NZ223494A

NZ223494A - Peptides, nucleic acids and oligonucleotides related to sequences of hiv-2 and siv-1

Info

Publication number: NZ223494A
Application number: NZ223494A
Authority: NZ
Inventors: Marc Alizon; Luc Montagnier; Denise Guetard; Francois Clavel; Pierre Sonigo; Mireille Guyader; Pierre Tiollais; Lisa Chakrabarti; Ronald Desrosiers
Original assignee: Pasteur Institut
Priority date: 1987-02-11
Filing date: 1988-02-11
Publication date: 1991-06-25
Also published as: CA1341634C; OA08716A; DE3856595D1; KR0135885B1; IE990955A1; EP0750041A3; MX169441B; ATE417108T1; EP0750041B1; EP0750041A2; KR890700603A; CA1341520C

Abstract

New peptides (I) have immunological properties in common with those of the peptide skeleton of the envelope protein of HIV-2 and also have a peptide structure in common with that of SIV (simian immunodeficiency virus)-7 glycoprotein. Also new are (i) a sequence of 9600 nucleotides corresponding to the SIV genome (reproduced in the specification, together with derived amino acid sequence of viral proteins gag, pol, env, Q, X, R, tat, art and F genes) and its fragments; (2) recombinant DNA contg. all or part of the CDNA from this sequence inserted into a vector, and (3) antigenic and immunogenic conjugates contg. (I).

Description

<div id="description" class="application article clearfix"> 2234* Priority Date(s): II 2 Complete Specification Filed: v.'.'rr.Hr. Sol .N3.3 j ^'4* * Publication Date: P.O. Journal. No: . Parents Forn No. 5 NEW ZEALAND PATENTS ACT 1953 COMPLETE SPECIFICATION PEPTIDES CAPABLE OF BEING RECOGNIZED BY ANTIBODIES INDUCED AGAINST RETROVIRUSES OF HUMAN IMMUNODEFICIENCY (HIV VIRUSES), THEIR USES FOR THE DIAGNOSIS OF INFECTIONS DUE TO SOME OF THESE VIRUSES AND, WHERE APPROPRIATE FOR VACCINATION AGAINST AIDS 7/We, INSTITUT PASTEUR, a French Public establishment of 25-28 rue du Dr. Roux, 75724 Paris, Cedex 15, France, hereby declare the invention, for which X/ve pray that a patent may be granted to jj£/us, and the method by which it is to be performed, to be particularly described in and by the following statement: (followed by Page la) § - la - o c * r s ^ J d ^ o a;; 4 15 20 25 30 35 PEPTIDES CAPABLE OF BEING RECOGNIZED BY ANTIBODIES INDUCED AGAINST RETROVIRUSES OF HUMAN IMMUNODEFICIENCY (HIV VIRUSES), THEIR USES FOR THE DIAGNOSIS OF INFECTIONS DUE TO SOME OF THESE VIRUSES AND, WHERE APPROPRIATE FOR VACCINATION AGAINST AIDS. The present invention relates to peptides having immunological, even immunogenic, properties in common with antigens which can be obtained in a purified form from viruses capable of causing lymphadenopathies which, in man, may subsequently degenerate into the acquired inmuno-10 deficiency syndrome (AIDS). The invention relates in particular to antigenic pept'ides capable of being recognired by antibodies induced in man by viruses designated by the abbreviation HIV according to the nomenclature defined in NAT'JRZ. The invention also relates to peptides having immunogenic properties or which are capable of being rendered immunogenic in vivo, this imr.unogenicity manifesting itself by the induction in vivo of antibodies recognizing antigens characteristic of the viruses HIV-2 and even, at least in the ca6e of certain of these peptides, antigens derived from HIV-1. The invention relates in addition to uses of these peptides for the mass production of compositions for the in vitro diagnosis in man of the latent existence of certain forms of AIDS and, in the case of some of the AIDS viruses, for the production of immunogenic compositions and of compositions to be used as vaccines against the HIV retroviruses. Similarly, the invention relates to the uses for the same purpose of the antibodies capable of being induced in vivo by the immunogenic peptides or peptides made immunogenic and, in the case of some of these antibodies, to their uses for the production of medicinal active principles against these human AIDS. The invention also relates to the utilization of some of these peptides in procedures for the in vitro diagnosis in man of some forms of AIDS as well as their use for making up diagnostic "kits". The first retrovirus designated LAV-1 or HIV-1 was isolated and described in nz 209556. This virus has also been described by F. Barre Sinoussi et al. in Science, 220 No. 45-99, 20 pages S6S-S71. (followed by page 2} 1 5 10 15 20 25 30 35 Variants of this HIV-1 virus designated as LAV ELI and LAV MAL have also been isolated, characterized and described in NZ 209556. • The HIV-1 viruses and their variants possess the following properties: - their preferred targets are the human Leu3 cells (or T4 lymphocytes) and the "immortalized" cells derived from them. - they have a reverse transcriptase activity requiring the presence of Mg T ions and exhibit a marked activity toward poly(adenylate-oligo-deoxythymidylase) poly(A)-oligo(dT)12-18). - they have a density of 1.16 to 1.17 on a sucrose gradient, - they have an average diameter of 139 nanometers and 'a nucleus with an average diar.eter of 41 nanometers, - the lysates of these viruses contain a protein p25 (core protein) which immunologically does not cross-react with protein p24 of HTLV-1, - they contain a protein p^2 as a constituent of their envelope, - they also contain a glycoprotein gpllO of molecular weight 110,000 in their envelope. The isolation and characterization of retroviruses belonging to a distinct class which are immunologically related to those just mentioned only to a limited extent have been described in the NZ patent application No. 219024 These retroviruses which have been grouped together under the designation HIV-2 were isolated from several African patients exhibiting symptoms of a lymphadenopathy or an