WO2001016182A2 - POLYPEPTIDES THAT BIND HIV gp120 AND RELATED NUCLEIC ACIDS, ANTIBODIES, COMPOSITIONS, AND METHODS OF USE - Google Patents

Abstract

The present invention provides, among other things, a polypeptide that binds with the gp120 envelope protein of HIV, in particular HIV-1, under physiological conditions, a nucleic acid that encodes such a polypeptide and can be expressed in a cell, a composition comprising such a polypeptide or nucleic acid or an antibody and a carrier therefor, a composition comprising a solid support matrix to which is attached an above-described polypeptide or an anti-antibody to a specified polypeptide sequence, a method of making an antibody to gp120, and a method of removing HIV from a bodily fluid.

Description

POLYPEPTIDES THAT BIND HIV gpl20 AND RELATED NUCLEIC ACIDS, ANTIBODIES, COMPOSITIONS, AND METHODS OP USE

TECHNICAL FIELD OF THE INVENTION The present invention relates to polypeptides with homology to regions of domains of the human chemokine receptors CCR5, CXCR4, and STRL33, as well as domains of CD4 that bind with human immunodeficiency virus (HIV) , • in particular HIV-1 glycoprotein 120 (gpl20) envelope protein. The present invention also relates to nucleic acids encoding such polypeptides, antibodies, compositions comprising such polypeptides, nucleic acids or antibodies, and methods of using the same.

BACKGROUND OF THE INVENTION

There are seven transmembrane chemokine receptors that act as cofactors for HIV infection. The cof ctors enable entry of HIV-1 into CD4^* T cells and macrophages (Premack et al., Nature Medicine 2: 1174-78 (1996); and Zhang et al., Nature 383 : 768 (1996)).

The presence of chemokines has an inhibitory effect on HIV-1 attachment to, and infection of, susceptible cells. Additionally, some mutations in chemokine receptors have been shown to result in resistance to HIV-1 infection. For example, a 32-nucleotide deletion within the CCR5 gene has been described in subjects who remained uninfected despite repeated exposures to HIV-1 (Huang et al., Nature Medicine 2: 1240-43 (1996)).

Evidence also exists for the physical association of a ternary complex between chemokine receptors, CD4, and HIV-1 gpl20 envelope glycoprotein on cell membranes (Lapham et al., Science 274: 602-05 (1996)). Receptor signaling and cell activation are probably not required for the anti-HIV-1 effect of chemokines since a RANTES analog lacking the first eight amino-terminal amino acids, RANTES (9-68), lacked chemotactiσ and leukocyte- activating properties, but bound to multiple chemokine receptors and inhibited infection by macrophage-tropic HIV-1 (Arenzana-Seladedos et al., Nature 383: 400 (1996)). Cumulatively, the above described results suggest that the interaction between gpl20, CD4, and at least one chemokine receptor is obligatory for HIV-1 infection. Accordingly, reagents that interfere with the binding of gpl20 to chemokine receptors and to CD4 are used in the biological and medical arts. However, there presently exists a need for additional reagents that can compete with one or more proteins of the gpl20-CD4-chemokine receptor complex to assist in basic biological or viral research, and to assist in medical intervention in the HIV-1 pandemic. It is an object of the present invention to provide such reagents. This and other objects and advantages, including additional inventive features, will be apparent from the description provided herein.

BRIEF SUMMARY OF THE INVENTION

The present invention provides a polypeptide that binds with HIV gpl20 under physiological conditions. Multiple embodiments of the present inventive polypeptide are provided, and each embodiment possesses a degree of homology to at least one of the human CCR5, CXCR4 and STRL33 chemokine receptors, and the human CD4 cell- surface protein.

In a first embodiment, the present invention provides a polypeptide comprising the amino acid sequence YDIXYYXXE, wherein X is any synthetic or naturally occurring amino acid residue, and the polypeptide comprises less than about 100 contiguous amino acids that are identical to, or, in the alternative, substantially identical to, the amino acid sequence of the human CCR5 chemokine receptor. A preferred polypeptide of this first embodiment comprises the amino acid sequence YDIN*YYT*S*E. A more preferred polypeptide of this first embodiment comprises the amino acid sequence YDINYYTSE, wherein each letter is the standard one-letter abbreviation for an amino acid residue (i.e., for example, N denotes asparaginyl, T denotyes threoninyl, and S denotes serinyl) . The polypeptide of the first embodiment can comprise the amino acid sequence M*D*YQ*V*S*SP*IYDIN*YYT*S*E. Preferably, the polypeptide comprises the amino acid sequence MDYQVSSPIYDINYYTSE. In a second embodiment, the present invention provides a polypeptide comprising the amino acid sequence XEXIXIYXXXNYXXX, wherein X is any synthetic or naturally occurring amino acid and wherein said polypeptide comprises less than about 100 contiguous amino acid that are identical to or substantially identical to the amino acid sequence of the human CXCR4 chemokine receptor. The polypeptide can consist essentially of, or consist of, the sequence EXIXIYXXXNY. Preferably, the polypeptide comprises the sequence M*EG*IS*IYT*S*D*NYT*E*E*. Preferably, M*EG*IS*IYT*S*D*NYT*E*E* is M*EGISIYTSDNYT*E*E* .

In a third embodiment, the present invention provides a polypeptide comprising the amino acid sequence EHQAFLQFS, wherein said polypeptide comprises less than about 100 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human STRL33 chemokine receptor. The polypeptide can consist essentially of, or consist of, the sequence EHQAFLQFS.

In a fourth embodiment, the present invention provides a polypeptide comprising at least a portion of an amino acid sequence selected from the group consisting of LPPLYSLVFIFGFVGNML, QWDFGNTMCQLLTGLYFIGFFS, SQYQFWKNFQTLKIVILG, APYNIVLLLNTFQEFFGLNNCS, and

YAFVGEKFRNYLLVFFQK, wherein said polypeptide comprises less than about 100 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human CCR5 chemokine receptor. In a fifth embodiment, the present invention provides a polypeptide comprising at least a portion of an amino acid sequence selected from the group consisting of LL TIPDFIFANVSEADD, WFQFQHIMVGLILPGIV, and IDSFILLEIIKQGCEFEN, wherein said polypeptide comprises less than about 100 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human CXCR4 chemokine receptor.

In a sixth embodiment, the present invention provides a polypeptide comprising at least a portion of an amino acid sequence selected from the group consisting of LVISIFYHKLQSLTDVFL, PFAYAGIHE VFGQVMC, EAISTWLATQMTLGFFL, LTMIVCYSVIIKTLLHAG, MAVFLLTQMPFNLMKFIRSTHW, HWEYYAMTSFHYTIMVTE, ACLNPVLYAFVSLKFRKN and SKTFSASHNVEATSMFQL, wherein said polypeptide comprises less than about 100 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human STRL33 chemokine receptor.

In a seventh embodiment, the present invention provides a polypeptide comprising at least a portion of an amino acid sequence selected from the group consisting of DTYICEVED, EEVQLLVFGLTANSD, THLLQGQSLTLTLES, and GEQVEFSFPLAFTVE, wherein said polypeptide comprises less than about 100 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human CD4 cell-surface protein.

In the fourth to seventh embodiments, any selected portion of the polypeptide can comprise from 1 to about 6 conservative amino acid substitutions. In an alternative, the polypeptide can be partially defined by an absence of a polypeptide sequence, outside the region of the portion selected from the foregoing sequences, that has five, or ten, contiguous amino acid residues that have a sequence that consists of an amino acid sequence that is identical to or substantially identical to the protein to which the polypeptide has homology (i.e., CCR5, CXCR4, STRL33, or CD4) . In yet another alternative, the polypeptide can lack a sequence of five or ten contiguous amino acids which are identical to or substantially identical to the sequence of the protein with which the sequence has homology except that one or more conservatively or neutrally substituted amino acids replace part of the sequence of the protein to which the polypeptide has homology. Additionally, any embodiment of the present inventive polypeptide can also comprise a pharmaceutically acceptable substituent. Any embodiment of the present inventive polypeptide can be incorporated into a composition, which further comprises a carrier. Any suitable embodiment of the present inventive polypeptide can be encoded by a nucleic acid that can be expressed in a cell. In this regard, the present invention further provides a vector comprising such a nucleic acid. The nucleic acids and vectors also can be incorporated into a composition comprising a carrier.

Additionally, the present invention provides a method of making an antibody to a polypeptide of the present invention. The present invention also provides a method of prophylactically or therapeutically treating an HIV infection in a mammal.

Additionally, the present invention provides an anti-idiotypic antibody comprising an internal image of a portion of gpl20, as well as a method of selecting such an antibody.

The present invention also provides a method of making an antibody to a portion of the gpl20 protein that binds with a portion of CCR5, CXCR4, STRL33, or CD4, as well as the immunizing compound used to make the antibody, and the antibody itself. In another embodiment of the present invention, a method of removing HIV-1 from a bodily fluid is provided. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 depicts a listing of synthetic amino acids available (from Bachem, King of Prussia, PA) for incorporation into polypeptides of the present invention.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a polypeptide that binds with gpl20 of HIV, in particular HIV-1, more particularly HIV-l^, under physiological conditions. The polypeptide has a number of uses including, but not limited to, the use of the polypeptide to elucidate the mechanism by which HIV, such as HIV-1, attaches to and/br infects a particular cell, to induce an immune response in a mammal, in particular a human, to HIV, in particular HIV-1, and to inhibit the replication of HIV, in particular HIV-1, in an infected mammal, in particular a human.

Multiple embodiments of the present inventive polypeptide are provided. Each embodiment of the polypeptide has a degree of homology to at least one of the human CCR5, CXCR4. and STRL33 chemokine receptors, or the human CD4 cell-surface protein. In each embodiment provided herein, a letter indicates the standard amino acid designated by that letter, and a letter followed directly by an asterisk (*) preferably represents the amino acid represented by the letter (e.g., N represents asparaginyl and T represents threoninyl) , or a synthetic or naturally occurring conservative or neutral substitution therefor. Additionally, in accordance with convention, all amino acid sequences provided herein are given either from left to right, or top to bottom, such that the first amino acid is amino-terminal and the last is carboxyl-terminal. The synthesis of polypeptides, either synthetically (i.e., chemically) or biologically, is within the skill in the art. It is within the skill of the ordinary artisan to select synthetic and naturally occurring amino acids that make conservative or neutral substitutions for any particular naturally occurring amino acids. The skilled artisan desirably will consider the context in which any particular amino acid substitution is made, in addition to considering the hydrophobicity or polarity of the side-chain, the general size of the side chain, and the pK value of side-chains with acidic or basic character under physiological conditions. For example, lysine, arginine, and histidine are often suitably substituted for each other, and more often arginine and lysine. As is known in the art, this is because all three amino acids have basic side chains, whereas the pK value for the side-chains of lysine and arginine are much closer to each other (about 10 and 12) than to histidine (about 6) . Similarly, glycine, alanine, valine, leucine, and isoleucine are often suitably substituted for each other, with the proviso that glycine is frequently not suitably substituted for the other members of the group. This is because each of these amino acids are relatively hydrophobic when incorporated into a polypeptide, but glycine 's lack of an α-carbon allows the phi and psi angles of rotation (around the α-carbon) so much conformational freedom that glycinyl residues can trigger changes in conformation or secondary structure that do not often occur when the other amino acids are substituted for each other. Other groups of amino acids frequently suitably substituted for each other include, but are not limited to, the group consisting of glutamic and aspartic acids; the group consisting of phenylalanine, tyrosine and tryptophan; and the group consisting of serine, threonine and, optionally, tyrosine. Additionally, the skilled artisan can readily group synthetic amino acids with naturally occurring amino acids. In the context of the present invention, a polypeptide is "substantially identical" to another polypeptide if it comprises at least about 80% identical amino acids. Desirably, at least about 50% of the non-identical amino acids are conservative or neutral substitutions. Also, desirably, the polypeptides differ in length (i.e., due to deletion mutations) by no more than about 10%.

In a first embodiment, the present invention provides a polypeptide comprising the amino acid sequence YDIXYYXXE, wherein X is any synthetic or naturally occurring amino acid residue, and the polypeptide comprises less than about 100 contiguous amino acids, preferably less than about 50 amino acids, more preferably less than about 25 amino acids, and yet more preferably less than about 13 amino acids that are identical to, or, in the alternative, substantially identical to, the amino acid sequence of the human CCR5 chemokine receptor.

Preferably, the polypeptide of the first embodiment comprises YDIXYYXXE, wherein the amino moiety of the amino-terminal tyrosinyl residue is not bound to another amino acid residue via a peptidic bond, and the carboxyl moiety of the glutamyl residue is not bound to another amino acid residue via a peptidic bond. However, the polypeptide can consist essentially of YDIXYYXXE and, optionally, can be modified by one or more pharmaceutically acceptable substituents, such as, for example, t-boc or a saccharide.

More particularly, the polypeptide comprises the amino acid sequence YDIN*YYT*S*E. Preferably, N* is asparaginyl, T* is threoninyl, and S* is serinyl.

The polypeptide of the first embodiment can comprise a dodecapeptide selected from the amino acid sequence M*D*YQ*V*S*SP*IYDIN*YYT*S*E. More preferably, the polypeptide of the first embodiment comprises the amino acid sequence MDYQVSSPIYDINYYTSE.

In a second embodiment, the present invention provides a polypeptide comprising the amino acid sequence XEXIXIYXXXNYXXX_/ wherein X is any synthetic or naturally occurring amino acid, and the polypeptide comprises less than about 100 contiguous amino acids,, preferably less than about 50 amino acids, and more preferably less than about 25 amino acids, that are identical to or substantially identical to the amino acid sequence of the human CXCR4 chemokine receptor. Optionally, the polypeptide consists essentially of, or consists of, the sequence EXIXIYXXXNY.

In a preferred polypeptide of this second embodiment, the polypeptide comprises the amino acid sequence M*EG*IS*IYT*S*D*NYT*E*E*. Preferably, M*EG*IS*IYT*S*D*NYT*E*E* is M*EGISIYTSDNYT*E*E* . In a third embodiment, the present invention provides a polypeptide comprising the amino acid sequence EH^QAFLQFS, wherein the polypeptide comprises less than about 100 contiguous amino acid residues, preferably less than about 50 contiguous amino acid residues, more preferably less than about 25 contiguous amino acid residues, that are identical to or substantially identical to the amino acid sequence of the human STRL33 chemokine receptor. The polypeptide can consist essentially of, or consist of, the sequence EHQAFLQFS.

The first three embodiments of the present invention provide, among other things, polypeptides having substantial identity or identity to the amino-terminal regions of the chemokine receptors CCR5, CXCR4, and STRL33. These first three embodiments form a first group of embodiments of the present invention. The present invention also provides, in a second group of embodiments, polypeptides having substantial identity or identity to an internal region of the human chemokine receptors CCR5, CXCR4, and STRL33, as well as to the leukocyte cell-surface protein CD4.

This second group of embodiments provides a polypeptide that binds with HIV gpl20 under physiological conditions and comprises at least a portion of or .all of an amino acid sequence selected from the group consisting of LPPLYSLVFIFGFVGNML, QWDFGNTMCQLLTGLYFIGFFS, SQYQFWKNFQTLKIVILG, APYNIVLLLNTFQEFFGLNNCS, and YAFVGEKFRNYLLVFFQK, wherein the polypeptide comprises less than about 100 amino acids that are identical to or substantially identical to the amino acid sequence of the human CCR5 chemokine receptor; or selected from the group consisting of LLLTIPDFIFANVSEADD (165-182) , WFQFQHIMVGLILPGIV (197-214) , and IDSFILLEIIKQGCEFEN (261-278) , wherein the polypeptide comprises less than about 100 amino acids that are identical to or substantially identical to the amino acid sequence of the human CXCR4 chemokine receptor; or selected from the group consisting of LVISIFYHKLQSLTDVFL (53-70), PFWAYAGIHEWVFGQVMC (85-102), EAISTWLATQMTLGFFL (185-202) , LTMIVCYSVIIKTLLHAG (205- 222) , MAVFLLTQMPFNLMKFIRSTHW (237-258) ,

HWEYYAMTSFHYTIMVTE (257-274) , ACLNPVLYAFVSLKFRKN (281- 298) and SKTFSASHNVEATSMFQL (325-342) , wherein the polypeptide comprises less than about 100 amino acids that are identical to a substantially identical to the amino acid sequence of the human STRL33 chemokine receptor; or selected from the group consisting of DTYICEVED, EEVQLLVFGLTANSD, THLLQGQSLTLTLES, and GEQVEFSFPLAFTVE, wherein the polypeptide binds with HIV gpl20 under physiological conditions and comprises less than about 100 amino acids that are identical to or substantially identical to the amino acid sequence of the human CD4 cell-surface protein. Optionally, the recited amino acid sequences can comprise 1 to about 6 conservative or neutral amino acid substitutions.

