer in Mammalian Gene Cloning", Genetic Engineering 6:319-338 (1984).
Ruoslahti et al., "Synthetic Peptides in the Analysis of Cell Adhesion," in Synthetic Peptides in Biology and Medicine Elsvier Science Publishers, pp. 191-197 (1985).
Saiki et al., "Enzymatic Amplification of .beta.-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia," Science 230:1350-1354 (1985).
Sambrook et al., Molecular Cloning: A Laboratory Manual, 2d ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989) pp. 1.21-1.52.
Scopes, R. K., "Separation By Precipitation," in Protein Purification: Principles & Practice (1982) Springler Verlag, NY, pp. 39-46.
Seed, B. and Aruffo, A., "Molecular Cloning of the CD2 antigen, the T-Cell Erythrocyte Receptor, by a Rapid Immunoselection Procedure," PNAS 84:3365-3369 (1987).
Seed, B., "An LFA-3 cDNA Encodes a Phospholipid-Linked Membrane Protein Homologous to its Receptor CD2," Nature 329:840-842 (1987).
Seth et al., "Circulating ICAM-1 isoforms: Diagnostic Prospects for Inflammatory and Immune Disorders," Lancet 338:83-84 (1991).
Sherman-Gold, R., "Companies Pursue Therapies Based on Complex Cell Adhesion Molecules", Genetic Engineering News pp. 6-7, 14 (Jul. 1993).
Sherry, B. and Rueckert, R., "Evidence for at Least Two Dominant Neutralization Antigens on Human Rhinovirus 14," J. Virol. 53(1): 137-143 (1985).
Shih, C. and Weinberg, R. A., "Isolation of a Transforming Sequence from a Human Bladder Carcinoma Cell Line," Cell 29:161-169 (1982).
Siddique et al., "The Poliovirus Sensitivity (PVS) Gene Is on Chromosome 19q12->q13.2", Genomics 3:156-160 (1988).
Simmons et al., "ICAM, an Adhesion Ligand of LFA-1, is Homologous to the Neural Cell Adhesion Molecule NCAM," Nature 331:624-627 (1988).
Skerra, A. and Pluckthun, A., "Assembly of a Functional Immunoglobulin FV Fragment in Escherichia Coli" Science, 240:1038-1041 (1988).
Smith, T. J., et. al., "The Site of Attachment in Human Rhinovirus 14 for 4 Antiviral Agents that Inhibit Uncoating", Science 233:1286-1293 (1986).
Smith et al., "Blocking of HIV-1 Infectivity by a Soluble, Secreted Form of the CD4 Antigen," Science 238:1704-1707 (1987).
Smith et al., "Modification and Secretion of Human Interleukin 2 Produced in Insect Cells by a Baculovirus Expression Vector", PNAS 82:8404-8408 (1985).
Springer, T. A., "Adhesion Receptors of the Immune System", Nature 346:425-434 (1990).
Staunton et al., "Primary Structure of ICAM-1 Demonstrates Interaction between Members of the Immunoglobulin and Integrin Supergene Families," Cell 52:925-933 (1988).
Staunton et al., "The Arrangement of the Immunoglobulin-Like Domains of ICAM-1 and the Binding Sites for LFA-1 and Rhinovirus," Cell 61:243-254 (1990).
Staunton et al., "A Cell Adhesion Molecule, ICAM-1, is the Major Surface Receptor for Rhinoviruses," Cell 56:849-853 (1989).
Tomassini, J. E., "Isolation, Characterization and Cloning of the Cellular Receptor for the Major Group of Human Rhinoviruses," Ph.D. Thesis, University of Pennsylvania (1986).
Tomassini, J. E. and Colonno, R. J., "Isolation of a Receptor Protein Involved in Attachment of Human Rhinoviruses," J. Virol. 58(2): 290-295 (1986).
Tomassini et al., "cDNA Cloning Reveals that the Major Group Rhinovirus Receptor on HeLa Cells is Intercellular Adhesion Molecule 1," PNAS 86:4907-4911 (1989).
Towbin et. al., "Electrophoretic Transfer of Proteins from Polyacrylamide Gels to Nitrocellulose Sheets: Procedure and Some Applications", PNAS 76(9):4350-4354 (1979).
Traunecker et al., "Highly Efficient Neutralization of HIV with Recombinant CD4-Immunoglobulin Molecules," Nature 339:68-70 (1989).
Urlaub, G. and Chasin, L. A., "Isolation of Chinese Hamster Cell Mutants Deficient in Dihydrofolate Reductase Activity," PNAS USA 77(7): 4216-4220 (1980).
Wade, N., "Hybridomas: A Potent New Biotechnology," Science 208: 692-693 (1980).
Welsh, K. I., "Antibody Production Made Easier," Nature 266: 495 (1977).
Wigler et al., "Transformation of Mammalian Cells with Genes from Procaryotes and Eucaryotes," Cell 16:777-785 (1979).
Williams, A. F., "A Year in the Life of the Immunoglobulin Superfamily", Immunology Today 8(10):298-303 (1987).
Williams, A. F. and Barclay, A. N., "The Immunoglobulin Superfamily-Domains for Cell Surface Recognition.sup.1,2 ", Ann. Rev. Immunol. 6:381-405 (1988).
Winther et al., "Sites of Rhinovirus Recovery After Point Inoculation of the Upper Airway", JAMA 256(13):1763-1767 (1986).
Zettlmeissl et al., "Expression and Characterization of Human CD4: Immunoglobulin Fusion Proteins," DNA and Cell Biology 9:347-353 (1990).