AIDS. The retroviruses of the HIV-2 type like the retroviruses of the HIV-1 type are characterized by a tropism for the human T4 lymphocytes and by a cytopathogenic effect on these lymphocytes as a result of multiplying within them, thus giving rise either to generalized and persistent polyadenopathies or an AIDS. More generally, the retroviruses purified by HIV-2 possess the following properties: - the preferred target of the HIV-2 retroviruses is constituted by~femnan Leu3 cells (or T4 lymphocytes) and for "immortalized" cells derived from these T4 lymphocytes; - they are cytotoxic for human T4 lymphocytes 22 3 4 9 i 5 0 5 0 - they have a reverse transcriptase activity requiring the presence of 2+ Mg ions and exhibit a marked activity for poly(adenylate-oligodeoxy-thymidylase) (poly(A)-oligo(dT) 12-18); - they have a density of 1.16 in a sucrose gradient; - they have a mean diameter of 140 nanometers and a nucleus having a mean diameter of 41 nanometers; - they can be cultured in permanent cell lines of the HUT type or those expressing the T4 protein; - they do not infect T8 lymphocytes; - the lysates of these viruses contain a protein p26 which immunologically does not cross-react with the protein p24 of the virus HTLV-I or of the virus HTLV-II; - in addition, these lysates contain a protein pl6 which is not recognized immunologically by the protein pl9 of HTLV-I or of HTLV-II in radioimmuno-precipitation assays. - in addition, they contain a glycoprotein having a molecular weight of the order of 130,000-140,000 in their envelope which immunologically does not cross-react with the gpllO of the HIV-1, but which, in contrast, does cross-react immunologically with the glycoprotein gpl40 of the envelope of STLV-III (a virus isolated from monkeys); - these lysates also contain antigens which can be labelled with 35 S-cysteine, the molecular weights of which range between 32,000 and 42,000-45,000: they include in particular an antigen having a molecular weight of the order of 36,000 and an antigen having the molecular weight of the order of 42,000, one of these antigens (p36 or p42) probably constituting a transmembrane glycoprotein of the virus HIV-2; - the genomic SNA of the HIV-2 does not hybridize with the genomic RNA of HIV-1 under stringent conditions; - under non-stringent conditions, the genomic RNA of HIV-2 hybridizes neither with the env gene and the LTR which is next to it of HIV-1 nor with the sequences of the pol region of the genome of HIV-1; - under non-stringent conditions, it hybridizes weakly with nucleotide sequences of the region of HIV-1. Another retrovirus designated SIV-1, this designation replacing the earlier one of STLV III, was isolated from the rhesus macaque monkey (M. D. Daniel et al., Science 228, 1201.(1985); N. L. Letwin et al., - A - 22 3 4 9 v-V c :o 15 20 25 30 35 Science 230, 71 (1985) under the name "STLV-IIImac"). Another retrovirus, designated "STLV-III^^" (or SIVAGM) was isolated from wild green monkeys. However, in contrast to the virus present in the rhesus macaque monkeys, the presence of "STLV-III, _" does not seem AGM to induce a disease of the AIDS type in the African green monkey. A strain of the SIV-lmac retrovirus was deposited with the C^CM on 7th February 1986 under the No. 1-521. Studies have shown that the retrovirus SIV-1 contains some proteins which are immunologically related to structural proteins or glycoproteins which can be isolated from HIV-2 under similar conditions. This retrovirus SIV-1, the infectious nature of which has been observed in monkeys, has been designated as STLV-III by the research scientists who have isolated it (literature references cited above). For semantic convenience, these viruses will be referred to in the following discussion only by the expression SIV (the expression SIV is an abbreviation for "Simian Immunodeficiency Virus") which may be followed by an abbreviation indicating the species of monkey from which it was derived, for example MAC (or mac) for the macaque or AGM for the African green monkey. Using the same methods as those mentioned above, it has been observed that the SIV-1 mac also contains: - a major nuclear protein p27 having a molecular weight of the order of 27 kilodaltons, - a major glycoprotein, gpl40, in its envelope, - a protein p32, probably transmembrane, which is hardly observed in 35 RIPA when the virus has been labelled beforehand with S—cysteine, but which can be observed in the form of tiroad bands on Western blots. More precise studies have been carried out on the HIV-2 and SIV viruses. The continuation of the study of the HTV-2 retroviruses has also led to the preparation of complementary DNA sequences (cDNA) of the RNAs of their genomes. The complete nucleotide sequence of the cDNA of a representative retrovirus of the HIV-2 class (HIV-2 ROD) was deposited on 21/02/1986 with the CNCM under No. 1-522 and the reference LAV-II ROD) . This nucleotide sequence and the open-reading frames which it contains are indicated in the Figure 1A. 22 3 4 9 4 Furthermore, continuation of the study of other retroviruses has also made it possible to obtain their complete nucleotide sequences. This is the case in particular for the cDNA derived from the genomic RNA of SIV. The cloning and sequencing of the SIV-lmac virus which led to the elucidation of its nucleotide sequence were carried out under the following conditions: The DNA of HUT 78 cells infected with the SIV virus (isolate STLV-III mac 142-83 described by Daniel et al. (1985) Science, 228, p. 1201-1204, partiallv digested by the restriction enzyme Sau3A was cloned at the "BamHI site of the bacteriophage vector lambda ELBL3 in order to constitute a genomic library.The 2million recombinant phages of the genomic library thujs constituted were screened in situ under P3 conditions of security with the aid of sequences of the HIV-2 virus, derived from clones of lambda-R0D4, lambda-R0D35 and E2 (Clavel et al., 1986, Nature, 324, p. 691.) and nick-translated. Hybridization was carried out in 5 x SSC at 50°C and washings in 2 x SSC at 50°C. A single clone containing all of the viral sequences was obtained. This clone was designated as lambda-SIV-1. The insert of the phage lambda-SIV-1 measures a total of 16.5 kb and includes an integrated provirus which lacks only the first 250 bases of the leftside of LTR whereas the right-side of LTR is complete. The integrated provirus was sequenced by the dideoxynucleotide method after subcloning of random fragments in the phage M13mp8. 