The polypeptides of this second group of embodiments preferably comprise less than about 50 amino acid residues, and more preferably less than about 25 amino acid residues, and yet more preferably no additional amino acid residues, that are identical to a protein that naturally has the recited amino acid sequence. The polypeptide can be alternatively characterized by an absence of a region, outside the above-recited amino acid sequences, that has about five, or about ten, contiguous amino acid residues that have a sequence that consists of an amino identical and conservatively substituted residues as an amino acid sequence of the protein to which the polypeptide of the compound has homology.

Any embodiment of the present inventive polypeptide can also comprise a pharmaceutically acceptable substituent, attachment of which is within the skill in the art. The pharmaceutically acceptability of substituents are understood by those skilled in the art. For example, a pharmaceutically acceptable substituent can be a biopolymer, such as a polypeptide, an RNA, a DNA, or a polysaccharide. Suitable polypeptides comprise fusion proteins, an antibody or fragment thereof, a cell adhesion molecule or a fragment thereof, or a peptide hormone. Suitable polysaccharides comprise polyglucose moieties, such as starch and their derivatives, such as heparin. The pharmaceutically acceptable substituent also can be any suitable lipid or lipid-containing moiety, such as a lipid of a liposome or a vesicle, or even a lipophilic moiety, such as a prostaglandin, a steroid hormone, or a derivative thereof. Additionally, the pharmaceutically acceptable substituent can be a nucleotide or nucleoside, such as nicotine adenine dinucleotide or thymine, an amino acid residue, a saccharide or disaccharide, or the residue of another biomolecule naturally occurring in a cell, such as inositol, a vitamin, such as vitamin C, thiamine, or nicotinic acid. Synthetic organic moieties also can be pharmaceutically acceptable substituents, such as t-butyl carbonyl, an acetyl moiety, quinine, polystyrene and other biologically acceptable polymers. Optionally, a pharmaceutically acceptable substituent can be selected from the group consisting of a C_J-C_J,, alkyl, a C₂-C₁₈ alkenyl, a Ca-C_ιa alkynyl, a C₃-C_lβ aryl, a C₇-C_lβ alkaryl, a C₇-C₁₈ aralkyl, and a C₃-C_lβ cycloalkyl, wherein any of the foregoing moieties that are cyclic comprise from 0 to 2 atoms per carbocydic ring, which can be the same or different, and are selected from the group consisting of nitrogen, oxygen, and sulfur.

Any of the substituents from this group can be substituted by one to six substituent moieties, which can be the same or different, selected from the group consisting of an amino moiety, a carbamate moiety, a carbonate moiety, hydroxyl, a phosphamate moiety, a phosphate moiety, a phosphonate moiety, a pyrophosphate moiety, a triphosphate moiety, a sulfamate moiety, a sulfate moiety, a sulfonate moiety, a C₁-C₈ monoalkylamine moiety, a

dialkylamine moiety, and a C_x-C_a trialkylamine moiety.

Any embodiment of the present inventive polypeptide can be encoded by a nucleic acid and can be expressed in a cell. The skilled artisan will recognize that the encoded polypeptide as well as any pharmaceutically acceptable substituent to be incorporated into the polypeptide, e.g., a formyl or acetyl substituent on an amino-terminal methionine or a saccharide, will preferably be produced by a cell that can express the polypeptide of the present invention. Accordingly, the amino acids incorporated into the polypeptide encoded by the nucleic acid are preferably naturally occurring.

A nucleic acid as described above can be cloned into any suitable vector and can be used to transduce, transform, or transfect any suitable host. The selection of vectors and methods to construct them are commonly known to persons of ordinary skill in the art and are described in general technical references (see, in general, "Recombinant DNA Part D," Methods in Enzymology, Vol. 153, Wu and Grossman, eds., Academic Press (1987)). Desirably, the vector comprises regulatory sequences, such as transcription and translation initiation and termination codons, which are specific to the type of host (e.g., bacterium, fungus, plant, or animal) into which the vector is to be inserted, as appropriate and taking into consideration whether the vector is DNA or RNA. Preferably, the vector comprises regulatory sequences that are specific to the genus of the host. Most preferably, the vector comprises regulatory sequences that are specific to the species of the host and is optimized for the expression of an above-described polypeptide.

Constructs of vectors, which are circular or linear, can be prepared to contain an entire nucleic acid sequence as described above or a portion thereof ligated to a replication system that is functional in a prokaryotic or eukaryotic host cell. Replication systems can be derived from ColEl, 2 mμ plasmid, λ, SV40, bovine papilloma virus, and the like. Suitable vectors include those designed for propagation and expansion, or for expression, or both. A preferred cloning vector is selected from the group consisting of the pUC series, the pBluescript series (Stratagene, LaJolla, CA) , the pET series (Novagen, Madison, WI) , the pGEX series (Pharmacia Biotech, Uppsala, Sweden) , and the pEX series (Clonetech, Palo

Alto, CA) . Examples of animal expression vectors include pEUK-Cl, pMAM and pMAMneo (Clonetech, Palo Alto, CA) .

An expression vector can comprise a native or normative promoter operably linked to a nucleic acid molecule encoding an above-described polypeptide. The selection of promoters, e.g., strong, weak, inducible, tissue-specific and developmental-specific, is within the skill in the art. Similarly, the combining of a nucleic acid molecule as described above with a promoter is also within the skill in the art.

The skilled artisan will also recognize that the polypeptide has ability to bind th gpl20 protein, which is most often found outside of cells. Accordingly, the present inventive nucleic acid advantageously can comprise a nucleic acid sequence that encodes a signal sequence such that a signal sequence is .translated as a fusion protein with the polypeptide of the present inventive polypeptide to form a signal sequence- polypeptide fusion. The signal sequence can cause secretion of the entire polypeptide, including the signal sequence (which is a pharmaceutically acceptable substituent) , or can be cleaved from the polypeptide (i.e., the polypeptide of the compound) prior to, or during, secretion so that at least the present inventive polypeptide is secreted out of a cell in which the nucleic acid is expressed. Alternatively, the nucleic acid comprises or encodes an antisense nucleic acid molecule or a ribozyme that is specific for a specified amino acid sequence of an above- described polypeptide . A nucleic acid sequence introduced in antisense suppression generally is substantially identical to at least a portion of the endogenous gene or gene to be repressed, but need not be identical. Thus, the vectors can be designed such that the inhibitory effect applies to other proteins within a family of genes exhibiting homology or substantial homology to the target gene. The introduced sequence also need not be full-length relative to either of the primary transcription product or the fully processed mRNA. Generally, higher homology can be used to compensate for the use of a shorter sequence.

Furthermore, the introduced sequence need not have the same intron or exon pattern, and homology of non-coding segments will be equally effective.

Ribozymes also have been reported to have use as a means to inhibit expression of endogenous genes. It is possible to design ribozymes that specifically pair with virtually any target RNA and cleave the phosphodiester backbone at a specific location, thereby functionally inactivating the target RNA. In carrying out this cleavage, the ribozyme is not itself altered and is, thus, capable of recycling and cleaving other molecules, making it a true enzyme. The inclusion of ribozyme sequences within antisense RNAs confers RNA-cleaving activity upon them, thereby increasing the activity of the constructs. The design and use of target RNA- specific ribozymes is described in Haseloff et al., Nature 334: 585-591 (1988).

Further provided by the present invention is a composition comprising an above-described polypeptide or nucleic acid and a carrier therefor. Another composition provided by the present invention is a composition comprising an antibody to an above-described polypeptide or an anti-antibody to an above-described polypeptide. Any embodiment of the present invention including the present inventive polypeptide, nucleic acid, antibody, and anti-antibody, can be incorporated into a composition comprising a carrier. The carrier can serve any function. For example, the carrier can increase the solubility of the present inventive polypeptide, nucleic acid or antibody in aqueous solutions. Additionally, the carrier can protect the present inventive polypeptide, nucleic acid or antibody from environmental insults, such as dehydration, oxidation, and photolysis. Moreover, the carrier can serve as an adjuvant, or as a timed-release control means in a biological system.

Antibodies can be generated in accordance with methods known in the art. See, for example, Benjamin, In Immunology: a short course, Wiley-Liss, NY, 1996, pp. 436-437; Kuby, In Immunology, 3rd. ed. , Freeman, NY, 1997, pp. 455-456; Greenspan et al., FASEB J. 7: 437-443 (1993); and Poskitt, Vaccine 9: 792-796 (1991). Anti- antibodies (i.e., anti-idiotypic antibodies) also can be generated in accordance with methods known in the art (see, for example, Benjamin, In Immunology: a short course, Wiley-Liss, NY, 1996, pp. 436-437; Kuby, In Immunology, 3rd. ed., Freeman, NY, 1997, pp. 455-456; Greenspan et al., FASEB J. , 7, 437-443, 1993; Poskitt, Vaccine, 9, 792-796, 1991; and Madiyalakan et al., Hybridonor 14: 199-203 (1995) ("Anti-idiotype induction therapy" ) ) . Such antibodies can be obtained and employed either in solution-phase or coupled to a desired solid- phase matrix. Having in hand such antibodies, one skilled in the art will further appreciate that such antibodies, using well-established procedures (e.g., such as described by Hariow and Lane (1988, supra) , are useful in the detection, quantification, or purification of gpl20 or HIV, particularly HIV-1, conjugates of each and host cells transformed to produce a gpl20 receptor or a derivative thereof. Such antibodies are also useful in a method of prevention or treatment of a viral infection and in a method of inducing an immune response to HIV as provided herein.

In view of the above, an above-described polypeptide can be administered to an animal. The animal generates anti-polypeptide antibodies. Among the anti-polypeptide antibodies generated or induced in the animal are antibodies that have an internal image of gpl20. In accordance with well-known methods, polyclonal or monoclonal antibodies can be obtained, isolated and selected. Selection of an anti-polypeptide antibody that has an internal image of gpl20 can be based upon competition between the anti-polypeptide antibody and gpl20 for binding to an above-described polypeptide, or upon the ability of the anti-polypeptide antibody to bind to a free polypeptide as opposed to a polypeptide bound to gpl20. Such an anti-antibody can be administered to an animal to prevent or treat an HIV infection in accordance with methods provided herein.

Although nonhuman anti-idiotypic antibodies, such as an anti-polypeptide antibody that has an internal image of gpl20 and, therefore, is anti-idiotypic to gpl20, are useful for prophylaxis in humans, their favorable properties might, in certain instances, can be further enhanced and/or their adverse properties further diminished, through "humanization" strategies, such as those recently reviewed by Vaughan, Nature Biotech.. 16, 535-539, 1998.

Prior to administration to an animal, such as a mammal, in particular a human, an above-described polypeptide, nucleic acid, antibody or anti-antibody can be formulated into various compositions by combination with appropriate carriers, in particular, pharmaceutically acceptable carriers or diluents, and can be formulated to be appropriate for either human or veterinary applications . The present invention also provides a method of making an antibody. The method comprises administering an immunogenic amount of an above-described polypeptide or nucleic acid to an animal, such as a mammal, in particular a human. Determining the quantity of a polypeptide or nucleic acid that is immunogenic will depend in part on the degree of similarity to a protein or other molecule of the inoculated animal, the route of administration of the polypeptide or nucleic acid, and the size of the polypeptide administered or encoded by the administered nucleic acid. If necessary, the polypeptide or nucleic acid can be mixed with or ligated to a substance (or an adjuvant) that enhances its i munogenicity. Such calculations and procedures are within the skill of the ordinary artisan. Additionally, the present inventive method preferably can be used to induce an immune response against HIV, particularly HIV-l, in a mammal, particularly a human.

In view of the above, the present invention further provides a method of prophylactically or therapeutically treating an HIV infection in a mammal, particularly a human, in need thereof. The method comprises administering to the mammal an HIV replication-inhibiting effective amount of an above-described polypeptide, nucleic -acid, or an anti-antibody to an above-described polypeptide or a nucleic acid encoding such a polypeptide .

The present invention also provides a method of prophylactically or therapeutically treating HIV infection in a mammal. The method comprises administering to the mammal an effective amount of an above-described polypeptide or nucleic acid. Prior to administration to an animal, such as a mammal, in particular a human, an above-described polypeptide or nucleic acid can be formulated into various compositions by combination with appropriate carriers, in particular, pharmaceutically acceptable carriers or diluents, and can be formulated to be appropriate for either human or veterinary applications .

Thus, a composition for use in the method of the present invention can comprise one or more of the polypeptides, nucleic acids, antibodies or anti- antibodies described herein, preferably in combination with a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well-known to those skilled in the art, as are suitable methods of administration. The choice of carrier will be determined, in part, by whether a polypeptide or a nucleic acid is to be administered, as well as by the particular method used to administer the composition. Optionally, the carrier can be selected to increase the solubility of the composition or mixture, e.g., a liposome or polysaccharide. One skilled in the art will also appreciate that various routes of administering a composition are available, and, although more than one route can be used for administration, a particular xoute can provide a more immediate and more effective reaction than another route. Accordingly, there are a wide variety of suitable formulations of compositions that can be used in the present inventive methods.

A composition in accordance with the present invention, alone or in further combination with one or more other active agents, can be made into a formulation suitable for parenteral administration, preferably intraperitoneal administration. Such a formulation can include aqueous and nonaqueous, isotonic sterile injection solutions, which can contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and nonaqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The formulations can be presented in unit dose or multi-dose sealed containers, such as ampules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, water, for injections, immediately prior to use. Extemporaneously injectable solutions and suspensions can be prepared from sterile powders, granules, and tablets,- as described herein.

A formulation suitable for oral administration can consist of liquid solutions, such as an effective amount of the compound dissolved in diluents, such as water, saline, or fruit juice; capsules, sachets or tablets, each containing a predetermined amount of the active ingredient, as solid or granules; solutions or suspensions in an aqueous liquid; and oil-in-water emulsions or water-in-oil emulsions. Tablet forms can include one or more of lactose, mannitol, corn starch, potato starch, microcrystalline cellulose, acacia, gelatin, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, stearic acid, and other excipients, colorants, diluents, buffering agents, moistening agents, preservatives, flavoring agents, and pharmacologically compatible carriers.

Similarly, a formulation suitable for oral administration can include lozenge forms, which can comprise the active ingredient in a flavor, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier; as well as creams, emulsions, gels, and the like containing, in addition to the active ingredient, such carriers as are known in the art. An aerosol formulation suitable for administration via inhalation also can be made. The aerosol formulation can be placed into a pressurized acceptable propellant, such as dichlorodifluoromethane, propane, nitrogen, and the like.

A formulation suitable for topical application can be in the form of creams, ointments, or lotions.

A formulation for rectal administration can be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate. A formulation suitable for vaginal administration can be presented as a pessary, tampon, cream, gel, paste, foam, or spray formula containing, in addition to the active ingredient, such carriers as are known in the art to be appropriate .

Important general considerations for design of delivery systems and compositions, and for routes of administration, for polypeptide drugs also apply (Eppstein, CRC Crit. Rev. Therapeutic Drug Carrier Systems 5, 99-139, 1988; Siddiqui et al . , CRC Crit. Rev. Therapeutic Druσ Carrier Systems 3, 195-208, 1987); Banga et al., Int. J. Pharmaceutics 48, 15-50, 1988; Sanders, Eur. J. Druσ Metab. Pharmacokinetics 15, 95-102, 1990; Verhoef, Eur. J. Drug Metab. Pharmacokinetics 15, 83-93, 1990) . The appropriate delivery system for a given polypeptide will depend upon its particular nature, the particular clinical application, and the site of drug action. As with any protein drug, oral delivery will likely present special problems, due primarily to instability in the gastrointestinal tract and poor absorption and bioavailability of intact, bioactive drug therefrom. Therefore, especially in the case of oral delivery, but also possibly in conjunction with other routes of delivery, it will be necessary to use an absorption-enhancing agent in combination with a given polypeptide. A wide variety of absorption-enhancing agents have been investigated and/or applied in combination with protein drugs for oral delivery and for delivery by other routes (Verhoef, 1990, supra: van Hoogdalem, Pharmac. Ther. 44, 407-43, 1989; Davis, O . Pharm. Pharmacol. 44 (Suppl. 1), 186-90, 1992). Most commonly, typical enhancers fall into the general categories of (a) chelators, such as EDTA, salicylates, and N-acyl derivatives of collagen, (b) surfactants, such as lauryl sulfate and polyoxyethylene-9-lauryl ether, (c) bile salts, such as glycholate and taurocholate, and derivatives, such as taurodihydrofusidate, (d) fatty acids, such as oleic acid and capric acid, and their derivatives, such as acylcarnitines, monoglycerides, and diglyσerides, (e) non-surfactants, such as unsaturated cyclic ureas, (f) saponins, (g) cyclodextrins, and (h) phospholipids.