Abstract

Description

BACKGROUND OF THE INVENTION

SUMMARY OF THE INVENTION

DESCRIPTION OF THE PREFERRED EMBODIMENTS

The present invention will now be illustrated, but is not intended to be limited, by the following examples.

Preparation of Purified Human Rhinovirus Receptor (HRR) Protein

(b) Immunoprecitation of purified, .sup.125 I-labeled receptor with the ATCC HB 9594 monoclonal antibody.

Characterization of the Preparation

The results are shown in the Table below.

Additional HRV serotypes were tested. HRV 4, 11, 17 and 89 serotypes (major class) were inhibited by the virus, whereas HRV 1a and 2 (minor class) were not.

Claims

What is claimed is:

2. The method of claim 1 wherein said method is performed in vivo.

3. The method of claim 1 wherein said antiviral agent is isolated from cells that express the human rhinovirus major receptor.

4. The method of claim 3 wherein said antiviral agent is obtained by detergent extraction of said cells.

5. The method of claim 4 wherein said antiviral agent is complexed with a detergent.

6. The method of claim 1 wherein said antiviral agent is purified by immunopurification prior to contact with the virus.

Patent Citations (48)

Patent	Date	Inventor
US4171365	1979-10-01	Diana et al.
US4209526	1980-06-01	Diana et al.
US4232161	1980-11-01	Diana et al.
US4234725	1980-11-01	Diana et al.
US4261928	1981-04-01	Diana et al.
US4372976	1983-02-01	Diana
US4427653	1984-01-01	Springer
US4451476	1984-05-01	Diana
US4843087	1989-06-01	Diana
US4956281	1990-09-01	Wallner et al.
US5109123	1992-04-01	Reinherz et al.
US5235049	1993-08-01	McClelland et al.
AUX14630/88	1988-10-01
AUX48767/90	1990-02-01
EPX169146A3	1986-01-01
EPX169729A2	1986-01-01
EPX192175A2	1986-08-01
EPX207453A2	1987-01-01
EPX227604A2	1987-07-01
EPX261403A2	1988-03-01
EPX280578A2	1988-08-01
EPX287076B1	1988-10-01
EPX289949A2	1988-11-01
EPX314863A2	1989-05-01
EPX319815A2	1989-06-01
EPX380068A1	1990-01-01
EPX364690A2	1990-04-01
EPX362531A1	1990-04-01
EPX362526A2	1990-04-01
EPX365837A2	1990-05-01
EPX379904A1	1990-08-01
EPX387701B1	1990-09-01
EPX387668A1	1990-09-01
EPX391088A2	1990-10-01
EPX459577A2	1991-12-01
DEX3712678A1	1988-10-01
SAX900469	1990-10-01
GBX2022826	1979-12-01
WOXWO88/06592	1988-09-01
WOXWO89/10938	1989-11-01
WOXWO90/03400	1990-04-01
WOXWO90/10646	1990-09-01
WOXWO90/10652	1990-09-01
WOXWO90/13316	1990-11-01
WOXWO91/18010	1991-11-01
WOXWO91/18011	1991-11-01
WOXWO91/16928	1991-11-01
WOXWO91/16927	1991-11-01

Non-Patent Literature (166)

Plow, E. F. et al., Proc. Nat'l Acad. Sci. (USA) 82:8057-8061, "The effect of Arg-Gly-Asp-containing peptides on fibrinogen and von Willebrand factor binding to platelets".

Braude, A. (ed.s), "Infectious Diseases and Medical Microbiology", 2nd edition, W. B. Saunders Co., Philadelphia, PA, (1986) chapter 65 Picornaviruses, pp. 521-529.

Gennaro, A. R. (ed.), Remington's Pharmaceutical Sciences, 18th edition, Mack Publishing Co., Easton, PA (1990), "Drug Absorption, Action and Disposition", pp. 707-721.