300 subclones were analyzed. Fragments of cDNA derived from the clone lambda-SIV-1 inserted into plasmids pSIV-1.1 and pSIV-1.2 were deposited with the CNCM on 15 th April 1987 under the numbers 1-658 (pSIV-1.1) and 1-659 (pSIV-1.2). The results are mentioned in the figures described below. Figure IB presents the nucleotide sequence of the viral genome of SIV and the sequences which have been deduced from it for the-viral proteins corresponding to the gene products gag, pol, env, Q, X, R, tat, art, F. The Figures 3 to 11 and the Figure 1C present comparisons between HIV-2 and SIVmac of the theoretical products of the viral genes and the LTR CXSIV-1). 22 3 4 9 4 i 5 10 15 20 25 30 35 The invention also relates to fragments of cDNA deduced from the cDNA derived from the entire genome of SIV-1, these fragments containing one or more sequences derived from the complete sequence of cDNA and which code for interesting peptides of the invention. These sequences are indicated in Figure IB and that relating to the LTR sequence of the virus in Figure 1C. The nucleotide sequences of the cDNA of SIV were matched with the nucleotide sequences of the virus HIV-2 ROD with respect to the LTR sequence (Figure 1C). This type of match which is made for the entire genome by comparing Figure IB with Figures 3 to 11 makes it possible to locate or deduce the nucleotide sequences of the two viruses which have common essential structural elements. The invention naturally also relates to the use of the cDNAs derived from SIV or their fragments (or recombinants containing them) both as probes for diagnosing the presence or absence of HIV-2 viruses in serum samples or other liquids or biological tissues obtained from patients suspected to be carriers of the HIV-2 virus. These probes are preferably also labelled (radioactive, enzymatic, fluorescent markers, etc). Probes of particular interest for the application of the diagnostic procedure for the HIV-II virus or a variant of HIV-2 may be characterized in that they contain the totality or a fraction of the cDNA complementary to the gene of the SIV virus or particularly the recombinant fragments contained in various clones. The probes used in this diagnostic procedure for the HIV-2 virus and in the diagnostic kits are in no way limited to the probes described above. On the contrary, they comprise all of the nucleotide sequences derived from the genome of the SIV virus, of a variant of SIV or of a virus closely related in structure as long as they make it possible'to detect in the biological fluids of persons suspected of developing an AIDS,antibodies directed against a HIV-2 or a virus which i? closely related to it. The detection may be carried out by any known method. It may comprise the placing in contact of these probes either with the nucleic acids obtained from cells contained in these sera or other biological fluids, for example cerebrospinal fluid, saliva, etc ... It may also 223494 '• *1 4 ? v. % o 15 20 25 30 35 involve placing these probes in contact with these fluids themselves once their nucleic acids have been made accessible to hybridization with these probes under conditions permitting hybridization between these probes and these nucleic acids to occur. The final step of in vitro diagnosis thus comprises the detection of the hybridization which may have occurred. The above-mentioned diagnostic procedure involving hybridization reactions may also be carried out with the aid of mixtures of probes derived from a HIV-2 and a SIV-1 or a HIV-1, a HIV-2 and a SIV, respective!" if it is not necessary to distinguish between the type of virus searched. Generally speaking, the diagnostic procedure for the presence or absence of the HIV-2 virus or a variant in serum samples or other liquids or tissues obtained from patients suspected of being carriers of the HIV-2 virus comprise the following steps: 1) at least one hybridization step conducted under stringent conditions by placing in contact the DMA of the cells of the sample taken from the suspected patient with one of the above-mentioned labelled probes on an appropriate membrane, 2) the washing of the said membrane with a solution which maintains these stringent conditions of hybridisation, 3) the detection of the presence or absence of the HIV-2 virus by a method of immunodetection. In another preferred embodiment of the procedure according to the invention the_ above-mentioned hybridization is conducted under non-stringent conditions and the washing of the membrane is carried out under conditions suited to those of the hybridization. It will be obvious that the invention relates to the nucleic acids corresponding to sequences located in analogous regions of variants of SIV as well as to all of the nucleic acids in which modification would make it possible to take advantage of the degeneracy of the genetic code. The comparative studies which have also enabled results -to be