Other approaches to enhancing oral delivery of protein drugs can include the aforementioned chemical modifications to enhance stability to gastrointestinal enzymes and/or increased lipophilicity. Alternatively, the protein drug can be administered in combination with other drugs or substances that directly inhibit proteases and/or other potential sources of enzymatic degradation of proteins. Yet another alternative approach to prevent or delay gastrointestinal absorption of protein drugs is to incorporate them into a delivery system that is designed to protect the protein from contact with the proteolytic enzymes in the intestinal lumen and to release the intact protein only upon reaching an area favorable for its absorption. A more specific example of this strategy is the use of biodegradable microcapsules or microspheres, both to protect vulnerable drugs from degradation, as well as to effect a prolonged release of active drug (Deasy, iβ Microencapsulation and Related Processes. Swarbrick, ed., Marcell Dekker, Inc.: New York, 1984, pp. 1-60, 88-89, 208-11) . Microcapsules also can provide a useful way to effect a prolonged delivery of a protein drug after injection (Maulding, J Controlled Release 6, 167-76, 1987).

The dose administered to an animal, such as a mammal, particularly a human, in the context of the present invention should be sufficient to effect a therapeutic or prophylactic response in the individual over a reasonable time frame. The dose will be determined by the particular polypeptide, nucleic acid, antibody, or anti-antibody administered, the severity of any existing disease state, as well as the body weight and age of the individual. The size of the dose also will be determined by the existence of any adverse side effects that may accompany the use of the particular polypeptide, nucleic acid, antibody or anti-antibody employed. It is always desirable, whenever possible, to keep adverse side effects to a minimum.

The dosage can be in unit dosage form, such as a tablet or capsule. The term "unit dosage form" as used herein refers to physically discrete units suitable as unitary dosages for human and animal subjects, each unit containing a predetermined quantity of a vector, alone or in combination with other active agents, calculated in an amount sufficient to produce the desired effect in association with a pharmaceutically acceptable diluent, carrier, or vehicle. The specifications for the unit dosage forms of the present invention depend on the particular embodiment employed and the effect to be achieved, as well as the pharmacodynamics associated with each polypeptide, nucleic acid or anti-antibody in the host. The dose administered should be an "HIV infection inhibiting amount" of an above-described polypeptide or nucleic acid or an "immune response-inducing effective amount" of an above-described polypeptide, an above- described nucleic acid, or an antibody as appropriate. Another composition provided by the present invention is a composition comprising a solid support matrix to which is attached an above-described polypeptide, or an anti-antibody to an above-described polypeptide. The solid matrix can comprise other functional reagents including, for example, polyethylene glycol, dextran, albumin and the like, whose intended e fector functions may include one or more of the following: to improve stability of the conjugate; to increase the half-life of the conjugate; to increase resistance of the conjugate to proteolysis; to decrease the immunogenicity of the conjugate; to provide a means to attach or immobilize a functional polypeptide or anti- antibody onto a solid support matrix (e.g., see, for example, Harris, in PolvfEthylene Glycol⁾ Chemistry; Biotechn-i al and Biomedical Applications, Harris, ed. , Plenum Press: New York (1992), pp. 1-14). Conjugates furthermore may comprise a polypeptide or anti-antibody coupled to an effector molecule, each of which, optionally, may have different functions (e.g., such as a toxin molecule (or an immunological reagent) and a polyethylene glycol (or dextran or albumin) molecule) . Diverse applications and uses of functional proteins and polypeptides, attached to or immobilized on a solid support matrix, are exemplified more specifically for poly(ethylene glycol) conjugated proteins or peptides in a review by Holmberg et al. (In Polv(.Ethylene Glvcol) Chemistry: Biotechnical and Biomedical Applications. Harris, ed., Plenum Press: New York, 1992, pp. 303-324). In addition, the present invention provides a method of removing HIV from a bodily fluid of an animal. The method comprises extracorporeally contacting the bodily fluid of the animal with a solid-support matrix to which is attached an above-described polypeptide or an anti- antibody to an above-described polypeptide. Alternatively, the bodily fluid can be contacted with the polypeptide or anti-antibody in solution and then .the solution can be contacted with a solid support matrix to which is attached a means to remove the polypeptide or anti-antibody to which is bound HIV gpl20 from the bodily fluid. Methods of attaching an herein-described polypeptide, or an anti-antibody to a solid support matrix are known in the art. "Attached" is used herein to refer to attachment to (or coupling to) and immobilization in or on a solid support matrix. See, for example, Harris, in Poly (Ethylene Glvcol) Chemistr^y; Biotechnical and Biomedical Applications, Harris, ed. , Plenum Press: New York (1992), pp. 1-14) and international patent application WO 91/02714 (Saxinger) . Diverse applications and uses of functional polypeptides attached to or immobilized on a solid support matrix are exemplified more specifically for poly(ethylene glycol) conjugated proteins or peptides in a review by Holmberg et al. (In Polv (Ethylene Glycol⁾ Chemistry; Biotechnical and Biomedical Applications, Harris, ed., Plenum Press: New York, 1992, pp. 303-324). The present invention also provides a method of making an antibody that binds to gpl20 of HIV under physiological conditions. The method comprises labeling an embodiment of the present inventive compound to obtain a labeled compound. Labeling compounds are within the skill of the ordinary artisan. For example, the present inventive compound can be labeled with radioactive atom, such as ^ιasI in the same or a similar manner as was performed in the examples provided below. Alternatively, an enzyme, such as horseradish peroxidase, can be attached to or incorporated into the present inventive compound. Then by exposing a chromogenic or photogenic compound to the compound, a signal indicative of the presence and quantity of the compound present can be generated. In another alternative, a polyhistidinyl moiety can be attached to, or incorporated into, the present inventive moiety so that the present inventive compound will react with high affinity to transition metal ions such as nickel, copper, or zinc ions; this reaction can be used as the basis to quantify the amount of the present inventive compound present at a particular location. In yet another alternative, the present inventive compound can be used as antigen to a standard antibody that specifically recognizes an antigenic epitope of the present inventive compound. As is well- known, the standard antibody can itself be labeled or used in conjunction with an additional antibody that is labeled with an enzyme, radioisotope, or other suitable means. The skilled artisan will recognize that there is a plethora of other suitable means and methods to label the present inventive compound. This present inventive method of making an antibody that binds to a gpl20 envelope protein of HIV further comprises providing a library of synthetic peptides. The library consists of a multiplicity of synthetically- produced polypeptides that are homologous, and preferably essentially identical (i.e., having the same primary amino acid residue sequence, ignoring blocking groups, phosphorylation of serinyl, threoninyl, and tyrosinyl residues, hydroxylation of prolinyl residues, and the like) or identical, to a continuous region of an HIV gp_l2₀ envelope protein. The .polypeptides of the library can be any suitable length. While larger regions allow faster scanning and tend to preserve non-linear epitopes; shorter length polypeptides allow more sensitive screening of the primary sequence of the gpl20 protein. However, polypeptides that are too short can lose essential secondary structure or cleave reactive sites into one or more pieces. Preferably, a mixture of short and long polypeptides are incorporated into the library, however, the library can consist of polypeptides of a single length (measured in amino acid residues) . For the sake of convenience the library can be split into multiple parts, and screened by parts. Typically, the polypeptides of the library will be between about 6 and about 45 amino acid residues in length.

Typically, the library will comprise a series of polypeptides each having an identical sequence to that of gpl20 but having an amino-terminus a particular number of amino acids downstream of the amino-terminus of the prior polypeptide (see, examples section below) . The distance, measured in amino acid residues, is referred to as the offset. Preferably, libraries that are characterized by the existence of an offset, the offset is not greater than the product of length of the longest polypeptide measured in amino acid residues and 1.5, preferably 1.0, and more preferably 0.5. The library can be alternatively characterized by the existence of an offset not greater than 30, preferably 15, and more preferably 4.

Each polypeptide of the library is substantially isolated from every other polypeptide of said library and is located in a known position. For example, each polypeptide can be bound to a solid support and that is in a vessel or that can be placed in a vessel. The vessel preferably enables each polypeptide to be covered in a liquid that does not contact any other oligonucleotide of the library. By way of example, each polypeptide can be bound to a bead that is placed in a vessel (or tube) or can be bound to the well of a multi- well assay plate. Alternatively, an array of polypeptides can be fashioned, for example on a microchip device (as is presently used in some DNA sequencing devices and methods) , and the entire array can be bathed in a single solution.

Each polypeptide is then individually contacted with the labeled compound such that a portion of the labeled compound can bind with the polypeptide of the library. In this way, a bound population of each labeled compound of the present invention and an unbound population of the labeled compound is generated. The phrase individually contacted means that each polypeptide has the opportunity to bind with the labeled compound and the quantity of labeled compound bound by each can be determined.

The method then comprises removing substantially all of the unbound labeled compound from the position occupied by each polypeptide. That is, the solution comprising the labeled compound is separated from the polypeptides of the library and the bound population of the labeled compound. This can be done by any suitable method, e..g., by aspiration and one or more washing steps comprising adding a quantity of liquid sufficient to cover all the surfaces that were contacted by the labeled compound and aspirating away substantially all of the wash liquid.

The amount of labeled compound that remains co-localized with each polypeptide of the library is then measured to determine the quantity of labeled compound bound by each polypeptide. The amount of the present inventive compound bound by each polypeptide can be directly evaluated to identify a portion of the HIV gpl20 envelope protein that binds to an (HIV) -receptor selected from the group consisting of CCR5, CXCR4, STRL33, and CD4. This information is then used to identify and provide an immunizing compound. The immunizing compound comprises a polypeptide comprising an amino acid sequence that is homologous to, or preferably is essentially identical to, or identical to, the portion of the HIV-1 gpl20 envelope protein that binds with CD4, CCR5, CXCR4, and/or STRL33. The immunizing protein can be provided by processing gpl20, e.g., proteolytically digesting gpl20 that has been isolated from a preparation of HIV-1. Preferably, however, the immunizing compound is prepared synthetically, or by genetic engineering, or by a combination of genetic engineering and synthetic methods. The immunizing compound can comprise a pharmaceutically acceptable substituent, can be encoded by a nucleic acid that can be expressed in a cell, can be mixed with a carrier, and is an inventive aspect of the present invention.

An immunogenic quantity of the immunizing compound is then inserted into an animal (e.g., a human, or a rodent, a canine, a feline, or a ruminant) in a manner consistent with the discussion of a method of raising an antibody to the present inventive compounds that are homologous to portions of CCR5, CXCR4, STRL33, and CD4, above. The insertion of the immunizing compound causes the inoculated animal to produce an antibody that binds with said portion of the HIV gpl20 envelope protein.

Thus the present invention also provides an antibody that binds to an HIV gpl20 envelope protein, as well as an antigen binding protein comprising one or more complementarity determining regions of the antibody (e.g., a Fab, a Fab₂,, an Fv, a single-chain antibody, a diabody, and humanized variants of all of the above, all of which are within the skill in the art) .

The antibody or variant thereof is preferably useful in detecting or diagnosing the presence of HIV gpl20 envelope protein, and thus HIV, in an animal. The antibody is also preferably prevents or attenuates infection of an animal exposed to HIV, to whom an effective quantity of the antibody or a variant thereof, has been administered or produced in response to inoculation with the immunizing compound. The antibody preferably also is useful in treating or preventing (i.e., inhibiting) HIV infection in an animal to whom a suitable dose has been administered or in which a suitable quantity of antibody has been produced. The antibody is also useful in the study of HIV infection of mammalian cells, the host range specificities of HIV infection, and preferably, the mechanism by which antibodies neutralize infectious viruses.

EXAMPLES

The following examples further illustrate the present invention but, of course, should not be construed as limiting the scope of the claimed invention in any way. Synthetic peptide arrays were constructed in 96-well microtiter plates in accordance with the method set forth in WO 91/02714 (Saxinger) , and used to test the binding of HIV-l_u envelope gpl20 that had been labeled with radioactive iodine (radiolabeling by standard methods) . After incubating the radiolabeled gpl20 in a well with each. synthetic peptide, a washing step was performed to remove unbound label, and the relative level of radioactivity remaining in each well of the plate was evaluated to determine the relative affinity of each peptide for the gpl20. The synthesis of the peptides and the quantity of binding between the synthetic peptides and the gpl20 were found to be suitably reproducible, precise, and sensitive. Initial screening of the entire primary sequence of the chemokine and CD4 receptor molecules was taken 18 amino acid residues at a time. The authenticity of the binding signals generated by this technique has been repeatedly demonstrated by showing that antibodies to CCR5 and CXCR4 are able to inhibit the binding of radiolabeled gpl20 to the polypeptides derived from CCR5 and CXCR4 that show a high affinity for binding with gpl20. Additionally, the accuracy of the binding assay used hereinbelow is demonstrated by Example 7.

Example 1 This example identifies segments of the CCR5 co-receptor that bind with gpl20.

The first column in the table below indicates the number of the amino acid in the wild-type CCR5 receptor. The second column explicitly identifies the peptide sequence. The third column indicates the radioactive counts recorded in twenty minutes (i.e., the cpm x 20) after the background or non-specific counts had been subtracted. The fourth column contains an X in each row for which the listed polypeptide bound with high affinity to gρl20. The fifth and final column contains an X in each row wherein the listed sequence binds with substantial affinity but is weak in comparison to other samples, particularly adjacent samples.