Martin et al., "Efficient Neutralization and Disruption of Rhinovirus by Chimeric ICAM-1/Immunoglobulin Molecules", J. Virology, 67(6):3561-3568 (Jun. 1993).

Hendley et al., "Transmission of Rhinovirus Colds By Self-Inoculation", The New England Journal of Medicine, 288(26):1361-1364 (Jun. 28, 1973).

Hendley, J. O., and Gwaltney, J. M., Jr., "Mechanisms of Transmission of Rhinovirus Infections", Epidemiologic Reviews, 10:242-257 (1988).

Suter, David, Associated Press, "Tests for a Nasal Spray to Deflect Cold Viruses", New York Times, Sep. 20, 1995.

Manning, Anita, "War on Bacteria Mix of Victories Amid Warnings", USA Today, Sep. 20, 1995.

Haney, Daniel Q., "Beyond Chicken Soup. Nasal Spray Keeps Chimps From Catching Cold Virus", St. Louis Post Dispatch, Sep. 20, 1995.

Associated Press, "Common Colds: Nasal Spray May Help Keep The Sniffles Away", Atlanta Constitution, Sep. 20, 1995.

Associated Press, "Drug Sprays Away Colds", New York Post, Sep. 20, 1995.

Haney, Daniel Q., Associated Press, "The Cold War: Scientists Develop Spray That May End Sniffles", Arizona Republic, Sep. 20, 1995.

Haney, Daniel Q., Associated Press, "For Colds, Nasal Spray Holds Hope. A Protein Swamps The Virus With Potential Targets In The Nose. Its a Decoy Trick", Philadelphia Inquirer, Sep. 20, 1995.

Associated Press, "Simple Nasal Spray May Be Able To Keep Common Cold Away. Medicine Successful On Chimps So Far", Washington Times, Sep. 20, 1995.

Associated Press, "Doctors Sniffing Out Spray to Fight Colds", Denver Post, Sep. 20, 1995.

Associated Press, "Someday Soon, A Simple Sniff Should Snuff The Sniffles", Houston Chronicle, Sep. 20, 1995.

Haney, Daniel Q., Associated Press, "Spray May Ward Off Sniffles. Nasal Treatment Studied To Keep Cold Viruses From Invading Victim", Denver-Rocky Mountain News, Sep. 20, 1995.

Haney, Daniel Q., Associated Press, "Scientists Make Headway In Cold War With Nose Spray", Chicago Sun-Times, Sep. 20, 1995.

Haney, Daniel Q., Associated Press, "Labs Busy Working On Nose Spray To Keep Colds Away", Charlotte Observer, Sep. 20, 1995.

Associated Press, "Nasal Spray May Prevent Sniffles", Miami Herald, Sep. 20, 1995.

Associated Press, "Cure For The Cold? No, But Prevention May Be Spray Away", San Diego Union-Tribune, Sep. 20, 1995.

Haney, Daniel Q., "Nasal Spray Touted As Next-Best Thing To Cure For Colds", The Montreal Gazette, Sep. 20, 1995.

Associated Press, "Scientists Feel They Can Develop Spray To Keep The Sniffles Away", The Spectator, Sep. 20, 1995.

Haney, Daniel Q., Associated Press, "New Nasal Spray May Take Sniffles Out Of Common Cold", Cleveland Plain Dealer, Sep. 20, 1995.

Associated Press, "No Cure, But Nothing To Sniff(le) At. Nasal Spray To Block Common Cold Is In The Works", Minneapolis Star Tribune, Sep. 20, 1995.

Haney, Daniel Q., Associated Press, "Out Front: Progress On Cold Front. Spray May Ward Off Sniffles. Medicine Is First To Block Infection", Sep. 19, 1995.

Monitoring Report, "Cure For Colds Sep. 18 to Sep. 20", Video Monitoring Services of America, a Burrelle's Affiliate, New York, New York, pp. 1-3, Sep. 20, 1995.

Steis, R. G., L. Marcon, J. Clark, W. Urba, D. L. Longo, D. L. Nelson, and A. E. Maluish, "Serum soluble IL-2 receptor as a tumor marker in patients with hairy cell leukemia", Blood (May 1988) 7195:1304-9.

Harning, R., E. Mainofli, J. C. Bystryn, M. Henn, V. J. Merluzzi, and R. Rothlein, "Serum levels of circulating intercellular adhesion molecule 1 in human malignant melanoma", Cancer Res. (1991) 51(8):5003-5.

Shipkowitz, N. L., R. R. Bower, J. B. Schleicher, F. Aquino, R. N. Appell, and W. R. Broderick, "Antiviral Activity of a bis-Benzimidazole Against Experimental Rhinovirus Infections in Chimpanzees", App. Microbiol. (1972) 23(1):117-122.