obtained relating to the core proteins, hereafter termed "gag proteins" and to proteins of the envelope, hereafter referred to as "env proteins", have also been reported in the NZ patent application No. alreadv mentioned. These results show that the core pr< proteins) in HIV-2 exhibit less marked differences frow'VHose of the"3s8,V-l - 8 - 22 3 4 9 4 i 5 10 15 20 25 30 35 viruses than the proteins of the envelope (env proteins). Overall, the env proteins of HIV-2 have been found to be immunologically related with the corresponding env proteins of the HIV-1 viruses to only a very slight degree or not at all. On the other hand, comparative studies on the structures of the cDNA sequences of the HIV-2 and SIV viruses have demonstrated certain common properties which make their appearance at the level of the proteins. Overall, the proteins of HIV-2 and SIV-1 show a considerable degree of immunological relatedness. The major glycoprotein of the envelope of HIV-2 has been shown to be more closely related immunologically to the major glycoprotein of the envelope ofSIV than to the major glycoprotein of the envelope of HIV-1. These observations can be affirmed not only with regard to the molecular weights: 130-140 kilodaltons for the major glycoproteins of HIV-2 and SIV compared with about 110 kilodaltons for the major glycoprotein of the envelope of HIV-1, but also with respect to their immunological properties since sera taken from patients infected with HIV-2, and more particularly antibodies formed against the gpl40 of HIV-2 recognize the gpl40 of SIV-lmac, whereas in similar assays the same sera and the same antibodies of HIV-2 do not recognize the gpllO of HIV-1. However, the anti-HIV-1 sera which have never reacted with the gpl40 of HIV-2 precipitatt 35 a protein of 26 Kdal labelled with S-cysteine contained in the extracts of HIV-2. The major core protein of HIV-2 seems to possess an average molecular weight (about 26,000) intermediate between that of the p25 of HIV-1 and the p27 of SIV. These observations derive from assays carried out with viral extracts obtained from a HIV-2 isolated from one of the patients mentioned above. Similar results have been obtained with viral extracts of a HIV-2 isolated from another patient. More extensive studies have led the inventors to recognize an initial class of peptides having sequences of amino acids which are either identical or very similar to sequences contained in the interior of the structures of the gag and env proteins of HIV-2 or SIV, or even HIV-1. These 22 3 4 9 4 1 peptides are particularly useful for the diagnosis of an infection by the HIV-2 virus or one of its variants in man. In this respect, the present invention also relates to diagnostic procedures and compositions for the in vitro detection of antibodies 5 directed against a HIV-2 virus or its variants, more particularly in biological samples, particularly in the sera of patients who have become infected with the HIV-2 virus, since some of these peptides make possible a particularly fine distinction between the infections due to HIV-2 viruses and those due to HIV-1 viruses. 10 These more exhaustive studies have also led to the possibility of synthesizing immunogenic peptides or peptides capable of being made immunogenic which exhibit structural characteristics enabling them to induce in vivo the production of antibodies capable of recognizing env proteins both of HIV-1 and HIV-2 and, at least in the case of some of 15 these peptides, of binding both to HIV-1 viruses and to HIV-2 viruses, and resulting more particularly in their neutralization. The use of these latter types of peptides is thus particularly indicated for the production of active principles of vaccines against the HIV viruses, and hence against the AIDS. 20 In order to specify in the subsequent discussion the amino acid residues which constitute the peptides according to the invention, resort will be had to the code shown in the table which follows for those amino acids having an unequivocal meaning in the international nomenclature which designates each naturally occurring arm"no acid by a single letter (capital letter): M Methionine L Leucine I Isoleucine V Valine 7 Phenylalanine S Serine P Proline T Threonine A Alanine Y Tyrosine - 10 - 22 3 4 9 4 i 5 10 15 20 25 30 35 H Histidine Q Glutamine N Asparagine K Lysine D Aspartic acid E Glutamic acid C Cysteine w Tryptophan R Arginine G Glycine When, on account of its position within the chain of amino acids characteristic of a specific peptide, an amino acid can assume several meanings, it will be designated either by a dash if it can be any amino acid, or by a lower case letter when this amino acid can assume a limited number of preferred meanings, this number being, however, always higher than one. In this latter case, the possible meanings of this lower case letter will always be specified in relation to the peptide to which it belongs. In order to facilitate reading, these peptides will be designated by an abbreviation env or gag followed by a number referring to sequences of amino acids containing either in the env proteins or in the gag proteins of certain HIV-1, HIV-2 or SIV. Reference will also be made to them in the ensuing discussion. Finally, in the definitions which follow - the X groups represent either a free group or one ami dated in particular by one or two alkyl groups containing from 1 to 5 carbon atoms, or a peptide group comprising from 1 to 5 amino acids, the N-terminal amino acid of which itself possesses a free group or one amidated as previously mentioned, and - the Z groups represent either a free -OH group or an alkoxy. group contain ing an alkyl group comprising from 1 to 5 carbon atoms, or