SEQ SEG PEPTIDE Counts Peak non- Peak per 20' Activity activity

Average- background e pty (control) 7 f

1- -18 MDYQVSSPIYDINYYTSE 73J X

5- -22 VSSPIYDINYYTSEPCQK 382 X

9- -26 lYDINYYTSEPCQKINVK 228 X

13 -30 NYYTSEPCQKINVKQIAA e

17-34 SEPCQKINVKQIAARLLP -44 1-38 QKINVKQIAARLLPPLYS 20 5 -42 VKQIAARLLPPLYSLVFI 18 9-46 AARLLPPLYSLVFIFGFV 33 3 -50 LPPLYSLVFIFGFVGNML 705 X 7-54 YSLVFIFGFVGNMLVILI 347 X 1-58 FIFGFVGNMLVILILINC 343 X 5-62 FVGNMLVILILINCKRLK 62 9-66 MLVILILINCKRLKSMTD 84 3 -70 LILINCKRLKSMTDIYLL 2 7-74 NCKRLKSMTDIYLLNLAI 25 1-78 LKSMTDIYLLNLAISDLF 210 5-82 TDI YLLNLAI SDLFFLLT 38 9-86 LLNLAI SDLFFLLTVPFW 144 3 -90 AISDLFFLLTVPFWAHYA 41 7-94 LFFLLTVPFWAHYAAAQW 173 1-98 LTVPFWAHYAAAQWDFGN 306 5- FWAHYAAAQWDFGNTMCQ 212 9- YAAAQWDFGNTMCQLLTG 494 X 3 - QWDFGNTMCQLLTGLYFI 1019 X 7- GNTMCQLLTGLYFIGFFS 941 X 01- CQLLTGLYFIGFFSGIFF 489 X 05- TGLYFIGFFSGIFFI ILL 80 09- FIGFFSGIFFIILLTIDR 76 13- FSGIFFIILLTIDRYLAV 83 17- FFI ILLTIDRYLAWHAV 77 21- LLTIDRYLAWHAVFALK 31 25- DRYLAWHAVFALKARTV 62 29- AWHAVFALKARTVTFGV 34 33 - AVFALKARTVTFGWTSV 63 137- LKARTVTFGWTSVITWV 74

141- TVTFGWTSVITWWAVF -25

145- GWTSVITWWAVFASLP 69

149- SVITWWAVFASLPGIIF 46

153- WWAVFASLPGIIFTRSQ 871

157- VFASLPGIIFTRSQKEGL 541

161- LPGIIFTRSQKEGLHYTC 118

165- IFTRSQKEGLHYTCSSHF

169- SQKEGLHYTCSSHFPYSQ 304 x

173- GLHYTCSSHFPYSQYQFW 301 x

177- TCSSHFPYSQYQFWKNFQ 367 X

181- HFPYSQYQFWKNFQTLKI 1008 X

185- SQYQFWKNFQTLKIVILG 15721

189- FWKNFQTLKIVILGLVLP 40

193- FQTLKIVILGLVLPLLVM 45

197- KIVILGLVLPLLVMVICY 65

201- LGLVLPLLVMVICYSGIL 180

205- LPLLVMVICYSGILKTLL 68

209- VMVICYSGILKTLLRCRN -8

213- CYSGILKTLLRCRNEKKR 70

217- ILKTLLRCRNEKKRHRAV 19

221- LLROiNEKKRHRAVRLIF 1021

225- RNEKKRHRAVRLIFTIMI 23

229- KRHRAVRLIFTIMIVYFL 36

233- AVRLIFTIMIVYFLFWAP 62

237- IFTIMIVYFLFWAPYNIV 121

241- MIVYFLFWAPYNIVΓ.T.T.N 214

245- FLFWAPYNIVLLLNTFQE 616 X

249- APYNIVLLLNTFQEFFGL 1962 X

253- IVTiTiT.NTFQEFFGLNNCS 2134 X

257- LNTFQEFFGLNNCSSSNR 293

261- QEFFGLNNCSSSNRLDQA 63

265- GLNNCSSSNRLDQAMQVT -31

269- CSSSNRLDQAMQVTETLG 90

273- NRLDQAMQVTETLGMTHC 10

277- QAMQVTETLGMTHCCINP 81

281- VTETLGMTHCCINPIIYA 15

285- LGMTHCCINPIIYAFVGE 282 X

289- HCCINPIIYAFVGEKFRN 200 X

293- NPIIYAFVGEKFRNYLLV 162 X

297- YAFVGEKFRNYLLVFFQK 596

301- GEKFRNYLLVFFQKHIAK 69 305- RNYLLVFFQKHIAKRFCK 65

309- LVFFQKHIAKRFCKCCSI 761

313- QKHIAKRFCKCCSIFQQE 23

317- AKRFCKCCSIFQQEAPER 64

321- CKCCSIFQQEAPERASSV 53

325- SIFQQEAPERASSVYTRS 100

329- QEAPERASSVYTRSTGEQ 84

333- ERASSVYTRSTGEQEISV 84

337- SVYTRSTGEQEISVGL 47

These data indicate that, in addition to polypeptide sequences derived from positions 1-18 of the CCR5 receptor, the polypeptide sequences LPPLYSLVFIFGFVGNML, QWDFGNTMCQLLTGLYFIGFFS, SQYQFWKNFQTLKIVILG,

APYNIVLLLNTFQEFFGLNNCS, and YAFVGEKFRNYLLVFFQK comprise multiple subsequences, each which is capable of binding to HIV-1 envelope gpl20.

Example 2

This example identifies segments of the CXCR4 co-receptor that bind with gpl20.

The first column in the table below indicates the number of the amino acid in the wild-type CXCR4 receptor. The second column explicitly identifies the peptide sequence. The third and fourth columns indicate the radioactive counts recorded in twenty minutes (i.e., the cpm x 20) after the background or non-specific counts had been subtracted. The fifth column contains an X in each row for which the listed polypeptide bound with high affinity to gpl20. The sixth and final column contains an X in each row wherein the listed sequence binds with substantial affinity but is weak in comparison to other samples, particularly adjacent samples. SEQ SEG PEPTIDE M jor Minor

Activity Activity

Peak Peak empty (control)

1— 18 MEGISIYTSDNYTEEMGS

5--22 SIYTSDNYTEEMGSGDYD

9--26 SDNYTEEMGSGDYDSMKE

13-30 TEEMGSGDYDSMKEPCFR

17-34 GSGDYDSMKEPCFREENA

21-38 YDSMKEPCFREENANFNK

25-42 KEPCFREENANFNKIFLP

29-46 FREENANFNKIFLPTIYS

33-50 NANFNKIFLPTIYSIIFL

37-54 NKIFLPTIYSIIFLTGIV

41-58 LPTIYSIIFLTGIVGNGL

45-62 YSIIFLTGIVGNGLVILV

49-66 FLTGIVGNGLVILVMGYQ

53-70 IVGNGLVILVMGYQKKLR

57-74 GLVILVMGYQKKLRSMTD

61-78 LVMGYQKKLRSMTDKYRL

65-82 YQKKLRSMTDKYRLHLSV

69-86 L SMTDKYRLHLSVADLL

73-90 TDKYRLHLSVADLLFVIT

77-94 RLHLSVADLLFVITLPFW

81-98 SVADLLFVITLPFWAVDA

85-102 LLFVITLPFWAVDAVANW

89-106 ITLPFWAVDAVANWYFGN

93-110 FWAVDAVANWYFGNFLCK

97-114 DAVANWYFGNFLCKAVHV

101-118 NWYFGNFLCKAVHVIYTV

105-122 GNFLCKAVHVIYTVNLYS

109-126 CKAVHVIYTVNLYSSVLI

113-130 HVIYTVNLYSSVLILAFI

117-134 TVNLYSSVLILAFISLDR

121-138 YSSVLILAFISLDRYLAI

125-142 LILAFISLDRYLAIVHAT

129-146 FISLDRYLAIVHATNSQR

13 -150 DRYLAIVHATNSQRPRKL

137-154 AIVHATNSQRPRKLLAEK

141-158 ATNSQRPRKLLAEKWYV

145-162 QRPRKLLAEKWYVGVWI

149-166 KLLAEKWYVGVWIPALL 153-170 EKWYVGVWIPALLLTIP 157-174 YVGVWIPALLLTIPDFIF 161-178 WIPALLLTIPDFIFANVS 165-182 LLLTIPDFIFANVSEADD 169-186 IPDFIFANVSEADDRYIC 173-190 IFANVSEADDRYICDRFY 177-194 VSEADDRYICDRFYPNDL 181-198 DDRYICDRFYPNDLWWV 185-202 ICDRFYPNDLWWVFQFQ 189-206 FYPNDLWVWFQFQHIMV 193-210 DLWVWFQFQHIMVGLIL 197-214 WFQFQHIMVGLILPGIV 201-218 FQHIMVGLILPGIVILSC 205-222 MVGLILPGIVILSCYCII 209-226 ILPGIVILSCYCIIISKL 213-230 IVILSCYCIIISKLSHSK 217-234 SCYCIIISKLSHSKGHQK 221-238 IIISKLSHSKGHQKRKAL 225-242 KLSHSKGHQKRKALKTTV 229-246 SKGHQKRKALKTTVILIL 233-250 QKRKALKTTVILILAFFA 237-254 ALKTTVILILAFFACWLP 241-258 TVILILAFFACWLPYYIG 245-262 ILAFFACWLPYYIGISID 249-266 FACWLPYYIGISIDSFIL 253-270 LPYYIGISIDSFI LEII 257-274 IGISIDSFILLEIIKQGC 261-278 IDSFILLEIIKQGCEFEN 265-282 ILLEIIKQGCEFENTVHK 269-286 IIKQGCEFENTVHKWISI 273-290 GCEFENTVHKWISITEAL 277-294 ENTVHKWISITEALAFFH 281_.-298 HKWISITEALAFFHCCLN 285-302 SITEALAFFHCCLNPILY 289-306 ALAFFHCCLNPILYAFLG 293-310 FHCCLNPILYAFLGAKFK 297-314 LNPILYAFLGAKFKTSAQ 301-318 LYAFLGAKFKTSAQHALT 305-322 LGAKFKTSAQHALTSVSR 309-326 FKTSAQHALTSVSRGSSL 313-330 AQHALTSVSRGSSLKILS

317-334 LTSVSRGSSLKILSKGKR

321-338 SRGSSLKILSKGKRGGHS

325-3 2 SLKILSKGKRGGHSSVST

329-346 LSKGKRGGHSSVSTESES

333-350 KRGGHSSVSTESESSSFH

337-352 HSSVSTESESSSFHSS

These data indicate that, in addition to polypeptide sequences derived from positions 1-18 of the CXCR4 receptor, the polypeptide sequences LLLTIPDFIFANVSEADD (165-182) , WFQFQHIMVGLILPGIV (197-214) , and IDSFILLEIIKQGCEFEN (261-278) comprise multiple subsequences, which is capable of binding to HIV-1 envelope gpl20.

Example 3

This example identifies segments of the STRL33 co-receptor that bind with gpl20.

The first column in the table below indicates the number of the amino acid in the wild-type STRL33 receptor. The second column explicitly identifies the peptide sequence. The third and fourth columns indicate the radioactive counts recorded in twenty minutes ⁽i.e., the cpm x 20) after the background or non-specific counts had been subtracted. The fifth column contains an X in each row for which the listed polypeptide bound with high affinity to gpl20. The sixth and final column contains an X in each row wherein the listed sequence binds with substantial affinity but is weak in comparison to other samples, particularly adjacent samples. Major Minor

SEQ SEG PEPTIDE Activity Activity

Peak Peak

empty (control)

1--18 MAEHDYHEDYGFSSFNDS X

5--22 DYHEDYGFSSFNDSSQEE X

9--26 DYGFSSFNDSSQEEHQAF

13-30 SSFNDSSQEEHQAFLQFS X

17-34 DSSQEEHQAFLQFSKVFL X

21-38 EEHQAFLQFSKVFLPCMY X

25-42 AFLQFSKVFLPCMYLWF X

29-46 FSKVFLPCMYLWFVCGL

33-50 FLPCMYLWFVCGLVGNS

37-54 MYLWFVCGLVGNSLVLV

41-58 VFVCGLVGNSLVLVISIF

45-62 GLVGNSLVLVISIFYHKL

49-66 NSLVLVISIFYHKLQSLT

53-70 LVISIFYHKLQSLTDVFL X

57-74 IFYHKLQSLTDVFLVNLP X

61-78 KLQSLTDVFLVNLPLADL

65-82 LTDVFLVNLPLADLVFVC

69-86 FLVNIiPLADLVFVCTLPF

73-90 LPLADLVFVCTLPFWAYA X

77-94 DLVFVCTLPFWAYAGIHE X

81-98 VCTLPFWAYAGIHEWVFG X

85-102 PFWAYAGIHEWVFGQVMC

89-106 YAGIHEWVFGQVMCKSLL X

93-110 HEWVFGQVMCKSLLGIYT X

97-114 FGQVMCKSLLGIYTINFY X

101-118 MCKSLLGIYTINFYTSML

105-122 LLGIYTINFYTSMLILTC

109-126 YTINFYTSMLILTCITVD

113-130 FYTSMLILTCITVDRFIV

117-134 MLILTCITVDRFIVWKA

121-138 TCITVDRFIVWKATKAY

125-142 VDRFIWVKATKAYNQQA

129-146 IVWKATKAYNQQAKRMT

133-150 KATKAYNQQAKRMTWGKV

137-15 AYNQQAKRMTWGKVTSLL

141-158 QAKRMTWGKVTSLLIWVI

145-162 MTWGKVTSLLIWVISLLV

149 -166 KVTSLLIWVISLLVSLPQ

153 - 170 LLIWVISLLVSLPQIIYG

157-174 VI SLLVSLPQI I YGNVFN

161-178 LVSLPQIIYGNVFNLDKL

165 -182 PQIIYGNVFNLDKLICGY

169 -186 YGNVFNLDKLICGYHDEA

173 -190 FNLDKLICGYHDEAISTV X

177- 194 KLICGYHDEAISTWLAT X

181-198 GYHDEAIST LATQMTL X

185 -202 EAISTWLATQMTLGFFL

189 -206 TWLATQMTLGFFLPLLT X

193 -210 ATQMTLGFFLPLLTMIVC X

197-214 TLGFFLPLLTMIVCYSVI

201-218 FLPLLTMIVCYSVIIKTL X

205-222 LTMIVCYSVIIKTLLHAG

209 -226 VCYSVIIKTLLHAGGFQK X

213 -230 VI IKTLLHAGGFQKHRSL

217-234 TLLHAGGFQKHRSLKIIF

221-238 AGGFQKHRSLKIIFLVMA

225 -242 QKHRSLKIIFLVMAVFLL

229-246 SLKIIFLVMAVFLLTQMP

233 -250 IFLVMAVFLLTQMPFNLM

237 -254 MAVFLLTQMPFNLMKFIR X

241-258 LLTQMPFNLMKFIRSTHW X

245-262 MPFNLMKFI STHWEYYA X

249-266 LMKFIRSTHWEYYAMTSF

253 -270 IRSTHWEYYAMTSFHYTI X

257-274 HWEYYAMTSFHYTIMVTE

261-278 YAMTSFHYTIMVTEAIAY X

265 -282 SFHYTIMVTEAIAYLRAC

269-286 TIMVTEAIAYLRACLNPV

273 -290 TEAI AYLRACLNPVLYAF

277-294 AYLRACLNPVLYAFVSLK X

281-298 ACLNPVLYAFVSLKFRKN

285 -302 PVLYAFVSLKFRKNFWKL X

289-306 AFVSLKFRKNFWKLVKDI

293 -310 LKFRKNFWKLVKDIGCLP

297-314 KNFWKLVKDIGCLPYLGV

301-318 KLVKDIGCLPYLGVSHQW

305 -322 DIGCLPYLGVSHQWKSSE

309-326 LPYLGVSHQWKSSEDNSK

313 -330 GVSHQWKSSEDNSKTFSA

317-334 QWKSSEDNSKTFSASHNV 321-338 SEDNSKTFSASHNVEATS 325-342 SKTFSASHNVEATSMFQL

These data indicate that, in addition to polypeptide sequences derived from positions 9-26 of the STRL33 receptor, the polypeptide sequences LVISIFYHKLQSLTDVFL (53-70) , PFWAYAGIHEWVFGQVMC (85-102) , EAISTWLATQMTLGFFL (185-202) , LTMIVCYSVIIKTLLHAG (205-222) , MAVFLLTQMPFNLMKFIRSTHW (237-258), HWEYYAMTSFHYTIMVTE (257-274) , ACLNPVLYAFVSLKFRKN (281-298) and SKTFSASHNVEATSMFQL (325-342) comprise multiple subsequences, which is capable of binding to HIV-1 envelope gpl20.

Example 4

This example identifies segments of the human CD4 protein that bind with gpl20.

The second column in the in the table below identifies the amino acid residue sequence of the polypeptide employed in the assay. The first column identifies the sequence coordinates of human CD4 that have an identical amino acid sequence. The third column indicates the number of radioactive decays (i.e., counts) that were counted, which is indicative of the affinity of the synthetic polypeptide for the gpl20 protein. In the table below, polypeptides retaining more than 4,000 counts identify fragments that have a substantial capability to bind with gpl20. Polypeptides retaining more than 6,000 counts have more substantial binding affinity. Polypeptides retaining at least about 10,000 counts have a substantial and strong capacity to bind to gpl20. Of course, fragments corresponding to amino acid coordinates 101-121 and 106-126 have a substantial, strong, and dominant capacity to bind to gpl20.