Sundquist, B., K. L ovgren, S. H oglund, and B. Morein, "Influenza virus ISCOMs: biochemical characterization", Vaccine (Feb. 1988) 6:44-48.

Couch et al., "Effect of Route Inoculation on Experimental Respiratory Viral Disease in Volunteers and Evidence for Airborne Transmission", Bacteriol. Rev. 30:517-529 (1966).

Crump et al., "In Vitro Inhibitory Activity of Soluble ICAM-1 for the Numbered Serotypes of Human Rhinovirus", Antiviral Chemistry and Chemother. 4(6):323-327 (1993).

D'Alessio et al., "Short-Duration Exposure and the Transmission of Rhinoviral Colds", J. Inf. Dis. 150(2):189-193 (1984).

Dochez et al., "Studies in the Common Cold. IV. Experimental Transmission of the Common Cold to Antropoid Apes and Human Beings by Means of a Filtrable Agent", J. Exp. Med. 52:701-716 (1930).

Douglas, R. G., "Pathogenesis of Rhinovirus Common Colds in Human Volunteers", Annals of Otology, Rhinology and Laryngology 79:563-571 (1970).

Greve et al., "Mechanisms of Receptor-Mediated Rhinovirus Neutralization Defined by Two Soluble Forms of ICAM-1", J. Virology 65:6015-6023 (1991).

Gwaltney et al., Diagnostic Procedures for Viral, Rickettsial and Chlamydial Infections, N. J. Schmidt and R. W. Evans, Eds, 6th edition. pp. 603, Am Pub. Health. Assoc., Washington D. C. (1989).

Halperin et al., "Exacerbations of Asthma in Adults During Experimental Rhinovirus Infection", Am. Rev. Respir. Dis. 132:976-980 (1985).

Hardy et al., "Intranasal Drug Delivery by Spray and Drops", J. Pharm. Pharmacol. 37:294-297 (1985).

Hayden et al., "Modification of Experimental Rhinovirus Colds by Receptor Blockade" Antiviral Research 9:233-247 (1988).

Helenius, A. and Von Bonsdorff, C. H., "Semliki Forest Virus Membrane Proteins, Preparation and Characterization of Spike Complexes Soluble in Detergent-Free Medium" Biochimica et Biophysica Acta 436:895-899 (1976).

Hendley et al., "Relation Between Naturally acquired Immunity and Infectivity of Two Rhinoviruses in Volunteers", J. Inf. Dis. 125:243-248 (1972).

Illum, L., "The Nasal Delivery of Peptides and Proteins", Trends in Biotech. 9:284-289 (1991).

Johnston et al., "Viruses as Precipitants of Asthma Symptoms. III. Rhinoviurses: Molecular Biology and Prospects for Future Intervention", Clin. Exp. Allergy, 23:237 (1993).

Johnston et al., "Viral Infections in Exacerbations in School Children with Cough or Wheeze: A Longitudinal Study", Am. Rev. Resp. Dis., 145:A546 (1992).

Lemanske et al., "Rhinovirus Upper Respiratory Infection Increases Airway Hyperreactivity and Late Asthmatic Reactions", J. Clin. Invest. 83:1-10 (1989).

Marsh et al., "Interactions of Semliki Forest Virus Spike Glycoprotein Rosettes and Vesicle with Cultures Cells", J. Cell Biology 96:455-461 (1983).

Morein, B., "Potentiation of the Immune Response by Immunization with Antigens in Defined Multimeric Physical Forms", Veterinary Immunology and Immunopathology 17:153-159 (1987).

Ohlin et al., "Spectrum of Activity of Soluble Intercellular Adhesion Molecule-1 Against Rhinovirus Reference Strains and Field Isolates", Antimicrob. Agents and Chemother. 38:1413-1415 (1994).

Schipper et al., "The Nasal Mucocilliary Clearance: Relevance to Nasal Drug Delivery", Pharm. Res. 8:807-814 (1991).

Simons et al., "Formation of Protein Micelles from Amphiphilic Membrane Proteins", PNAS 75(11):5306-5310 (1978).

Sundquist et al., "Influenza Virus ISCOMs: Antibody Response in Animals", Vaccine 6:49-53 (1988).

Traunecker et al., "Soluble CD4 Molecules Neutralize Human Immunodeficiency Virus Type 1" Nature 331:84-86 (1988).

Turner at el., "Efficacy of Oral WIN 54954 for Prophylaxis of Experimental Rhinovirus Infection", 37:297-300 (1993).

Woods et al., "In Vitro and In Vivo Activities of WIN 54954, a New Broad Spectrum Antipicornavirus Drug", Antimicrob. Agents Chemother 33:2069-2074 (1989).

Abraham, G. and Colonno, R. J., "Many Rhinovirus Serotypes Share the Same Cellular Receptor", J. Virol. 51:340-345 (1984).