a peptide group comprising from 1 to 5 amino acids, the C-terminal amino acid of which itself possesses a free -OH group of an alkoxy group as previously indicated, the groups of from 1 to 5 amino acids contained,as appropriate, in X or Z or in both at once being such that their presence is for the - 11 - 22 3 4 9 10 15 20 25 30 most part not incompatible with the preservation of the immunological or even immunogenic, properties of the peptides which do not contain them. The peptides according to the invention which have immunological properties in common with antigens of HIV-2 and, in certain cases, also with antigens of HIV-1 or its variants are characterized in that they also have a peptide structure in common with the antigens of SIV. In an advantageous manner, these peptides normally contain a maximum of 40 amino acid residues. The following are preferred peptides: envl XRV-AIEKYL-DQA-LN-WGCAFRQVCZ env2 X-LE-AQI-QQEKNMYELQKLN Z env3 , XELGDYKLVEITPIG-APT —KR Z env4 X VTV-YGVP-WK-AT—LFCA-Z env5 X QE--L-NVTE-F—W-NZ env6 XL S-KPCVKLTPLCV—Z env7 X—N-S-IT—C-K Z env8 X-I YC-P-G-A-L-C-N-TZ env9 X A-C W— Z env 10 X-G-DPE NC-GEF-YCN NZ envl 1 x C-IKQ-I G---YZ 35 . 22 3 4 9 4 W7 / 10 15 20 25 30 35 More particularly, the invention relates to the following peptides: envl : XRV-AIEKYL-DQA-LN-WGCAFRQVCZ env2 X-LE-AQIQQEKNMYELQKLN Z env3 XELGDYKLVEITPIG-APT—KR Z env4 X VTV-YGVP-W--AT—LFCA-Z env5 X E—L-NVTE-F--W-NZ env6 XL S -KPCVKL - PLC Z env7 X N-S-I C-K Z env8 X-I YC-P-€-A-L-C-N-TZ env9 X A-C W—Z env 10 X-G-DPE NC-GEF-YC NZ envl 1 X C-I-Q-I G YZ Useful peptides corresponding to those just cited above are presented in the formulas which follow: envl XRVTAIEKYLQDQARLNSWGCAFRQVCZ, or XRVTAIEKYLKDQAQLNAWGCAFRQVCZ env2 XSLEQAQIQQEKNMYELQKLNSWZ, or XLLEEAQIQQEKNMYELQKLNSWZ env3 XELGDYKLVEITPIGFAPTKEKRYSSAHZ,or XELGDYKLVEITPIGLAPTNVKRYTTG-Z - 13 - 22 3 4 (It will be noticed that the peptides envl, env2, env3 testify to the very close relationship between HIV-2 and SIV-1. In fact, the first peptide is included in the genome of HIV-2 and the second in that of SIV-1). env4 XabcdVTVeYGVPfWogAThiLFCAjZ, in which the letters from a to j may have the following meanings: a is C, E or D b is T, K, D, N or I c is 0 or L d is Y or W e is F or Y f is T, V or A g is N or E h is I or T i is P or T j is T or S o is K or R env5 XabcoEdeLfNVTEgFhiWjNZ, in which the letters from a to j may have the following meanings: a is D or P b is D or N c is Y or P d is I, V, I or L e is T, V, E or A f is V, G or E or g is A, N, G or S h is D or K i is A or H j is u o W * 5 E 0 is Q or S env6 XLabcSdKPCVKLoPLCuefKZ, m - 14 - 22 3 4 9 4 o *1* % 10 15 20 25 30 35 in which the letters from a to f may have the following meanings: a is F or W b is E or D c is T or Q d is I or L e is A, S or T f is M or L o is T or S u is V or I env7 XabCNxSylocdCeKfghiZ, in which the letters from a to i and x and y may have the following meanings: a is N or T or I b is H or S or N c is E or Q d is S, A or C e is D or P f is H, V or D g is Y or S h is W or F i is D or E X is T or R 7 is V or A o is T or Q «?PV_8 XaIbedYCxPeGfAgLhCiNjTZ, in which the letters from a to k and x may have the following meanings: a is A or P b is R or P c is F, I or C d is R or H e is P or A f is Y or F S is L or I h is R or K - 15 - 22 3 4 9 4 o 10 15 20 : c 25 i c 30 35 I is - or N j is D or K x is A or T env9 XwabcxyAdCefghizWjkZ, in which the letters from a to k and x to z may have the following meanings: a is K or - or E b is R or - c is P or M or I d is W or H or Y e is W or N or T or R f is F or I g is K or S or N or G h is G or R or E i is - or A or T j is K or N or D or S k is D or A or N or K or E w is N, 8 or I x is R or G or K y is Q or K or R z is K or E or Q or N env 10 XaGbDPEcdefghNCiGEFjYCokxlmnNZ, in which the letters from a to n and x may have the following meanings: a is K or - or G b is S or G or - c is V or I d is A or V or T . e is Y or T or M or F f is M or H g is W or S h is T or F i is R or G - 16 - 22 3 4 9 4 r>i C ;o 15 20 25 30 35 j is L or F 0 is N or K k is M or S 1 is W or Q or K or G m is F or L n is L or F x is T or S or N envl 1 XabcdwCeloQfIxgyhizGjklYZ, in which the letters from a to 1 and w to z may have the following meanings: a is R or T or S or N b is N or I c is Y or T d is A or L or V e is H or R f is I or F g is T or M h is H or Q or A i is K or E j is R or K k is N or A 1 is V or M w is P or Q x is N or K y is W or V z is V or T or K o is K or R The structure of the antigenic peptide coded by the gene gag and designated by gag 1 is also shown below: XDCKLVLKG LGaNPT LEEMLTAZ, in which the letter a specifies M or T. It will be noticed that, generally speaking, the amino acids having an unequivocal meaning (hence represented by a capital letter in accordance with the international nomenclature) which appear in the definitions which - 17 - 11 3 4 9 4 * \ -» T^\ .