Bl ( 1) 1-21 MNRGVPFRHLLLVLQLALLPA 3587

CI ( 2) 6-26 PFRHLLLVLQLALLPAATQGK 4355

Dl ( 3) 11-31 LLVLQLALLPAATQGKKWLG 1735

El ( 4) 16-36 LALLPAATQGKKWLGKKGDT 759

FI ( 5) 21-41 AATQGKKWLGKKGDTVELTC 1562

Gl ( 6) 26-46 KKWLGKKGDTVELTCTASQK 1910

HI ( 7) 31-51 GKKGDTVELTCTASQKKSIQF 1831

A2 ( 8) 36-56 TVELTCTASQKKSIQFHWKNS 1732

B2 ( 9) 41-61 CTASQKKSIQFHWKNSNQIKI 1717

C2 ( 10) 4,6-66 KKSIQFHWKNSNQIKILGNQG 2182

D2 < 11) ^■ 51-71 FHWKNSNQIKILGNQGSFLTK 1835

Ξ2 ( 12) 56-76 SNQIKILGNQGSFLTKGPSKL 1487

F2 ( 13) 61-81 ILGNQGSFLTKGPSKLNDRAD 1467

G2 < 14) 66-86 GSFLTKGPSKLNDRADSRRSL 1844

H2 < 15) 71-91- KGPSKLNDRADSRRSLWDQGN 1912

A3 < 16) 76-96 LNDRADSRRSLWDQGNFPLII 1753

B3 < 17) 81-101 DSRRSLWDQGNFPLIIKNLKI 2224

C3 < 18) 86-106 LWDQGNFPLIIKNLKIEDSDT 3264

D3 ( 19) 91-111 NFPLIIKNLKIEDSDTYICEV 11646

E3 ( 20) 96-116 IKNLKIEDSDTYICEVEDQKE 8439

F3 | 21) 101-121 IEDSDTYICEVEDQKEEVQLL 6803

G3 ( 22) 106-126 TYICEVEDQKEEVQLLVFGLT 44965

H3 |^r23) 111-131 VEDQKEEVQLLVFGLTANSDT 36249

A4 I 24) 116-136 EEVQLLVFGLTANSDTHLLQG 14171

B4 [25) 12Ϊ.-141 LVFGLTANSDTHLLQGQSLTL 3683

C4 [26) 126-146 TANSDTHLLQGQSLTLTLESP 6114

D4 [27) 131-151 THLLQGQSLTLTLESPPGSSP 2552

Ξ4 [28) 136-156 GQSLTLTLESPPGSSPSVQCR 1538

F4 [29) 141-161 LTLESPPGSSPSVQCRSPRGK 1476

G4 [30) 146-166 PPGSSPSVQCRSPRGKNIQGG 1496

H4 [3D 151-171 PSVQCRSPRGKNIQGGKTLSV 1400

A5 [32) 156-176 RSPRGKNIQGGKTLSVSQLEL 2066

B5 (33) 161-181 KNIQGGKTLSVSQLELQDSGT 3078

C5 (34) 166-186 GKTLSVSQLELQDSGTWTCTV 2618

D5 (35) 171-191 VSQLELQDSGTWTCTVLQNQK 3879

E5 (36) 176-196 LQDSGTWTCTVLQNQKKVEFK 2456

F5 (37) 181-201 TWTCTVLQNQKKVEFKIDIW 4030

G5 (38) 186-206 VLQNQKKVEFKIDIWLAFQK 9737

H5 (39) 191-211 KKVEFKIDIWLAFQKASSIV 6313

A6 (40) 196-216 KIDIWLAFQKASSIVYKKEG 3681 201-221 VLAFQKASSIVYKKEGEQVEF 3566

206-226 KASSIVYKKEGEQVEFSFPLA 14347

211-231 VYKKEGEQVEFSFPLAFTVEK 14740

216-236 GEQVEFSFPLAFTVEKLTGSG 18549

221-241 FSFPLAFTVEKLTGSGELWWQ 9673

226-246 AFTVEKLTGSGELWWQAERAS 3992

231-251 KLTGSGELWWQAERASSSKSW 1878

236-256 GELWWQAERASSSKSWITFDL 2730

241-261 QAERASSSKSWITFDLKNKEV 2588

246-266 SSSKSWITFDLKNKEVSVKRV 1761

251-271 WITFDLKNKEVSVKRVTQDPK 2126

256-276 LKNKEVSVKRVTQDPKLQMGK 2288

261-281 VSVKRVTQDPKLQMGKKLPLH 1848

266-286 VTQDPKLQMGKKLPLHLTLPQ 2075

271-291 KLQMGKKLPLHLTLPQALPQY 1949

276-296 KKLPLHLTLPQALPQYAGSGN 1922

281-301 HLTLPQALPQYAGSGNLTLAL 2394

286-306 QALPQYAGSGNLTLALEAKTG 2364

291-311 YAGSGNLTLALEAKTGKLHQE 1830

296-316 NLTLALEAKTGKLHQEVNLW 1676

301-321 LEAKTGKLHQEVNLWMRATQ 1729

306-326 GKLHQEVNLWMRATQLQKNL 1776

311-331 EVNLWMRATQLQKNLTCEVW 2183

316-336 VMRATQLQKNLTCEVWGPTSP 2144

32Ϊ-341 QLQKNLTCEVWGPTSPKLMLS 1856

326-346 LTCEVWGPTSPKLMLSLKLEN 2412

331-351 WGPTSPKLMLSLKLENKEAKV 2414

336-356 PKLMLSLKLENKEAKVSKREK 1656

341-361 SLKLENKEAKVSKREKAVWVL 1663

346-366 NKEAKVSKREKAVWVLNPEAG 1735

351-371 VSKREKAVWVLNPEAGMWQCL 2034

356-376 KAVWVLNPEAGMWQCLLSDSG 3133

361-381 LNPEAGMWQCLLSDSGQVLLE 6316

366-386 GMWQCLLSDSGQVLLESNIKV 4185

371-391 LLSDSGQVLLESNIKVLPTWS 2375

376-396 GQVLLESNIKVLPTWSTPVQP 2089

381-401 ESNIKVLPTWSTPVQPMALIV 1992

386-406 VLPTWSTPVQPMALIVLGGVA 2197

391-411 STPVQPMALIVLGGVAGLLLF 2527

396-416 PMALIVLGGVAGLLLFIGLGI 3067

401-421 VLGGVAGLLLFIGLGIFFCVR 3738

406-426 AGLLLFIGLGIFFCVRCRHRR 2099

411-431 FIGLGIFFCVRCRHRRRQAER 1900

416-436 IFFCVRCRHRRRQAERMSQIK 2085

421-441 RCRHRRRQAERMSQIKRLLSE 2075

426-446 RRQAERMSQIKRLLSEKKTCQ 1607 Hll(87) 431-451 RMSQIKRLLSEKKTCQCPHRF 2020

A12(88) 436-456 KRLLSEKKTCQCPHRFQKTCS 1674

B12(89) 441-458 EKKTCQCPHRFQKTCSPI 2006

Al ( 0) empty (control) 2075

Example 5

This example shows the binding of "'i-HIV-l^ gpl20 to the amino termini of CCR5, CXCR4, and STRL33 as a function of the dependence on position and length- Synthetic peptide arrays of nonapeptides, dodecapeptides, pentadecapeptides and octadecapeptides derived from CCR5 (panel A) , CXCR4 (panel B) and STRL33 (panel C) amino terminal domains were prepared and utilized to test the binding of ^XMI-HIV-1_« envelope gpl20. Ordinal sequence position numbers are given in accordance with the sequence data provided by the Genbank database for CCR5 (accession No. gl457946, gi|l457946), CXCR4 (accession No. g539677, gi|400654, sp|P30991) and STRL33 (accession No. g2209288, gi|2209288). The counts shown are the counts detected in each well minus the background counts (i.e., counts observed in the assay when no polypeptide was bound to the well of the 96-well assay plate) .

Panel A Peptide Sequence Scanning Binding Results For Window Length

Windows CCR5 (counts bound - background (no peptide))

(In each sequence row 9-,

Initial 12-, 15-, 18-mers share the

Sequence same initial starting point)

^'# xxxxxxxxx 9 9 xxxxxxxxxxxx 12 12 xxxxxxxxxxxxxx 15 15 xxxxxxxxxxxxxxxxxx 18 18

1 MDYQVSSPIYDINYYTSE 543 2682 4976 5880

2 DYQVSSPIYDINYYTSEP 1552 3089 5401 6363

3 YQVSSPIYDINYYTSEPC 2533 5305 5415 6119

4 QVSSPIYDINYYTSEPCQ 490 1959 4594 5645

5 VSSPIYDINYYTSEPCQK 509 1629 3280 3521

6 SSPIYDINYYTSEPCQKI 671 1739 3498 3285

7 SPIYDINYYTSEPCQKIN 1503 3463 4575 3234

8 PIYDINYYTSEPCQKINV 1186 2285 2682 2036

9 lYDINYYTSEPCQKINVK 1359 2702 2516 1261

10 YDINYYTSEPCQKINVKQ 4379 5245 3052 1913

11 DINYYTSEPCQKINVKQI 1396 1361 1144 712

12 INYYTSEPCQKINVKQIA 1384 1190 707 684

13 NYYTSEPCQKINVKQIAA 1548 977 760 595

14 YYTSEPCQKINVKQIAAR 1029 1052 847 638

15 YTSEPCQKINVKQIA 567 507 459

16 TSEPCQKINVKQIAA 440 427 509

17 SEPCQKINVKQIAAR 434 430 426

18 EPCQKINVKQIA 397 432

19 PCQKINVKQIAA 386 385

20 CQKENVKQIAAR 435 581

21 QKINVKQIA 453

22 KINVKQIAA 487

23 INVKQIAAR 474

Panel B Peptide Sequence Scanning Binding Results For Window

Windows Length

CXCR4

(In each sequence row 9-, 12-, 15-, 18- (counts bound -1 background) mers share the same initial starting point)

Initial

Sequence ά xxxxxxxxx 9 9 xxxxxxxxxxxx 12 12 xxxxxxxxxxxxxxx 15 15 xxxxxxxxxxxxxxxxxx 18 18

1 MEGISIYTSD YTEEMGS 591 334 3275 2079

2 EGISIYTSDNYTEEMGSG a 886 7255 1548

3 GISIYTSDNYTEEMGSGD 454 2644 3274 1217

4 ISIYTSDNYTΕEMGSGDY 466 3973 2202 861

5 SIYTSDNYTEEMGSGDYD a 288 168 239

6 IYTSDNYTEEMGSGDYDS 332 335 195 173

7 YTSDNYTEEMGSGDYDSM 181 161 201 103 a

8 TSDNYTEEMGSGDYDSMK 54 119 38

9 SDNYTEEMGSGDYDSMKE 151 149 124 161

10 DNYIΕEMGSGDYDSM EP 67 121 57 102

11 NYTEEMGSGDYDSMKEPC a 100 30 134

12 YTEEMGSGDYDSMKEPCF 68 213 70 103

13 TEEMGSGDYDSMKEPCFR 146 67 23 47

14 EEMGSGDYDSMKEPCFRE a 61 121 130

15 EMGSGDYDSMKEPCFREE 64 36 69 64

16 MGSGDYDSMKEPCFREEN 57 68 64 129

17 GSGDYDSMKEPCFREENA a 155 172 155

18 SGDYDSMKEPCFREENAN 100 118 186 89

19 GDYDSMKEPCFREENANF 53 167 198 134 a

20 DYDSMKEPCFREENANFN 167 146 75

21 YDSMKEPCFREENANFNK 171 144 80 89

22 DSMKEPCFREENANFNKI 85 144 146 40 a

23 SMKEPCFREENANFN 119 55

24 MKEPCFREENANFNK 188 133 74

25 KEPCFREENANFNKI 165 105 93 a

26 EPCFREENANFN 69

27 PCFREENANFNK 104 108

28 CFREENANFNKI 103 66

29 REENANFNK 58 a ot done Panel C Peptide Sequence Scanning Binding Results For Window Length

Windows

ST L33 (counts bound - background)

(In each sequence row 9-, 12- Initial 15-, 18-mers share the same Sequence # initial starting point.)

xxxxxxxxx 9 9 xxxxxxxxxxxx 12 12 xxxxxxxxxxxxxxx 15 15 xxxxxxxxxxxxxxxxxxl8 18

1 MAEHDYHEDYGFSSFNDS 160 625 1239 1386

2 AEHDYHEDVGFSSFNDSS 354 697 1095 1014

3 EHDYHEDYGFSSFNDSSQ 509 937 2235 1219

4 HDYHEDYGFSSFNDSSQE 708 1427 1772 1500

5 DYHEDYGFSSFNDSSQEE 851 1554 1240 1191

6 YHEDYGFSSFNDSSQEEH 728 1950 1357 985

7 HEDYGFSSFNDSSQEEHQ 729 1077 947 537

8 EDYGFSSFNDSSQEEHQA 953 817 1152 548

9 DYGFSSFNDSSQEEHQAF 701 573 595 440

10- YGFSSFNDSSQEEHQAFL 345 745 645 1138

11 GFSSFNDSSQEEHQAFLQ 171 480 270 1639

12 FSSFNDSSQEEHQAFLQF 249 403 361 3608

13 SSFNDSSQEEHQAFLQFS 243 277 902 6038

14 SFNDSSQEEHQAFLQFSK 304 303 969 4537

15 FNDSSQEEHQAFLQFSKV 246 470 4089 4678

16 NDSSQEEHQAFLQFS 180 497 6160

17 DSSQEEHQAFLQFSK 147 882 4588

18 SSQEEHQAFLQFSKV 287 4455 4732

19 SQEEHQAFLQFS 647 7512

20 QEEHQAFLQFSK 1109 5672

21 EEHQAFLQFSKV 6060 5598

22 EHQAFLQFS 7505

23 HQAFLQFSK 2761

24 QAFLQFSKV 2600

Example 6

_This example shows '"i-HIV-l^ gpl20 binding to _N-terminal peptide variants of CCR5 , CXCR4 and STRL33 Octadecapeptide alanine replacement variants of maximum gpl20 binding activity peaks were synthesized and tested for ^12SI-HIV-1_IAI gpl20 binding. Each binding value presented is the average of two separate synthesis and binding experiments. Relative percentage of Control = {[(mean counts/Control counts)] x 100%} ± average deviation. Background counts (no peptide, see Example 7⁾ were subtracted from all values. Data for CCR5 are presented in Panel A; data for CXCR4 are presented in Panel B; and data for STRL33 are presented in Panel C.

Panel A. ^I-HTV-I_LAI gpl20 binding to N-terminal peptide variants of CCR5

CCR5 variantpeptides (1-18) Relative % ofControl^a

Control MDY_QVSSPIYDINYYTSE 100

MIA ADY_QVSSPIYDINYYTSE 167 ± 4

D2A MAY_QVSSPIYDINYYTSE 125 ± 8

Y3A MDA_QVSSPIYDINYYTSE 51 ± 2

Q4A MDYAVSSPIYDINYΎTSE 104 ± 7

V5A MDY_QASSPIYDINYYTSE 82 ± 3

S6A MDY_QVASPIYDINYΎTSE 124 ± 3

S7A MDY_QVSAPIYDINYYTSE 56 ± 2

P8A MDY_QVSSAIYDINYYTSE 157 ± 2

I9A MDY_QVSSPAYDINYYTSE 24 ± 7

Y10A MDY_QVSSPIADINYYTSE 19 ± 6

D11A MDY_QVSSPIYAINYYTSE 63 ± 22

I12A MDY_QVSSPIYDANYYTSE 14 ± 1

N13A MDY_QVSSPIYDIAYYTSE 253 ± 19

Y14A MDY_QVSSPIYDINAYTSE 15 ± 0.3

Y15A MDY_QVSSPIYDINYATSE 21 ± 5

T16A MDY_QVSSPIYDINYYASE 78 ± 34

S17A MDY_QVSSPIYDINYYTAE 64 ± 6

E18A MDY_QVSSPIYDINYYTSA 4 ± 2 Panel B ^I-HTV-I_LA! gpl20 bindingtoN-terminalpeptide variants of CXCR4

CXCR4 variantpeptides (1-18) Relative % ofControl⁸

Control MEGISIYTSDNYTEEMGS 100

MIA AEGISIYTSDNYTEEMGS 118 ± 18

E2A MAGISIYTSDNYTEEMGS 36 ± 0.3

G3A MEAISIYTSDNYTEEMGS 101 -fc 3

I4A MEGASIYTSDNYTEEMGS 6 ± 03

S5A MEGIAIYTSDNYTEEMGS 133 ^■k 5

I6A MEGISAYTSDNYTEEMGS - 1

Y7A MEGISIATSDNYTEEMGS 7 ± 0.4

T8A MEGISIYASDNYTEEMGS 97 ± 10

S9A MEGISIYTADNYTEEMGS 70 -fc 4

D10A MEGISIYTSANYTEEMGS 71 -b 8

NilA MEGISIYTSDAYTEEMGS 38 ± 0.4

Y12A MEGISIYTSDNATEEMGS 28 ± 2

T13A MEGISIYTSDNYAEEMGS 70 ± 6

E14A MEGISIYTSDNYTAEMGS 72 ± 1

E15A MEGISIYTSDNYTEAMGS 56 ± 7

M16A MEGISIYTSDNYTEEAGS 88 ± 4

G17A MEGISIYTSDNYTEEMAS 68 ± 8

S18A MEGISIYTSDNYTEEMGA 79 ± 1

¹ The percent binding for the wild-type peptide was- defined as 100%.