Anasetti et al., "Activation of Natural Killer Cells by LFA-3 Binding to CD2", Publication, Fred Hutchinson Cancer Research Center, Seattle WA, and Molecular Diagnostics, West Haven, CT (U.S.A.).

Argenbright et al., "Monoclonal Antibodies to the Leukocyte Membrane CD18 Glycoprotein Complex and to Intercellular Adhesion Molecule-1 Inhibit Leukocyte-Endothelial Adhesion in Rabbits", J. Leukoc. Biol. 49:253-257 (1991).

Argenbright, L. W. and Barton, R. W., "Interactions of Leukocyte Integrins with Intercellular Adhesion Molecule-1 in the Production of Inflammatory Vascular Injury In Vivo: the Shwartzmann Reaction Revisited", J. Clin. Invest. 89(1):259-272 (1992).

Badger et. al., "Structure Analysis of a Series of Antiviral Agents Complexed with Human Rhinovirus 14", PNAS 85:3304-3308 (1988).

Bangham, C. R. M. and McMichael, A. J., "Nosing ahead in the cold war" Nature 334:16 (1990).

Bebbington, C. R., and Hentschel, C. C. G. "The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells" DNA Cloning 3:163-186 (1987).

Blann, A. D., "Cell Hybrids: an important new source of antibody production" Med. Lab. Sci. 36:329-338 (1979).

Bock et al., "Characterization of soluble forms of NCAM", FEBS Lett 225 (1,2):33-36 (1987).

Bowie et al., "Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions", Science 247:1306-1310 (1990).

Campbell, B. A. and Cords, C. E., "Monoclonal Antibodies That Inhibit Attachment of Group B Coxsackieviruses", J. Virol. 48(2):561-564 (1983).

Capon et al., "Designing CD4 immunoadhesins for AIDS therapy", Nature 337:525-531 (1989).

Cate et al., "Isolation of the Bovine and Human Genes for M ullerian Inhibiting Substance and Expression of the Human Gene in Animal Cells", Cell 45:685-698 (1986).

Cole et al., "Topographic Localization of the Heparin-binding Domain of the Neural Cell Adhesion Molecule N-CAM", J. Cell Biol. 103:1739-1744 (1986).

Colonno, et al., "Isolation of a Monoclonal Antibody that Blocks Attachment of the Major Group of Human Rhinoviruses", J. Virology 57: 7-12 (1986).

Colonno, R. J. and Tomassini, J. E., "Viral Receptors: A Novel Approach For The Prevention Of Human Rhinovirus Infection", in Medical Virology VI, de la Maza, L. M. and E. M. Peterson, eds. (Elsevier, New York, 1987) 331-351.

Cooper, G. M., "Cellular Transforming Genes", Science 217: 801-806 (1982).

Couch, R. B., "Rhinoviruses", Virology, Second Edition, edited by B. N. Fields, D. M. Knipe et al. Raven Press, Ltd., New York, 607-629 (1990).

Creighton, T. E., Proteins by W. H. Freeman and Company, New York, 33-34 (1984).

Cunningham et al., "Neural Cell Adhesion Molecule: Structure, Immunoglobulin-Like Domains, Cell Surface Modulation, and Alternative RNA Splicing", Science 236:799-806 (1987).

Cybulsky, M. I. and Gimbrone, Jr., M. A., "Endothelial Expression of a Mononuclear Leukocyte Adhesion Molecule During Atherogenesis", Science 251:788-791 (1991).

Deen et al., "A Soluble Form of CD4 (T4) Protein Inhibits AIDS Virus Infection", Nature 331:82-86 (1988).

Dick, E. C., "Experimental Infection of Chimpanzees with Human Rhinovirus Types 14 and 43", Proceedings Of The Society For Experimental Biology And Medicine 127:1079-1081 (1968).

Douglas et al., "Prophylactic Efficacy of Intranasal Alpha 2-Interferon Against Rhinovirus Infections in the Family Setting", The New England J. of Med. 314:65-70 (1986).

Dustin et al., "Induction by IL 1 and Interferon-.gamma.: Tissue Distribution, Biochemistry, and Function of a Natural Adherence Molecule (ICAM-1)", J. Immunol. 137(1):245-254 (1986).

Dustin et al., "Supergene Families Meet in the Immune System" Immunology Today, 9(7 and 8):213-215 (1988).

Dustin et al., "Correlation of CD2 Binding and Functional Properties of Multimeric and Monomeric Lymphocyte Function-Associated Antigen 3", J. Exp. Med. 169:503-517 (1989).

Ey, P. L., et. al., "Isolation of Pure IgG1, IgG2a, and IgG2b Immunoglobulins from Mouse Serum Using Protein A--Sepharose", Immunochemistry 15:429-436 (1978).