■ •; c 10 15 20 25 30 precede of- the peptides according to the invention are found to correspond to identical amino acids placed in the same order in the corresponding env or gag sequences of the env or gag protein of at least one of the HIV or of SIV-1 The positions of the sequences are underlined and located within the sequences of amino acids of the env proteins of HIV-2 ROD (CNCM No. 1-532) and HIV-1 BRU (CNCM No. 1-232) shown in Figure 2. Moreover, the alignments of the amino acids of the env and gag proteins of SIV-lmac (CNCM No. 1-521) and of HIV-2 ROD are presented in Figures 3 and 4. The continuous lines which replace individual dashes at certain locations in these sequences is intended to underline the fact that certain amino acids contained in these.sequences have been deliberately omitted from the figure in order to allow identical amino acids to be aligned (in that case marked with an asterisk) or for two points to be aligned vertically in the sequences of the corresponding proteins of HIV-1 and HIV-2 on the one hand, and SIV and HIV-2 on the other. In addition to the peptides already mentioned, the invention also relates to peptides modified by insertion and/or deletion and/or substitution of one or more amino acids provided that the antigenic or immunogenic properties of the said peptides are not modified and that the recognition properties of the antigen and the antibody for these peptides are not substantially modified. In a particularly preferred embodiment the invention relates to peptides having immunological properties in common with the peptide skeleton of the glycoprotein of the envelope of the viruses of the class HIV-2, these peptides containing a number of amino acid residues not exceeding 40. These preferred peptides according to the invention have the following sequences: 35 envl RVTAIEKYLQDQARLNSWGCAFRQVC AIEKYLQDQ RVSAIEKYLKDQAQLNAWGCAFRQVC AIEKYLKDQ env2 SLEQAQIQQEKNMYELQKLNSW QIQQEKN LLEEAQIQQEKNMYELQKLNSW env3 ELGDYKLVEITPIGFAPTKEKRYSSAH YKLVEITPIGFAPTKEK ELGDYKLVEITPIGLAPTNVKRYTTG-YKLVEITPIGLAPTNVK env4 CTQYVTVFY GVPTWKNATIPLFCAT VTVFYGVPTWKNAT CIQYVTVFYGVPAWRNATIPLFCAT VTVFYGVPAWRNAT EKLWVTVYYGVPVWKEATTTLFCAS VTVYYGVPVWKEAT EDLWVTVYYGVPWKEATTTLFCAS VTVYYGVPVWKEAT DNLWVT\frYYGVPVWKEATTTLFCAS VTVYYGVPVWKEAT env5 DDYQEITL-NVTEAFDAWNN L-NVTEAF DDYSELAL-NVTESFDAWEN L-NVTESF PNPQEVVLVNVTENFNMWKN LVNVTENF PNPQEIELENVTEGFNMWKN LENVTEGF PNPQEIALENVTENFNMWKN LENVTENF - 19 i o 10 15 20 25 30 35 env6 ETSIKPCVKLTPLCVAMK ETSIKPCVKLSBLCITMR DQSLKPCVKLTPLCVSLK DQSLKPCVKLTPLCVTLN PCVKLTPLCV env7 NHCNTSVITESCD NTSVIT NHCNTSVIQECCD NTSVIQ TSCNTSVITQACP NTSVIT INCNTSVITQACP NTSVIT INCNTSAITQACP NTSAIT env8 YCAPPGYALLRC-NDT YCAPAGFAILKCNNKT YCAPAGFAILKCNDKK YCAPAGF^ATLKCRDKK env9 NKR PRQAWCWFKG-KWKD NERPKQAWCRFGG-NWKE N--MRQAHCNISRAKWNA D--IRRAYCTINETEWDK I--IGQAHCNISRAQWSK env 10 KGSDPEVAYMWTNCRGEFLYCNMTWFLN NCRGEFLYCN GG-DPEVTFMWTNCRGEFLYCKMNWFLN NCRGEFLYCK -GGDPEIVTHSFNCGGEFFYCNSTQLFN NCGGEFFYCN -GGDPEITTHSFNCRGEFFYCNTSKLFN ,NCRGEFFYCN -GGDPEITTHSFNCGGEFFYCNTSGLFN NCGGEFFYCN - 20 - hj •-X. "S C c 10 22 3 4 9 4 envl 1 RNYAPCHIKQIINTWHKVGRNVY CHIKQII RNYVPCHIRQIINTWHKVGKNVY CHIRQII TITLPCRIKQFINMWQEVGKAMY CRIKQFI SITLPCRIKQIINMWQKTCKAMY CRIKQII NITLQCRIKQIIKMVAGR-KAIY CRIKQII gag 1 DCKLVLKGLGTNPTLEEMLTA £ - 21 - 223 ' f V r~>£ V ■J- A * 15 20 25 % 30 35 The peptides according Co the invention may also advantageously be prepared by the standard methods applied.in the field of peptide synthesis. This synthesis may be carried out in homogeneous solution or on a solid phase. ; ' For example, use can be made of the methods of synthesis in homogeneous solution described by H0U3EWEYL in the monograph entitled "Methoden der Organischen Cheraie", edited by E. Kunsc'n, vol. 15-1 ana II, THID1E, Stuttgart, 1974. These methods of synthesis consist of condensing successive amino acids two at a tine in the required order, or of condensing amino acids with fragments already formed and already containing several amino acids in the appropriate order or, again, of condensing several fragments previously prepared in this way, it being understood that precautions will be taken to protect beforehand all of the reactive functions present on these amino acids or fragments with the exception of the amino function of one and the carboxyl function of the other which must be free to enter into the formation of the peptide bond, particularly after activation of the carboxyl function according to the well-known methods of peptide synthesis. As an alternative, recourse may be had to coupling reaction making use of standard coupling reagents of the carbodiimide type such as, for example, l-ethyl-3-(3-dimethyl~- &~inocropyl) carbocii-ice. When the amino acid used possesses an additional acid function (particular!; in the case of glutamic acid) such functions must be protected by ^"butvl ester groups, for example. In the case of step-wise synthesis in which one amino acid is added at a time, the synthesis stares preferably with the condensation of the C-terminal ariino acid with the amino acid which corresponds to the neighbouring aminoacyl *n desired sequence and it continues in this manner until the N-terminal amino acid is reached. According to another preferred method of the invention use may be made of the procedure described by R. D. MERRIFIELD in the article entitled "Solid phase peptide synthesis" (J. Am. Soc., 45_, 2149-2154). For the preparation of a peptide chain according to the procedure of MERRIFIELD it is necessary to make use of. a very porous polymeric resin to which is attached the first amino acid, the C-tej 22 3 4 9 4 i 5 10 15 20 25 30 35 the chain. This amino acid is attached to the resin through its carboxyl group and its amino function is protected by the t-butoxycarbonyl group, for example. When the, C-terminal acid is attached to the resin-as the first amino acid, the protecting group of the amine function is removed by washing the resin with an acid. In the case in which the protecting group of the amine function is the t-butoxycarbonyl group, it can be removed by treatment of the resin with trifluoroacetic acid. Subsequently, the second amino acid, which furnishes the second aminoacyl of the desired sequence counting from the C-terminal aminoacyl residue, is coupled to the deprotected amine function of the C- terminal amino acid, the first amino acid attached to the resin. The carboxyl function of this second amino acid is activated preferably for example with dicyclohexylcarbodiimide and its amine function is protected for example by means of