Panel C ^I-HTV-I_LA! gpl20 binding to N-terminal peptide variants of

STRL33

STRL33 variant peptides (21-38) Relative % of Control⁸

Control EEHQAFLQFSKVFLPCMY 100

E21A AEHQAFLQFS VFLPCMY 81 ± 2

E22A EAHQAFLQFSKVFLPCMY 70 ± 1

H23A EEAQAFLQFSKVFLPCMY 99 ± 1

Q24A EEHAAFLQFSKVFLPCMY 72 ± 1

A25A EEHQAFLQFSKVFLPCMY 101 ± 1

F26A EEHQAALQFSKVFLPCMY 32 ± 0.1

L27A EEHQAFAQFSKVFLPCMY 37 ± 2

Q28A EEHQAFLAFSKVFLPCMY 44 ± 0.4

F29A EEHQAFLQASKVFLPCMY 20 ± 1

S30A EEHQAFLQFAKVFLPCMY 92 ± 2

K31A EEHQAFLQFSAVFLPCMY 162 ± 2

V32A EEHQAFLQFSKAFLPCMY 51 ± 3

F33A EEHQAFLQFSKVALPCMY 45 ± 2

L34A EEHQAFLQFSKVFAPCMY 76 ± 1

P35A EEHQAFLQFSKVFLACMY 82 ± 3

C36A EEHQAFLQFSKVFLPAMY 53 ± 5

M37A EEHQAFLQFSKVFLPCAY 112 ± 4

Y38A EEHQAFLQFSKVFLPCMA 83 ± 2 ^a The percent binding for the wild-type peptide was i defined as 100%.

Example 7 This example demonstrates that the binding of HIV-1 gpl20 envelope protein to the polypeptides of the present invention and to the chemokine receptors from which the present inventive polypeptides were originally derived or inspired is conserved across the various species of HIV-1. This example also demonstrates that a step subsequent to initial binding of gpl20 to CCR5, CXCR4, STRL33, and CD4 is the most likely source of the phenomenon of host-range selectivity. Additionally, this example demonstrates that the underlying method is accurate in that receptor variants that are predicted to have an altered affinity for binding with gpl20, do in fact have a statistically similar alteration in affinity where comparable changes in the receptors have been identified in other work and the affinity for binding of gpl20/effect on infectivity has been measured. This example examines the effect of particular mutations of CCR5 that were studied in the work underlying the present invention and that were also studied by other artisans in the field.

The following table identifies a mutation in the first column. The first letter designates the wild-type amino acid present at the position indicated by the number, and the letter A which terminates all entries in the first column indicates that the amino acid residue present in that position in the mutant polypeptide is alaninyl. For example, the first data row (i.e., the second row of the table) contains the entry Y3A in the first column, which indicates that the tyrosine residue at position 3 of the wild-type CCR5 is substituted by an alanine residue. The second column provides the percentage of binding exhibited by a mutant polypeptide compared to a wild-type polypeptide, when the methods used to elucidate the present invention are used in conjunction with radiolabeled HIV-l^ gpl20 envelope protein. The third through seventh columns provide similar data that have been extracted from the work of others in the field using a strain of HIV-1 virus indicated at the top of each column. For example, row 2 of the following table indicates that when the mutation Y3A is effected in the human CCR5 chemokine receptor, then the resulting CCR5 polypeptide has 51.4% of the ability to bind HIV-l^ gpl20 envelope protein in comparison to an equivalent wild-type peptide. Similarly, HIV-1^ binds to the mutant polypeptide with 79% of the affinity of a non-mutated CCR5 chemokine receptor.

Statistical analysis of these data indicates that the similarity between the binding affinity of each mutant peptide for gpl20 elucidated in this study is not more than about 25% likely to be causally unrelated to the effects observed for YU2, and not more than about 4% likely to be causally unrelated to the effects observed for each of the other viruses listed in the table above.

Additionally, the affinity measurements generated by the underlying technique has been demonstrated to be accurate by (repetitively) showing that antibodies that specifically bind to radiolabeled gpl20 are capable of preventing the binding of gpl20 to polypeptides that have shown high affinity for binding with gpl20 in the experiments upon which the present invention is predicated. Thus, this example shows that the binding with chemokine receptors HIV-1 can be inhibited by the present inventive polypeptides, irrespective of the strain of HIV-1 from which the gpl20 protein is obtained. Example 8

This example provides a characterization of the critical amino acids in the amino-terminal segments of CCR5, CXCR4, and STRL33 that are essential for the ability of these polypeptides to bind with gpl20.

In this example, the effect on binding that occurs to due successive replacement of each amino acid with alanine is indicated, wherein a (+) signifies a decrease in binding affinity and a (>) signifies an enhancement in

10 binding affinity. As is clear from inspection, the sequences are shown with that amino-terminus at top and the carboxyl-terminus at bottom.

Example 9

This example employs the same technique as Example 4 and provides information similar to that available from Example 4.

The data below compares the ability of synthetic fragments of CD4 to bind to labeled gpl20. 9-mer, 12-mer, 15-mer, 18-mer, and 21-mers were selected based on the data from Examples 4. The relative binding affinities of each group of polypeptides can be determined by inspection of the number of counts of radiolabeled gpl20 that were retained by each N-mer. Data supporting these conclusions are provided by Examples 10 and 11.

112 EDQKEEVQLLVFGLT 3633

113 DQKEEVQLLVFGLTA 3905

114 QKEEVQLLVFGLTAN 3770

115 KEEVQLLVFGLTANS 3485

116 ΞEVQLLVFGLTANSD 6423

117 EVQLLVFGLTANSDT 2689

130 DTHLLQGQSLTLTLE 1622

131 THLLQGQSLTLTLES 1874

132 HLLQGQSLTLTLESP 1277

213 KKEGEQVEFS FPLAF 1921

214 KEGEQVEFSFPLAFT 3253

215 EGEQVEFSFPLAFTV 3270

216 GEQVEFS FPLAFTVE 4656

217 EQVEFSFPLAFTVEK 4135

218 QVEFSFPLAFTVEKL 2047!

ACTIVE21-MERS

90 GNFPLIIKNLKIEDS 5248 DTYICE

91 NFPLIIKNLKIEDSD 7803

TYICEV 92 FPLIIKNLKIEDSDT 13919

YICEVE 93 PLIIKNLKIEDSDTY 20145

ICEVED 94 LIIKNLKIEDSDTYI 17108

CEVEDQ 95 IIKNLKIEDSDTYIC 11892

EVEDQK 96 IKNLKIEDSDTYICE 15073

VEDQKE 97 KNLKIEDSDTYICEV 8789

EDQKEE

89ILKIEDSDTYICEVED 5519

QKEEVQ 100IKIEDSDTYICEVEDQ 6325

KEEVQL 101IIEDSDTYICEVEDQK 12064

EEVQLL 102IEDSDTYICEVEDQKE 4933

EVQLLV 103IDSDTYICEVEDQKEE 30277

VQLLVF 104|SDTYICEVEDQKEEV 30319

QLLVFG 105IDTYICEVEDQKEEVQ 25424

LLVFGL 106ITYICEVEDQKEEVQL 20191

LVFGLT 107|γiCEVEDQKEEVQLL 22884

VFGLTA 1081ICEVEDQKEEVQLLV 7276

FGLTAN 109ICEVEDQKEEVQLLVF 3517

GLTANS

123 FGLTANSDTHLLQGQ 11529

SLTLTL

124 GLTANSDTHLLQGQS 14065

LTLTLE

125 LTANSDTHLLQGQSL 17113

TLTLES

126 TANSDTHLLQGQSLT 23595

LTLESP

Example 10

This example provides data which enables those skilled in the art to arrive at the conclusions indicated in Examples 9 and 12. In this example, the counts of radiolabeled gp-120 retained by each peptide indicated in the left hand column are given in the right hand.column. The first panel (panel A) provides data for 21-mers of CD4.

Panel A PEPTIDE COUNTS

L D_QGNFPLIIKNLKIEDSDT 731 WDQGNFPLIIKNLKIEDSDTY 889 DQGNFPLIIKNLKIEDSDTYI 1138 QGNFPLIIKNLKIEDSDTYIC 2242

GNFPLIIKNLKIEDSDTYICE 5248

NFPLIIKNLKIEDSDTYICEV 7803

FPLIIKNLKIEDSDTYICEVE 13919

PLIIKNLKIEDSDTYICEVED 20145

LIIKNLKIEDSDTYICEVEDQ 17108

IIKNLKIEDSDTYICEVEDQK 11892

IKNLKIEDSDTYICEVEDQKE 15073

KNLKIEDSDTYICEVEDQKEE 8789

NLKIEDSDTYICEVEDQKEEV 2016

LKIEDSDTYICEVEDQKEEVQ 5519

KIEDSDTYICEVEDQKEEVQL 6325

IEDSDTYICEVEDQKEEVQLL 12064

EDSDTYICEVEDQKEEVQLLV 4933

DSDTYICEVED_QKEEVQLLVF 30277

SDTYICEVEDQKEEVQLLVFG 30319

DTYICEVED_QKEEVQLLVFGL 25424

TYICEVED_QKEEVQLLVFGLT 20191

YICEVED_QKEEVQLLVFGLTA 22884

ICEVED_QKEEVQLLVFGLTAN 7276

CEVED_QKEEVQLLVFGLTANS 3517

EVED_QKEEV_QLLVFGLTANSD 1687

VED_QKEEV_QLLVFGLTANSDT 646

ED_QKEEV_QLLVFGLTANSDTH 562

D_QKEEV_QLLVFGLTANSDTHL 599

_QKEEV_QLLVFGLTANSDTHLL 573

KEEV_QLLVFGLTANSDTHLLQ 682

EEV_QLLVFGLTANSDTHLLQG 690

EV_QLLVFGLTANSDTHLLQGQ 589

V_QLLVFGLTANSDTHLLQGQS 1099

QLLVFGLTANSDTHLLQGQSL 2057

LLVFGLTANSDTHLLQGQSLT 860

LVFGLTANSDTHLLQGQSLTL 4677

VFGLTANSDTHLLQGQSLTLT 2762

FGLTANSDTHLLQGQSLTLTL 11529

GLTANSDTHLL_QGQSLTLTLE 14065

LTANSDTHLL_QGQSLTLTLES 17113

TANSDTHLL_QGQSLTLTLESP 23595

Empty (Control) 515

TWTCTVL_QNQKKVEFKIDIW 1430

WTCTVL_QNQKKVEFKIDIWL 1616

TCTVLQN_QKKVEFKIDIWLA 1092

CTVL_QN_QKKVEFKIDIWLAF 2909

TVL_QN_QKKVEFKIDIWLAFQ 3273

VLQN_QKKVEFKIDIWLAFQK 1323 L_QNQKKVEFKIDIWLAFQKA 1256

QN_QKKVEFKIDIWLAFQKAS 1808

NQKKVEFKIDIWLAFQKASS 1507

QKKVEFKIDIWLAFQKASSI 759

KKVEFKIDIWLAFQKASSIV 782

KVEFKIDIWLAFQKASSIVY 635

VEFKIDIWLAFQKASSIVYK 725

EFKIDIWLAFQKASSIVYKK 649

FKIDIWLAF_QKASSIVYKKE 593

KIDIWLAFQKASSIVYKKEG 1394

IDIWLAFQKASSIVYKKEGE 962

DIWLAFQKASSIVYKKEGEQ 788

IWLAF_QKASSIVYKKEGEQV 646

WLAFQKASSIVYKKEGEQVE 772

VLAF_QKASSIVYKKEGEQVEF 1793

LAF_QKASSIVYKKEGEQVEFS 1410

AF_QKASSIVYKKEGEQVEFSF 3775

F_QKASSIVYKKEGEQVEFSFP 9382

_QKASSIVYKKEGEQVEFSFPL 24959

KASSIVYKKEGEQVEFSFPLA 30873

ASSIVYKKEGEQVEFSFPLAF 25146

SSIVYKKEGEQVEFSFPLAFT 28068

SIVYKKEGEQVEFSFPLAFTV 8165

IVYKKEGE_QVEFSFPLAFTVE 15620

VYKKEGE_QVEFSFPLAFTVEK 2429

YKKEGE_QVEFSFPLAFTVEKL 735

KKEGEQVEFSFPLAFTVEKLT 1847

JKEGE_QVEFSFPLAFTVEKLTG 972

EGE_QVEFSFPLAFTVEKLTGS 739

GE_QVEFSFPLAFTVEKLTGSG 652

E_QVEFSFPLAFTVEKLTGSGE 765

QVEFSFPLAFTVEKLTGSGEL 741

VEFSFPLAFTVEKLTGSGELW 633

EFSFPLAFTVEKLTGSGELWW 681

FSFPLAFTVEKLTGSGELWWQ 4163

SFPLAFTVEKLTGSGEL QA 2284

FPLAFTVEKLTGSGEL WQAE 6276

PLAFTVEKLTGSGEL WQAER 2647

LAFTVEKLTGSGELW QAERA 3577

AFTVEKLTGSGEL QAERAS 1739

Empty (control) 617 These second and third panels (panels B and C) provide data for 18-mers of a small region of CD4.