Fisher et al., "HIV Infection is Blocked in vitro by Recombinant Soluble CD4", Nature 331:76-78 (1988).

Fox et al., "Prevention of a Rhinovirus and Poliovirus Uncoating by WIN 51711, a New Antiviral Drug", Antimicrob. Ag. and Chemotherapy 30: 110-116 (1986).

Galfrey et. al., "Antibodies to Major Histocompatibility Antigens Produced by Hybrid Cell Lines", Nature 266:550-552 (1977).

Gething, M. J. and Sambrook, J., "Construction of Influenza Haemagglutinin Genes that Code for Intracellular and Secreted Forms of the Protein" Nature 300:598-603 (1982).

Ginsberg et al., "Inhibition of Fibronectin Binding to Platelets by Proteolytic Fragments and Synthetic Peptides Which Support Fibroblast Adhesion", J. Biol. Chem. 260(7):3931-3936 (1985).

Giranda et al., "Modeling of the Human Intercellular Adhesion Molecule-1, the Human Rhinovirus Major Group Receptor" Proteins: Structure, Function, and Genetics, 7:227-233 (1990).

Gough, N., "Putting A Stop To An Immunoglobulin Message", Trends Genet. 3(9):238-240 (1987).

Gower et al., "Alternative Splicing Generates a Secreted Form of N-CAM in Muscle and Brain", Cell 55:955-964 (1988).

Graham, F. L., and Van der Eb, A. J., "A New Technique for the Assay of Infectivity of Human Adenovirus 5 DNA", Virology 52:456-467 (1973).

Green et al., "Immunogenic Structure of the Influenza Virus Hemagglutinin", Cell 28:477-487 (1982).

Greve et al., "The Major Human Rhinovirus Receptor Is ICAM-1", Cell 56:839-847 (1989).

Gross-Bellard et al., "Isolation of High-Molecular-Weight DNA from Mammalian Cells", Eur. J. Biochem. 36:32-38 (1973).

G ussow, D. and Ploegh, H., "Soluble class I antigens: a conundrum with no solution?", Immunology Today 8(7,8):220-222 (1987).

Hamparian et al., "A Collaborative Report: Rhinoviruses-Extension of the Numbering System from 89 to 100", Virology 159:191-192 (1987).

Hayden et al., "Safety and Efficacy of Intranasal Pirodavir (R77975) in Experimental Rhinovirus Infection", Antimicrob. Agents Chemother. 36(4): 727-732 (1992).

Hayden et al., "Prevention of Natural Colds by Contact Prophylaxis with Intrnasal Alpha2-Interferon", The New England Journal of Medicine, 314(2):71-75 (1986).

Holland, J. J. and McLaren, L. C., "The mammalian cell-virus relationship. II. Absorption, Reception and Eclipse of Poliovirus by HeLa Cells" J. Exp. Med. 109:487-504 (1959).

Holland, J. J., "Receptor affinities as Major Determinants of Enterovirus Tissue Tropisms in Humans", Virology 15:312-326.

Horton et al., "Gene Splicing by Overlap Extension: Tailor-Made Genes Using the Polyerase Chain Reaction", BioTechniques 8(5):528-535 (1990).

Hussey et. al., "A Soluble CD4 Protein Selectively Inhibits HIV Replication and Syncytium Formation" Nature, 331:78-81 (1988).

Kamarck, M. E., and Ruddle, F. H., "Somatic Cell Genetics and the Human Gene Map", Chapter 105 in Handbook of Experimental Immunology in Four Volumes, vol. 3: Genetics and Molecular Immunology, D. M. Weir, ed. (Blackwell Scientific Publications, Boston, MA, 1986).

Katz et al., "Chromosome Mapping of Cell Membrane Antigens Expressed on Activiated B Cells", Eur. J. Immunol., 15:103-106 (1985).

Kavenoff, R., and Zimm, B. H., "Chromosome-Sized DNA Molecules from Drosophila", Chromosoma (Berl.) 41:1-27 (1973).

K uhn et al., "Gene Transfer, Expression, and Molecular Cloning of the Human Transferrin Receptor Gene", Cell 37:95-103 (1984).

Lebman et al., "A Monoclonal Antibody that Detects Expression of Transferrin Receptor in Human Erythroid Precursor Cells", Blood 59(3): 671-678 (1982).

Littlefield, J. W., "Selection of Hybrids from Matings of Fibroblasts in vitro and Their Presumed Recombinants", Science 145:709-710 (1964).

Lonberg-Holm et al., "Unrelated Animal Viruses Share Receptors", Nature 259:679-681 (1976).