the t-butoxycarbonyl group. In this way, the first part of the desired peptide chain is obtained consisting of two amino acids, the terminal amine function of which is protected. The amine function is deprotected as previously described and subsequently the third aminoacyl is coupled under conditions analogous to those used for the addition of the amino acid next to the C-terminal amino acid. In this way, one after the other, each of the amino acids which will constitute the peptide chain is coupled to the deprotected amine group of the portion of the peptide chain already formed and which is attached to the resin. When the desired peptide chain has been assembled in its entirety, the protecting group of the different amino acids constituting the peptide chain are removed and the peptide is cleaved from the resin by means of hydrogen fluoride, for example. The invention also relates to water-soluble oligomers of the above-mentioned peptide monomers. Oligomerization can bring about an increase of the immunogenicity of the peptide monomers according to the invention. It may be mentioned that these oligomers may, for example, contain from two to ten monomeric units without implying that this number is to be considered as limiting. 22 3 4 9 4 The monomeric units forming this oligomer are either all constituted of the polypeptide of sequence 1 or by the polypeptide of sequence 2 or by both of these polypeptides. For the preparation of the oligomer use may be made of any method of polymerization commonly used in the field of peptides, this polymerization reaction being continued until an oligomer or a polymer is obtained which contains the number of monomeric units required for the acquisition of the desired immunogenicity. One method of oligomerization or polymerization of the monomer consists in allowing the latter to react with a cross-linking agent such as glutaraldehyde. Use may also be made of other methods of oligomerization or coupling, for example that involving successive coupling of monomeric units through their terminal carboxyl and amine functions in the presence of homo- or hetero-bifunctional coupling agents. For the production of molecules containing one or more sequences of 17 amino acids such as those defined above, use may also be made of genetic engineering techniques using micro-organisms transformed by a specific nucleic acid containing the corresponding, appropriate nucleotide sequences. The invention also relates to the nucleic acids containing one or more sequences derived from the sequence of the cBNA of the virus HIV-2 ROD. These sequences, located by the numbers shown on the sequence previously described, code for some interesting peptides of the invention. Sequence coding for envl nucleotides 7850 to 7927 env2 " 8030 to 8095 env3 " 7601 to 7636 env4 " 6170 to 6247 env5 " 6294 to 6349 env6 " 6392 to 6445 env7 " 6724 to 6763 env8 " 6794 to 6838 env9 " 7112 to 7162 envIO " 7253 to 7336 envl 1 " 7358 to 7426 qaal - 1535 to 1597 - 24 - 22 3 4 9 4 C 10 15 20 25 30 35 Finally, the invention relates to nucleic acids corresponding to the SIV virus containing one or more sequences derived from the cDNA of the virus SIV-1. These sequences coding for the peptides.env 1 to env 11 and gag 1 can be located on Figure 3 by comparison with the corresponding sequences described for HIV-2. It will be obvious that the invention relates to the nucleic acid corresponding to sequences located in analogous regions of the cDNA derived from variants of HIV-2 ROD or SIV as well as all of the other nucleic acids representing modifications of those already described which result from exploiting the degeneracy of the genetic code. The invention also concerns the conjugates obtained by covaleritly coupling the peptides according to the invention (or the above-mentioned oligomers) to carrier molecules (naturally occurring or synthetic), physiologically acceptable and non-toxic, through the intermediary of complementary reactive groupings situated on the carrier molecule and the peptide. Examples of appropriate groupings are illustrated in what follows As examples of carrier molecules or-macromolectilar suppotts constituting part of the conjugates according to the invention, mention will be made of naturally occurring proteins such as tetanus anatoxin ovalbumin, serum albumins, hemocyanins, etc. ... As examples of synthetic macromolecular supports mention may be made of the polylysines or poly(D,L-alanine)-poly(L-lysine) . Other types of macromolecular supports which can be used are mentioned in the literature; usually they have a molecular weight higher than 20,000. In order to synthesize the conjugates according to the invention use may be made of known procedures such as that described by FRANTZ and ROBERTSON in Infect, and Immunity, 33, 193-198 (1981) or that described in Applied and Environmental Microbiology, (October 1981), vol. 42, No. 4, 611-614 by P. E. KAUFFMAN by using the peptide and the appropriate carrier molecule. In practice, and without implying any restriction on the use of others, it is advantageous to use the following compounds as coupling agents: glutaraldehyde, ethyl chloroformate, water-soluble carbodiimides /N-ethyl-N'(3-dimethylamino-propyl) carbodiimide, HC1/, diisocyanates,■ bis-diazobenzidine, di- and trichloro-s-triazines, cyanogen bromide as well as the coupling agents mentioned in Scand. J. Immunol., (1978), vol. 8, p. 7-23 (AVRAMEAS, TERNYNCK, GUESD0N). 