PanelB

PEPTIDE COUNTS

LWDQGNFPLIIKNLK 502

WDQGNFPLIIKNLKI 534

DQGNFPLIIKNLKIE 635

QGNFPLIIKNLKIED 509

GNFPLIIKNLKIEDS 624

NFPLIIKNLKIEDSD 654

FPLIIKNLKIEDSDT 539

PLIIKNLKIEDSDTY 661

LIIKNLKIEDSDTYI 542

IIKNLKIEDSDTYIC 664

IKNLKIEDSDTYICE 568

KNLKIEDSDTYICEV 562

NLKIEDSDTYICEVE 1160

LKIEDSDTYICEVED 846

KIEDSDTYICEVEDQ 1088

IEDSDTYICEVEDQK 1143

EDSDTYICEVEDQKE 815

DSDTYICEVEDQKEE 973

SDTYICEVEDQKEEV 993

DTYICEVEDQKEEVQ 1071

TYICEVEDQKEEVQL 956

YICEVEDQKEEVQLL 1064

ICEVEDQKEEVQLLV 1084

CEVEDQKEEVQLLVF 1729

EVEDQKEEVQLLVFG 2805

VEDQKEEVQLLVFGL 3816

EDQKEEVQLLVFGLT 3633

DQKEEVQLLVFGLTA 3905

QKEEVQLLVFGLTAN 3770

'KEEVQLLVFGLTANS 3485

EEVQLLVFGLTANSD 6423

EVQLLVFGLTANSDT 2689

VQLLVFGLTANSDTH 1006

QLLVFGLTANSDTHL 865

LLVFGLTANSDTHLL 599

LVFGLTANSDTHLLQ 609

VFGLTANSDTHLLQG 532

FGLTANSDTHLLQGQ 625 GLTANSDTHLLQGQS 532

LTANSDTHLLQGQSL 634

TANSDTHLLQGQSLT 513

ANSDTHLLQGQSLTL 542

NSDTHLLQGQSLTLT 631

SDTHLLQGQSLTLTL 747

DTHLLQGQSLTLTLE 1622

THLLQGQSLTLTLES 1874

HLLQGQSLTLTLESP 1277 LWDQGNFPLIIKNLKIED 582

WDQGNFPLIIKNLKIEDS 626

DQGNFPLIIKNLKIEDSD 598

QGNFPLIIKNLKIEDSDT 564

GNFPLIIKNLKIEDSDTY 557

NFPLIIKNLKIEDSDTYI 627

FPLIIKNLKIEDSDTYIC 509

PLIIKNLKIEDSDTYICE 624

LIIKNLKIEDSDTYICEV 634

IIKNLKIEDSDTYICEVE 751

IKNLKIEDSDTYICEVED 699

KNLKIEDSDTYICEVEDQ 708

NLKIEDSDTYICEVEDQK 863

LKIEDSDTYICEVEDQKE 872

KIEDSDTYICEVEDQKEE 858

IEDSDTYICEVEDQKEEV 1230 EDSDTYICEVEDQKEEVQ 788

DSDTYICEVEDQKEEVQL 961

SDTYICEVEDQKEEVQLL 870

DTYICEVEDQKEEVQLLV 1648

TYICEVEDQKEEVQLLVF 3794

YICEVEDQKEEVQLLVFG 4611

ICEVEDQKEEVQLLVFGL 3898

CEVEDQKEEVQLLVFGLT 3797

EVEDQKEEVQLLVFGLTA 3647

VEDQKEEVQLLVFGLTAN 3913

EDQKEEVQLLVFGLTANS 3416

DQKEEVQLLVFGLTANSD 3317

QKEEVQLLVFGLTANSDT 3671

KEEVQLLVFGLTANSDTH 1271

EEVQLLVFGLTANSDTHL 783

EVQLLVFGLTANSDTHLL 667

VQLLVFGLTANSDTHLLQ 673

QLLVFGLTANSDTHLLQG 574

LLVFGLTANSDTHLLQGQ 568

TVFGLTANSDTHLLQGQS 564 VFGLTANSDTHLLQGQSL 531

FGLTANSDTHLLQGQSLT 591

GLTANSDTHLLQGQSLTL 572

LTANSDTHLLQGQSLTLT 528

TA SDTHLLQGQSLTLTL 891

ANSDTHLLQGQSLTLTLE 1540

NSDTHLLQGQSLTLTLES 1726

SDTHLLQGQSLTLTLESP 1260

Empty (control) 575

Panel C

PEPTIDE COUNTS

TCTVLQNQKKVEFK 566

TCTVLQNQKKVEFKI 510

CTVLQNQKKVEFKID 608

TVLQNQKKVEFKIDI 587

VLQNQKKVEFKIDIV 605

LQNQKKVEFKIDIW 644

QNQKKVEFKIDIWL 636

NQKKVEFKIDIWLA 860

QKKVEFKIDIWLAF 1333

KKVEFKIDIWLAFQ 951

KVEFKIDIWLAFQK 1051

VEFKIDIWLAFQKA 1005

EFKIDIWLAFQKAS 1188

FKIDIWLAFQKASS 1001

KIDIWLAFQKASSI 956

IDIWLAFQKASSIV 865

DIWLAFQKASSIVY 776

IWLAFQKASSIVYK 783

WLAFQKASSIVYKK 577

VLAFQKASSIVYKKE 634

LAFQKASSIVYKKEG 593

AFQKASSIVYKKEGE 544

FQKASSΓVYKKEGEQ 637

QKASSIVYKKEGEQV 519

KASSIVYKKEGEQVE 563

ASSIVYKKEGEQVEF 589

SSIVYKKEGEQVEFS 558

SIVYKKEGEQVEFSF 651

IVYKKEGEQVEFSFP 615

VYKKEGEQVEFSFPL 714 YKKEGEQVEFSFPLA 687

KKEGEQVEFSFPLAF 1921

KEGEQVEFSFPLAFT 3253

EGEQVEFSFPLAFTV 3270

GEQVEFSFPLAFTVE 4656

EQVEFSFPLAFTVEK 4135

QVEFSFPLAFTVEKL 2047

VEFSFPLAFTVEKLT 899

EFSFPLAFTVEKLTG 920

FSFPLAFTVEKLTGS 672

SFPLAFTVEKLTGSG 565

FPLAFTVEKLTGSGE 556

PLAFTVEKLTGSGEL 612

LAFTVEKLTGSGEL 579

AFTVEKLTGSGEL W 586

FTVEKLTGSGELWWQ 625

TVEKLTGSGELWWQA 550

VEKLTGSGELWWQAE 735

EKLTGSGELWWQAER 683 TCTVLQNQKKVEFKIDI 588

TCTVLQNQKKVEFKIDIV 571

CTVLQNQKKVEFKIDIW 553

TVLQNQKKVEFKIDIWL 655

VLQNQKKVEFKIDIWLA 724

LQNQKKVEFKIDIWLAF 938

QNQKKVEFKIDIWLAFQ 917

NQKKVEFKIDIWLAFQK 889

QKKVEFKIDIWLAFQKA 1013

KKVEFKIDIWLAFQKAS 912

KVEFKIDIWLAFQKASS 1011

VEFKIDIWLAFQKASSI 819

EFKIDIWLAFQKASSIV 799

FKIDIWLAFQKASSIVY 843

KIDIWLAFQKASSIVYK 779

IDIWLAFQKASSIVYKK 711

DIWLAFQKASSIVYKKE 660

IWLAFQKASSIVYKKEG 531

WLAFQKASSIVYKKEGE 560

VLAFQKASSIVYKKEGEQ 549

LAFQKASSIVYKKEGEQV 665

AFQKASSIVYKKEGEQVE 514

FQKASSIVYKKEGEQVEF 528

QKASSIVYKKEGEQVEFS 602

KASSIVYKKEGEQVEFSF 536

ASSIVYKKEGEQVEFSFP 701 SSIVYKKEGEQVEFSFPL 756

SIVYKKEGEQVEFSFPLA 771

IVYKKEGEQVEFSFPLAF 5382

VYKKEGEQVEFSFPLAFT 4307

YKKEGEQVEFSFPLAFTV 4839

KKEGEQVEFSFPLAFTVE 4683

KEGE_QVEFSFPLAFTVEK 3117

EGEQVEFSFPLAFTVEKL 2164

GEQVEFSFPLAFTVEKLT 1643

E_QVEFSFPLAFTVEKLTG 798

_QVEFSFPLAFTVEKLTGS 736

VEFSFPLAFTVEKLTGSG 533

EFSFPLAFTVEKLTGSGE 668

FSFPLAFTVEKLTGSGEL 613

SFPLAFTVEKLTGSGELW 656

FPLAFTVEKLTGSGELWW 586

PliAFTVEKLTGSGELWWQ 650

LAFTVEKLTGSGELWWQA 866

AFTVEKLTGSGEL WQAE 788

FTVEKLTGSGELWWQAER 1143

Empty (control) 556

The fourth and fifth panels (Panels D and E) provide data for select 9-mers and 12-mers ofCD4.

PanelD

PEPTIDE COUNTS

DQGNFPLII 662

QGNFPLIIK 508

GNFPLIIKN 600

NFPLIIKNL 561

FPLIIKNLK 601

PLIIKNLKI 697

LIIKNLKIE 515

IIKNLKIED 658

IKNLKIEDS 557

KNLKIEDSD 612

NLKIEDSDT 512

LKIEDSDTY 492

KIEDSDTYI 603

IEDSDTYIC 567

EDSDTYICE 650

DSDTYICEV 712 SDTYICEVE 819

DTYICEVED 1043

TYICEVEDQ 805

YICEVEDQK 728

ICEVEDQKE 596

CEVEDQKEE 555

EVEDQKEEV 587

VEDQKEEVQ 521

EDQKEEVQL 564

DQKEEVQLL 589

QKEEVQLLV 636

KEEVQLLVF 1273

EEVQLLVFG 3170

EVQLLVFGL 2146

VQLLVFGLT 815

QLLVFGLTA 822

LLVFGLTAN 576

LVFGLTANS 522

VFGLTANSD 549

FGLTANSDT 563

GLTANSDTH 481

LTANSDTHL 596

TANSDTHLL 554

ANSDTHLLQ 642

NSDTHLLQG 561

SDTHLLQGQ 526

DTHLLQGQS 578

THLLQGQSL 512

HLLQGQSLT 564

LLQGQSLTL 568

LQGQSLTLT 501

QGQSLTLTL 594

GQSLTLTLE 777

DQGNFPLIIKNL 604

QGNFPLIIKNLK 533

GNFPLIIKNLKI 547

NFPLIIKNLKIE 647

FPLIIKNLKIED 511

PLIIKNLKIEDS 565

LIIKNLKIEDSD 619

IIKNLKIEDSDT 511

IKNLKIEDSDTY 574

KNLKIEDSDTYI 523

NLKIEDSDTYIC 639

LKIEDSDTYICE 635 KIEDSDTYICEV 601

IEDSDTYICEVE 1107

EDSDTYICEVED 956

DSDTYICEVEDQ 937

SDTYICEVEDQK 846

DTYICEVEDQKE 720

TYICEVEDQKEE 818

YICEVEDQKEEV 734

ICEVEDQKEEVQ 585

CEVEDQKEEVQL 561

EVEDQKEEVQLL 508

VEDQKEEVQLLV 657

EDQKEEVQLLVF 1379

DQKEEVQLLVFG 1624

QKEEVQLLVFGL 1785

KEEVQLLVFGLT 1774

EEVQLLVFGLTA 3261

EVQLLVFGLTAN 1838

VQLLVFGLTANS 747

QLLVFGLTANSD 721

LLVFGLTANSDT 533

LVFGLTANSDTH 586

VFGLTANSDTHL 548

FGLTANSDTHLL 571

GLTANPDTHLLQ 574

LTANSDTHLLQG 534

TANSDTHLLQGQ 549

ANSDTHLLQGQS 559

NSDTHLLQGQSL 585

SDTHLLQGQSLT 540

DTHLLQGQSLTL 527

THLLQGQSLTLT 646

HLLQGQSLTLTL 701

LLQGQSLTLTLE 1320

Empty (control) 581

PanelE

PEPTIDE COUNTS

TVLQNQKKV 534

VLQNQKKVE 556

LQNQKKVEF 565

QNQKKVEFK 537

NQKKVEFKI 597 QKKVEFKID 575

KKVEFKIDI 501

KVEFKIDIV 555

VEFKIDIW 548

EFKIDIWL 665

FKIDIWLA 568

KIDIWLAF 665

IDIWLAFQ 691

DIWLAFQK 686

IWLAFQKA 602

WLAFQKAS 600

VLAFQKASS 466

LAFQKASSI 592

AFQKASSIV 595

FQKASSIVY 568

QKASSIVYK 494

KASSIVYKK 498

ASSIVYKKE 600

SSIVYKKEG 515

SIVYKKEGE 566

IVYKKEGEQ 534

VYKKEGEQV 490

YKKEGEQVE 518

KKEGEQVEF 546

KEGEQVEFS 595

EGEQVEFSF 735

GEQVEFSFP 697

EQVEFSFPL 1032

QVEFSFPLA 1205

VEFS^:PLAF 1064

EFSFPLAFT 658

FSFPLAFTV 472

SFPLAFTVE 619

FPLAFTVEK 569

PLAFTVEKL 597

LAFTVEKLT 501

AFTVEKLTG 517

FTVEKLTGS 574

TVEKLTGSG 487

VEKLTGSGE 585

EKLTGSGEL 541

KLTGSGELW 491

LTGSGEL W 550

TGSGELWWQ 507

TVLQNQKKVEFK 563 VLQNQKKVEFKI 503

LQNQKKVEFKID 508

QNQKKVEFKIDI 559

NQKKVEFKIDIV 532

QKKVEFKIDIW 595

KKVEFKIDI L 597

KVEFKIDIWLA 560

VEFKIDIWLAF 681

EFKIDIWLAFQ 659

FKIDIWLAFQK 736

KIDIWLAFQKA 689

IDIWLAFQKAS 630

DIWLAFQKASS 746

IWLAFQKASSI 548

WLAFQKASSIV 567

VLAFQKASSIVY 548

LAFQKASSIVYK 465

AFQKASSIVYKK 597

FQKASSIVYKKE 577

QKASSIVYKKEG 596

KASSIVYKKEGE 559

ASSIVYKKEGEQ 523

SSIVYKKEGEQV 615

SIVYKKEGEQVE 543

IVYKKEGEQVEF 533

VYKKEGEQVEFS 584

YKKEGEQVEFSF 548

KKEGEQVEFSFP 598

KEGEQVEFSFPL 710

EGEQVEFSFPLA 1456

GEQVEFSFPLAF 1729

EQVEFSFPLAFT 1556

QVEFSFPLAFTV 1636

VEFSFPLAFTVE 518

EFSFPLAFTVEK 585

FSFPLAFTVEKL 573

SFPLAFTVEKLT 528

FPLAFTVEKLTG 622

PLAFTVEKLTGS 528

LAFTVEKLTGSG 608

AFTVEKLTGSGE 511

FTVEKLTGSGEL 530

TVEKLTGSGELW 573

VEKLTGSGEL W 477

EKLTGSGEL Q 543 Empty 571 (control)

Panels F and G provide data on sequential alanine replacements for selected CD4 polypeptides.

Panel F

PEPTIDE COUNTS

ZZZZZZDTYICEVED 5844

ZZZZZZATYICEVED 5921

ZZZZZZDAYICEVED 6362

ZZZZZZDTAICEVED 1301

ZZZZZZDTYACEVED 2583

ZZZZZZDTYIAEVED 4483

ZZZZZZDTYICAVED 3154

ZZZZZZDTYICEAED 3432

ZZZZZZDTYICEVAD 3595

ZZZZZZDTYICEVEA 5942

ZZZZZZDTYICEVED 4973

ZZZZZZDTYICEVED 4775

ZZZZZZATYICEVED 4962

ZZZZZZDAYICEVED 4163

ZZZZZZDTAICEVED 1384

ZZZZZZDTYACEVED 3085

ZZZZZZDTYIAEVED 5128

ZZZZZZDTYICAVED 2587

ZZZZZZDTYICEAED 2499

ZZZZZZDTYICEVAD 2706

ZZZZZZDTYICEVEA 6345

ZZZZZZDTYICEVED 5564

EEVQLLVFGLTANSD 18582

AEVQLLVFGLTANSD 16220

EAVQLLVFGLTANSD 14220

EEAQLLVFGLTANSD 18124

EEVALLVFGLTANSD 10890

EEVQALVFGLTANSD 11258

EEVQLAVFGLTANSD 11954

EEVQLLAFGLTANSD 13317

EEVQLLVAGLTANSD 9573

EEVQLLVFALTANSD 19348

EEVQLLVFGATANSD 10408

EEVQLLVFGLAANSD 19973 EEVQLLVFGLTTNSD 20100 EEVQLLVFGLTAASD 19390 EEVQLLVFGLTANAD 17684 EEVQLLVFGLTANSA 18227 EEVQLLVFGLTANSD 19738 EEVQLLVFGLTANSD 21338 AEVQLLVFGLTANSD 14590 EAVQLLVFGLTANSD 13213 EEAQLLVFGLTANSD 16296 EEVALLVFGLTANSD 13415 EEVQALVFGLTANSD 12603 EEVQLAVFGLTANSD 13690 EEVQLLAFGLTANSD 16286 EEVQLLVAGLTANSD 11480 EEVQLLVFALTANSD 18254 EEVQLLVFGATANSD 19978 EEVQLLVFGLAANSD 18863 EEVQLLVFGLTTNSD 20021 EEVQLLVFGLTAASD 19200 EEVQLLVFGLTANAD 17928 EEVQLLVFGLTANSA 22206 EEVQLLVFGLTANSD 18721 THLLQGQSLTLTLES 7756 AHLLQGQSLTLTLES 8602 TALLQGQSLTLTLES 6931 THALQGQSLTLTLES 7683 THLAQGQSLTLTLES 7701 THLLAGQSLTLTLES 4578 THLLQAQSLTLTLES 8471 THLLQGASLTLTLES 4238 THLLQGQALTLTLES 8659 THLLQGQSATLTLES 4430 THLLQGQSLALTLES 8158 THLLQGQSLTATLES 4380 THLLQGQSLTLALES 11699 THLLQGQSLTLTAES 862 THLLQGQSLTLTLAS 2596 THLLQGQSLTLTLEA 5849 THLLQGQSLTLTLES 6545 THLLQGQSLTLTLES 4787 AHLLQGQSLTLTLES 5826 TALLQGQSLTLTLES 5012 THALQGQSLTLTLES 5059 THLAQGQSLTLTLES 5120 THLLAGQSLTLTLES 2956 THLLQAQSLTLTLES 6393 THLLQGASLTLTLES 1933 THLLQGQALTLTLES 5151 THLLQGQSATLTLES 1391 THLLQGQSLALTLES 4749 THLLQGQSLTATLES 813 THLLQGQSLTLALES 8147 THLLQGQSLTLTAES 797 THLLQGQSLTLTLAS 2193 THLLQGQSLTLTLEA 7984 THLLQGQSLTLTLES 5947 Empty (control) 569