Margulies, D. H., et. al., "Engineering Soluble Major Histocompatibility Molecules: Why and How", Immunol. Res. 6: 101-116 (1987).

Marlin, S. D. and Springer, T. A., "Purified Intercellular Adhesion Molecule-1 (ICAM-1) Is a Ligand for Lymphocyte Function-Associated Antigen 1 (LFA-1)", Cell 51:813-819 (1987).

Marlin et al., "A Soluble Form of Intercellular Adhesion Molecule-1 Inhibits Rhinovirus Infection", Nature 344:70-72 (1990).

Marsh et al., "Antibody-toxin Conjugation", Immunotoxins by Kluwer Academic Publishers, Boston, Dordrecht, Lancaster 213-237 (1988).

McClelland et al., "Identification of Monoclonal Antibody Epitopes and Critical Residues for Rhinovirus in Domain 1 of ICAM-1", PNAS 88(18): 7993-7997 (1991).

McClelland et al., "Transfectant cell lines which express the major human rhinovirus receptor, their preparation, and their uses", Immuno. 112: 117179 (1990).

McCray, J. and Werner, G., "Different Rhinovirus Serotypes Neutralized by Antipeptide Antibodies", Nature 329:736-738 (1987).

Medical Microbiology: "An Introduction to Infectious Diseases", 2nd ed., J. C. Sherris, ed. (Elsevier Science Publishing Co., Inc., N.Y. 1990) pp. 514-515.

Medrano, L. and Green, H., "Picornavirus Receptors and Picornavirus Multiplication in Human-Mouse Hybrid Cell Lines", Virology 54:515-524 (1973).

Melchers et al., Lymphocyte Hybridomas, vol. 81 of Current Topics in Microbiology and Immunology, W. Arber, W. Henle, P. H. Hofschneider, J. H. Humphrey, J. Klein, P. Koldovsky, H. Koprowski, O. Maaloe, F. Melchers, R. Rott, H. G. Schweiger, L. Syrucek, P. K. Vogt, eds (Springer Verlang, New York, 1978).

Mendelsohn et al., "Transformation of a Human Poliovirus Receptor Gene into Mouse Cells", PNAS 83:7845-7849 (1986).

Minor, P. D., "Growth, Assay and Purification of Picornaviruses", in Virology: A Practical Approach, B. W. J. Mahy, ed. (IRL Press Limited, Oxford, England), 25-41 (1985).

Minor et al., "Monoclonal antibodies which block cellular receptors of poliovirus", Virus Research 1:203-212 (1984).

Niman et al., "Anti-peptide antibodies detect oncogene-related proteins in urine", PNAS 82:7924-7928 (1985).

Nobis et al., "Production of a Monoclonal Antibody against an Epitope on HeLa Cells that Is the Functional Poliovirus Binding Site", J. Gen. Virol. 66:2563-2569 (1985).

Parham, P., "Monoclonal Antibodies Against HLA Products and Their use in Immunaffinity Purification," Methods in Enzymology 92:110-138 (1983).

Pepinsky et al., "The Increased Potency of Crossed-linked Lymphocyte Function-associated Antigen-3 (LFA-3) Multimers Is a Direct Consequence of Changes in Valency", J. Biol. Chem. 266(27):18244-18249 (1991).

Peterson, A. and Seed, B., "Genetic Analysis of Monoclonal Antibody and HIV Binding Sites on the Human Lymophocyte Antigen CD4", Cell 54:65-72 (1988).

Rossman et al., "Structure of a Human Common Cold Virus and Functional Relationship to other Picornaviruses", Nature 317:145-153 (1985).

Rothlein et al., "A Form of Circulating ICAM-1 In Human Serum", J. Immuno. 147(11):3788-3793 (1991).

Rothlein et al., "A Human Intercellular Adhesion Molecule (ICAM-1) Distinct From LFA-1", J. Immuno. 137(4):1270-1274 (1986).

Ruddle et al., "DNA-Mediated Gene Transfer in Mammalian Gene Cloning", Genetic Engineering 6:319-338 (1984).

Ruoslahti et al., "Synthetic Peptides in the Analysis of Cell Adhesion," in Synthetic Peptides in Biology and Medicine Elsvier Science Publishers, pp. 191-197 (1985).

Saiki et al., "Enzymatic Amplification of .beta.-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia," Science 230:1350-1354 (1985).

Sambrook et al., Molecular Cloning: A Laboratory Manual, 2d ed. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989) pp. 1.21-1.52.

Scopes, R. K., "Separation By Precipitation," in Protein Purification: Principles & Practice (1982) Springler Verlag, NY, pp. 39-46.

Seed, B. and Aruffo, A., "Molecular Cloning of the CD2 antigen, the T-Cell Erythrocyte Receptor, by a Rapid Immunoselection Procedure," PNAS 84:3365-3369 (1987).