Panel G PEPTIDE COUNTS

GEQVEFSFPLAFTVE 20691 AEQVEFSFPLAFTVE 18546 GAQVEFSFPLAFTVE 17733 GEAVEFSFPLAFTVE 17500 GEQAEFSFPLAFTVE 14764 GEQVAFSFPLAFTVE 16668 GEQVEASFPLAFTVE 6793 GEQVEFAFPLAFTVE 21681 GEQVEFSAPLAFTVE 7767 GEQVEFSFALAFTVE 20480 GEQVEFSFPAAFTVE 10024 GEQVEFSFPLTFTVE 17397 GEQVEFSFPLAATVE 10130 GEQVEFSFPLAFAVE 20627 GEQVEFSFPLAFTAE 18797 GEQVEFSFPLAFTVA 18371 GEQVEFSFPLAFTVE 17662 GEQVEFSFPLAFTVE 19190 AEQVEFSFPLAFTVE 18042 GAQVEFSFPLAFTVE 18079 GEAVEFSFPLAFTVE 19756 GEQAEFSFPLAFTVE 13000 GEQVAFSFPLAFTVE 13930 GEQVEASFPLAFTVE 6533 GEQVEFAFPLAFTVE 20072 GEQVEFSAPLAFTVE 7378 GEQVEFSFALAFTVE 19480 GEQVEFSFPAAFTVE 10589 GEQVEFSFPLTFTVE 18318 GEQVEFSFPLAATVE 9572 GEQVEFSFPLAFAVE 19516 GEQVEFSFPLAFTAE 16765 GEQVEFSFPLAFTVA 18187 GEQVEFSFPLAFTVE 18219 ZZZZZZDTYICEVED 5017 ZZZZZZDTYICEVEZ 5421 ZZZZZZDTYICEVZZ 2166 ZZZZZZDTYICEZZZ 922 ZZZZZZDTYIZZZZZ 564 ZZZZZZZTYICEVED 3031 EEVQLLVFGLTANSD 23357 EEVQLLVFGLTANSZ 15808 EEVQLLVFGLTANZZ 16496 EEVQLLVFGLTAZZZ 14097 EEVQLLVFGLTZZZZ 16473 EEVQLLVFGLZZZZZ 10516 EEVQLLVFGZZZZZZ 10372 EEVQLLVFZZZZZZZ 7333 EEVQLLVZZZZZZZZ 1098 ZEVQLLVFGLTANSD 16716 ZZVQLLVFGLTANSD 5281 ZZZQLLVFGLTANSD 4310 ZZZZLLVFGLTANSD 1026 ZZZZZLVFGLTANSD 664 ZZZZZZVFGLTANSD 779 ZZZZZZZFGLTANSD 760 ZZZZZZZZGLTANSD 657 EEVQLLVFGLTANSD 18040 THLLQGQSLTLTLES 10850 THLLQGQSLTLTLEZ^' 10269 THLLQGQSLTLTLZZ 4668 THLLQGQSLTLTZZZ 908 THLLQGQSLTLZZZZ 844 THLLQGQSLTZZZZZ 475 THLLQGQSLZZZZZZ 548 THLLQGQSZZZZZZZ 570 THLLQGQZZZZZZZZ 442 ZHLLQGQSLTLTLES 11445 ZZLLQGQSLTLTLES 11631 ZZZLQGQSLTLTLES 7993 ZZZZQGQSLTLTLES 6887 ZZZZZGQSLTLTLES 3305 ZZZZZZQSLTLTLES 4453 ZZZZZZZSLTLTLES 1086 ZZZZZZZZLTLTLES 1201 THLLQGQSLTLTLES 9756 GEQVEFSFPLAFTVE 18856 GEQVEFSFPLAFTVZ 16222 GEQVEFSFPLAFTZZ 12535 GEQVEFSFPLAFZZZ 11384 GEQVEFSFPLAZZZZ 5846 GEQVEFSFPLZZZZZ 4749 GEQVEFSFPZZZZZZ 2208 GEQVEFSFZZZZZZZ 3277 GEQVEFSZZZZZZZZ 742 ZEQVEFSFPLAFTVE 19736 ZZQVEFSFPLAFTVE 18684 ZZZVEFSFPLAFTVE 12892 ZZZZEFSFPLAFTVE 12166 ZZZZZFSFPLAFTVE 2134 ZZZZZZSFPLAFTVE 1454 ZZZZZZZFPLAFTVE 1391^' ZZZZZZZZPLAFTVE 1489 GEQVEFSFPLAFTVE 18867 empty (control) 580

Example 11

This example characterizes CD4 receptor sequences found to have HIV gpl20 binding activity in screening tests. Panel A displays information obtained from sequential replacement of amino acid residues by alaninyl residues. In panel A, a (+) signifies a decrease in binding affinity whereas a (>) indicates that replacement of the residue by an alaninyl residue yields an increase in binding affinity. Sequences are shown with amino- ter inus at the top and the carboxyl-terminus at the bottom. Right and left sides are from independent assays.

Panel A.

Panel B indicates the effect on binding affinity when successive amino acid residues are deleted, either from the amino-terminus (right side-symbols) or the carboxyl- terminus from the bottom (left side-symbol) . A (+) signifies a decrease in binding affinity, and the underlined residues indicate which residue was the last residue to be serially deleted.

Panel B.

All publications cited herein are hereby incorporated by reference to the same extent as if each publication were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

While this invention has been described with an emphasis upon preferred embodiments, it will be obvious to those of ordinary skill in the art that variations of the preferred embodiments can be used and that it is intended that the invention can be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications- encompassed within the spirit and scope of the invention as defined by the following claims.

Claims

What is claimed is:

1. A polypeptide comprising the amino acid sequence YDIXYYXXE, wherein X is any synthetic or naturally occurring amino acid residue, such that the polypeptide binds HIV gpl20 under physiological conditions, and wherein said .polypeptide comprises less than about 100 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human CCR5 chemokine receptor.

2. The polypeptide of claim 1, which comprises less than about 50 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human CCR5 chemokine receptor.

3. The polypeptide of claim 2, which comprises less than about 25 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human CCR5 chemokine receptor.

4. The polypeptide of claim 3, which comprises less than about 13 amino acids that are identical to or substantially identical to the amino acid sequence of the human CCR5 chemokine receptor.

5. The polypeptide of claim 4, which consists essentially of YDIXYYXXE.

6. The polypeptide of any of claims 1-5, which comprises the amino acid sequence YDIN*YYT*S*E, wherein N* is asparaginyl or a synthetic or naturally occurring substitute therefor, T* is threoninyl or a synthetic or naturally occurring substitute therefor, and S* is serinyl or a synthetic or naturally occurring substitute therefor.

7. The polypeptide of claim 6, wherein N* is asparaginyl, T* is threoninyl, and S* is serinyl.

8. The polypeptide of any of claims 1-6, comprising the amino acid sequence M*D*YQ*V*S*SP*IYDIN*YYT*S*E, wherein each letter indicates the standard amino acid residue designated by that letter, and a letter followed directly by an * indicates that any synthetic or naturally occurring amino acid can occupy that position.

9. The polypeptide of claim 8, wherein said letter followed directly by an * indicates the amino acid residue represented by the letter or a synthetic or naturally occurring conservative or neutral amino acid substitution therefor.

10. The polypeptide of claim 9, wherein said amino acid sequence is MDYQVSSPIYDINYYTSE.

11. A polypeptide comprising the amino acid sequence XEXIXIYXXXNYXXX, wherein X is any synthetic or naturally occurring amino acid, such that the polypeptide binds HIV gpl20 under physiological conditions, and wherein said polypeptide less than about 100 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human CXCR4 chemokine receptor.

12. The polypeptide of claim 11, which comprises less than about 50 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human CXCR4 chemokine receptor.

13. The polypeptide of claim 11, which comprises less than 25 contigous amino acids that are identical to or substantially identical to the amino acid sequence of the human CXCR4 chemokine receptor.

14. The polypeptide of claim 13 , which consists essentially of EXIXIYXXXNY.

15. The polypeptide of any of claims 11-14, which comprises the amino acid sequence M*EG*IS*IYT*S*D*NYT*E*E*, wherein each letter indicates the standard amino acid residue designated by that letter, and each letter followed directly by an * indicates the amino acid residue represented by the letter or a synthetic or naturally occurring conservative or neutral amino acid substitution therefor.

16. The polypeptide of claim 15, wherein said amino acid sequence M*EG*IS*IYT*S*D*NYT*E*E* is M*EGISIYTSDNYT*E*E* .

17. A polypeptide comprising the amino acid sequence EHQAFLQFS, such that the polypeptide binds with HIV gpl20 under physiological conditions and wherein said polypeptide comprises less than about 100 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human STRL33 chemokine receptor.

18. The polypeptide of claim 17, which comprises less than about 50 contiguous amino acid that are identical to or substantially identical to the amino acid sequence of the human STRL33 chemokine receptor.

19. The polypeptide of claim 18, which comprises less than about 25 contiguous amino acids that are identical to or substantially identical to the amino acid sequence of the human STRL33 chemokine receptor.

20. The polypeptide of claim 19, which consists essentially of the sequence EHQAFLQFS.

21. A polypeptide comprising at least a portion or all of an amino acid sequence selected from the group consisting of LPPLYSLVFIFGFVGNML, QWDFGNTMCQLLTGLYFIGFFS, SQYQFWKNFQTLKIVILG, APYNIVLLLNTFQEFFGLNNCS, and

YAFVGEKFRNYLLVFFQK, wherein the polypeptide binds with HIV gpl20 under physiological conditions and comprises less than about 100 amino acid residues that are identical to or substantially identical to the amino acid sequence of the human CCR5 chemokine receptor.

22. A polypeptide comprising at least a portion or all of an amino acid sequence selected from the group consisting of LLLTIPDFIFANVSEADD (165-182) , WFQFQHIMVGLILPGIV (197-214) , and IDSFILLEIIKQGCEFEN (261-278) , wherein the polypeptide binds with HIV gpl20 under physiological conditions and comprises less than about 100 amino acid residues that are identical to or substantially identical to the amino acid sequence of the human CXCR4 chemokine receptor.

23. A polypeptide comprising at least a portion or all of an amino acid sequence selected from the group consisting of LVISIFYHKLQSLTDVFL (53-70), PFWAYAGIHEWVFGQVMC (85-102) , EAISTWLATQMTLGFFL (185- 202), LTMIVCYSVIIKTLLHAG (205-222),

MAVFLLTQMPFNLMKFIRSTHW (237-258) , HWEYYAMTSFHYTIMVTE (257-274) , ACLNPVLYAFVSLKFRKN (281-298) and SKTFSASHNVEATSMFQL (325-342) , wherein the polypeptide binds with HIV gpl20 under physiological conditions and comprises less than about 100 amino acid residues that are identical to or substantially identical to the amino acid sequence of the human STRL33 chemokine receptor.

24. A polypeptide comprising at least a portion of or all of an amino acid sequence selected from the group consisting of DTYICEVED, EEVQLLVFGLTANSD, THLLQGQSLTLTLES, and GEQVEFSFPLAFTVE, wherein the polypeptide binds with HIV gpl20 under physiological conditions and wherein the polypeptide comprises less than about 100 amino acids that are identical to or substantially identical to the amino acid sequence of the human CD4 cell-surface protein.

25. A polypeptide of any of claims 21-24, which comprises all of the amino acid sequence and 0 to about 6 conservative or neutral amino acid substitutions.

26. The polypeptide of claim 25, comprising 0 amino acid substitutions.

27. The polypeptide of any of claims 21-26, which comprises less than about 50 amino acids that are identical to or substantially identical to a protein that naturally has the amino acid sequence.

28. The polypeptide of any of claims 21-26, which comprises less than about 25 amino acids that are identical to or substantially identical to a protein that naturally has the amino acid sequence.

29. The polypeptide of any of claims 1-28, wherein said polypeptide further comprises a pharmaceutically acceptable substituent.

30. A composition comprising the polypeptide of any of claims 1-28, and a carrier.

31. A nucleic acid encoding the polypeptide of any of claims 1-28, wherein said nucleic acid can be expressed in a cell.

32. The nucleic acid of claim 31, further comprising a nucleic acid sequence that encodes a signal sequence, wherein said signal sequence is translated as a fusion protein with the polypeptide to form a signal sequence-polypeptide fusion, and wherein said signal sequence can cause secretion of at least the polypeptide out of a cell in which the nucleic acid is expressed.

33. A vector comprising the nucleic acid of claim 31 or 32.

34. A method of making an antibody, which method comprises administering an immunogenic amount of a polypeptide of any of claims 1-28 or a nucleic acid of any of claims 31 or 33 to an animal.

35. A method of prophylactically or therapeutically treating HIV infection in a mammal in need thereof, which method comprises administering to said mammal an effective amount of a polypeptide of any of claims 1-28, a nucleic acid of any of claims 31-33, or an anti- antibody to a polypeptide of any of claims 1-28.

36. A method of making an antibody that binds to a gpl20 envelope protein of a human immunodeficiency virus-1 (HIV-1) , said method comprising:

(a) labeling a polypeptide of any of claims 1-28 to obtain a labeled compound,

(b) providing a library of synthetic peptides, wherein said library consists of a multiplicity of synthetically-produced polypeptides that are homologous to a continuous region of an HIV-1 gpl20 envelope protein, wherein each polypeptide of said library is substantially isolated from every other polypeptide of said library and is located in a known position, (c) individually contacting each polypeptide with said labeled compound such that a portion of the labeled compound can bind with the polypeptide, thereby producing a bound population of each polypeptide and an unbound population of each polypeptide, (d) removing substantially all of the unbound labeled compound from the position occupied by each polypeptide,

(e) measuring the amount of labeled compound that remains co-localized with each polypeptide, to determine the quantity of labeled compound bound by each polypeptide,

(f) evaluating the amount of labeled compound bound by each polypeptide to identify a portion of the HIV-1 gpl20 envelope protein that binds to an (HIV-1) -receptor selected from the group consisting of CCR5, CXCR4, STRL33, and CD4,

(g) providing an immunizing compound comprising a polypeptide comprising an amino acid sequence that is homologous to said portion of the HIV-1 gpl20 envelope protein,

(h) inserting an immunogenic quantity of said immunizing compound into an animal to cause said animal to produce an, antibody that binds with said portion of the HIV-1 gpl20 envelope protein.

37. The method of claim 36, wherein said labeled compound comprises a moiety selected from the group consisting of a radioactive atom, an enzyme, a polyhistidinyl moiety, and an antigen that is specifically recognized by a standard antibody.

38. The method of claim 36 or 37 wherein said library consists of a multiplicity of synthetically- produced polypeptides that are identical to a continuous region of an HIV-1 gpl20 envelope protein.

39. The method of any of claims 36-38, wherein said polypeptides contain at least about 6 amino acid residues and no more than about 45 amino acid residues.

40. The method of claim 39,. wherein said polypeptides contain no more than about 30 amino acid residues .

41. The method of any of claims 36-40, wherein said library comprises a multiplicity of polypeptides of identical lengths.

42. The method of any of claims 36-41, wherein said library comprises a multiplicity of polypeptides that are homologous to a region of the HIV-1 gpl20 envelope protein and have an offset of n amino acid residues, wherein n is an integer of at least l and is not greater than the product of length of the longest polypeptide measured in amino acid residues and 1.5.

43. The method of claim 42, wherein said offset is not greater than the product of length of the longest polypeptide measured in amino acid residues and 1.0.

44. The method of claim 42, wherein said offset is not greater than the product of length of the longest polypeptide measured in amino acid residues and 0.5.

45. The method of claim 42, wherein said offset is not greater than 30.

46. The method of claim 42, wherein said offset is not greater than 15.

47. The method of claim 42, wherein said offset is not greater than 4.

48. The method of any of claims 36-47, wherein each polypeptide is bound to a solid support and is located in a vessel that enables each polypeptide to be covered in a liquid that does not contact any other oligonucleotide of the library.

49. The method of claim 48, wherein each polypeptide is bound to a bead in a vessel or is bound to the well of a multi-well assay plate.

50. The method of claim 36, wherein said step of removing substantially all of the unbound labeled compound comprises the additional steps of (i) removing a liquid containing said unbound labeled compound from a solid substrate to which an polypeptide of the library is bound, (ii) applying a quantity of wash-liquid to said solid substrate that is sufficient to cover any portion of said solid substrate or a vessel containing said solid substrate that has been contacted by said labeled compound, and (iii) removing said wash-liquid.

51. The method of any of claims 36-50, wherein said immunizing compound comprises an adjuvant or wherein said polypeptide comprising an amino acid sequence that is homologous to said portion of the HIV gpl2Q envelope protein is conjugated to a known immunogen.

52. The method of any of claims 36-51, wherein said method is performed in a mammal belonging to a group selected from- the group consisting of rodents, canines, felines, and ruminants.

53. The immunizing compound of step (g) of the method of any of claims 36-52.

54. An antibody produced by the method of any of claims 36-53.

55. A method of removing HIV from a bodily fluid of a mammal", which method comprises extra-corporeally contacting said bodily fluid with a solid support to which is attached a polypeptide of any of claims 1-28 or an anti-antibody to a polypeptide of any of claims 1-78, or the antibody of claim 54.