Seed, B., "An LFA-3 cDNA Encodes a Phospholipid-Linked Membrane Protein Homologous to its Receptor CD2," Nature 329:840-842 (1987).

Seth et al., "Circulating ICAM-1 isoforms: Diagnostic Prospects for Inflammatory and Immune Disorders," Lancet 338:83-84 (1991).

Sherman-Gold, R., "Companies Pursue Therapies Based on Complex Cell Adhesion Molecules", Genetic Engineering News pp. 6-7, 14 (Jul. 1993).

Sherry, B. and Rueckert, R., "Evidence for at Least Two Dominant Neutralization Antigens on Human Rhinovirus 14," J. Virol. 53(1): 137-143 (1985).

Shih, C. and Weinberg, R. A., "Isolation of a Transforming Sequence from a Human Bladder Carcinoma Cell Line," Cell 29:161-169 (1982).

Siddique et al., "The Poliovirus Sensitivity (PVS) Gene Is on Chromosome 19q12->q13.2", Genomics 3:156-160 (1988).

Simmons et al., "ICAM, an Adhesion Ligand of LFA-1, is Homologous to the Neural Cell Adhesion Molecule NCAM," Nature 331:624-627 (1988).

Skerra, A. and Pluckthun, A., "Assembly of a Functional Immunoglobulin FV Fragment in Escherichia Coli" Science, 240:1038-1041 (1988).

Smith, T. J., et. al., "The Site of Attachment in Human Rhinovirus 14 for 4 Antiviral Agents that Inhibit Uncoating", Science 233:1286-1293 (1986).

Smith et al., "Blocking of HIV-1 Infectivity by a Soluble, Secreted Form of the CD4 Antigen," Science 238:1704-1707 (1987).

Smith et al., "Modification and Secretion of Human Interleukin 2 Produced in Insect Cells by a Baculovirus Expression Vector", PNAS 82:8404-8408 (1985).

Springer, T. A., "Adhesion Receptors of the Immune System", Nature 346:425-434 (1990).

Staunton et al., "Primary Structure of ICAM-1 Demonstrates Interaction between Members of the Immunoglobulin and Integrin Supergene Families," Cell 52:925-933 (1988).

Staunton et al., "The Arrangement of the Immunoglobulin-Like Domains of ICAM-1 and the Binding Sites for LFA-1 and Rhinovirus," Cell 61:243-254 (1990).

Staunton et al., "A Cell Adhesion Molecule, ICAM-1, is the Major Surface Receptor for Rhinoviruses," Cell 56:849-853 (1989).

Tomassini, J. E., "Isolation, Characterization and Cloning of the Cellular Receptor for the Major Group of Human Rhinoviruses," Ph.D. Thesis, University of Pennsylvania (1986).

Tomassini, J. E. and Colonno, R. J., "Isolation of a Receptor Protein Involved in Attachment of Human Rhinoviruses," J. Virol. 58(2): 290-295 (1986).

Tomassini et al., "cDNA Cloning Reveals that the Major Group Rhinovirus Receptor on HeLa Cells is Intercellular Adhesion Molecule 1," PNAS 86:4907-4911 (1989).

Towbin et. al., "Electrophoretic Transfer of Proteins from Polyacrylamide Gels to Nitrocellulose Sheets: Procedure and Some Applications", PNAS 76(9):4350-4354 (1979).

Traunecker et al., "Highly Efficient Neutralization of HIV with Recombinant CD4-Immunoglobulin Molecules," Nature 339:68-70 (1989).

Urlaub, G. and Chasin, L. A., "Isolation of Chinese Hamster Cell Mutants Deficient in Dihydrofolate Reductase Activity," PNAS USA 77(7): 4216-4220 (1980).

Wade, N., "Hybridomas: A Potent New Biotechnology," Science 208: 692-693 (1980).

Welsh, K. I., "Antibody Production Made Easier," Nature 266: 495 (1977).

Wigler et al., "Transformation of Mammalian Cells with Genes from Procaryotes and Eucaryotes," Cell 16:777-785 (1979).

Williams, A. F., "A Year in the Life of the Immunoglobulin Superfamily", Immunology Today 8(10):298-303 (1987).

Williams, A. F. and Barclay, A. N., "The Immunoglobulin Superfamily-Domains for Cell Surface Recognition.sup.1,2 ", Ann. Rev. Immunol. 6:381-405 (1988).

Winther et al., "Sites of Rhinovirus Recovery After Point Inoculation of the Upper Airway", JAMA 256(13):1763-1767 (1986).

Zettlmeissl et al., "Expression and Characterization of Human CD4: Immunoglobulin Fusion Proteins," DNA and Cell Biology 9:347-353 (1990).