22 34 9 1 5 10 15 20 25 30 35 It is possible to use any other coupling procedure which causes one or more reactive functions of the peptide, on the one hand, to react with one or more reactive functions of the support molecules, on the other. Advantageously, these reactive functions are carboxyl and amine functions which can undergo a coupling reaction in the presence of a coupling agent of the type used in the synthesis of proteins, for example, l-ethyl-3-(3-dimethylamino-propyl)-carbodiimide, N-hydroxybenzotriazole, etc ... Use may again be made of glutaraldehyde, particularly when it is required to link amino groups to each other which are situated on the peptide and the support molecule respectively. The peptides according to the invention possess antigenic properties. They may thus be used in diagnostic procedures for the detection of an infection by the HIV-2 virus. As has already been mentioned, studies have made it possible to distinguish between two groups of peptides which can be used in procedures for the detection of antibodies to the HIV-2 virus in a human biological fluid, particularly serum or cerebrospinal fluid. The first group (I) includes the gag 1 peptides. These peptides recognize anti-HIV-2 antibodies and are thus capable of detecting infection by HIV-2. To a certain extent they also recognize anti-HIV-1 antibodies. The second group (II) includes peptides which correspond more especially to those which are located in the transmembrane portion and at the terminus of the external part of the protein of the envelope. These peptides are those previously designated by env 1, env 2 and env 3. They make possible the specific recognition of the presence of antibodies to HIV-2 and thus make it possible to distinguish in a given individual past or present infections due to a HIV, more particularly between those caused by a HIV-2 and those caused by a HIV-1. The invention also relates to a composition containing at least one of the above-mentioned peptides or at least one oligomer of this peptide, characterized in that it has the capacity to be recognized by sera of human origin containing antibodies against the HIV-2 virus. The invention relates to an in vitro diagnostic procedure involving one or more peptides according to the invention for the detection of antibodies to HIV-2 in biological fluids, in particular in human sera. - 26 - 22 3 4 9 4 i 5 10 20 30 35 Generally speaking, the above in vitro diagnostic procedure comprises the following steps: - the placing in contact of this biological fluid with the said peptides, - the detection of the possible presence of an antibody-peptide complex by physical or chemical methods in the said biological fluid. In a preferred embodiment of the invention, the detection of the antibody-antigen complex is carried out by means of immunoenzymatic assays (of the ELISA type), immunofluorescence assays (of the IFA type), radioimmunological assays (of the RIA type) or radioimmunoprecipitation assays (of the RIPA type). In this sense, the invention also relates to any peptide according to the invention labelled by means of an appropriate marker of the enzymatic, fluorescence, radioactive, etc ... type. Such methods comprise for example the following steps: - deposition of defined amounts of a peptide composition according to the invention in the wells of a microtitre plate, - introduction into the said wells of increasing dilutions of the serum requiring diagnosis, - incubation of the microtitre plate, - repeated rinsing of the microtitre plate, - introduction into the wells of the microtitre plate of labelled antibodies against immunoglobulins of the blood, the antibodies having been labelled with an enzyme chosen from among those which are capable of hydrolyzing a substrate, thus leading to a change in the absorption spectrum of the latter at at least one specific wavelength, - detection of the amount of substrate hydrolyzed in comparison with a control. The invention also relates to kits for the in vitro diagnosis of the presence of antibodies to the HIV-2 viruses and, in certain cases, the HIV-1 viruses in a biological fluid. The kits contain: - a peptide composition according to the invention, - the reagent necessary for constituting a medium appropriate for performing the immunological reaction, - the reagents used to detect the antibody-antigen complex produced 1 5 10 15 20 25 30 22 3 4 9 4 by the immunological reaction. These reagents may also comprise a marker or be capable of being recognized in turn by a labelled reagent, more particularly in the case in which the above-mentioned polypeptide composition is not labelled. - a. reference biological tissue fluid not containing antibodies recognized by the above-mentioned polypeptide composition. The invention relates to the antibodies themselves formed against the peptides of the invention. It will be obvious that these antibodies are not limited to polyclonal antibodies. The invention also relates to any monoclonal antibody produced by any hybridoma capable of being formed by standard methods from the spleen cells of an animal,in particular of a mouse or a rat immunized against one of the peptides of the invention, on the one hand, and the cells of an appropriate myeloma cell line on the other, and of being selected by its capacity to produce monoclonal antibodies recognizing the peptide initially used for the immunization of the animals. The invention also relates to immunogenic compositions for the production of vaccines, the active principle of which is constituted by at least one peptide according to the invention, or an oligomer of this peptide or a peptide in the form of a conjugate with a carrier molecule, characterized in that they induce the production of antibodies against the above-mentioned peptides in amounts sufficient to inhibit the proteins of the HIV—2 retrovirus as well, and even indeed the HIV-2 retrovirus in combination with a phannaceutically acceptable vehicle. The immunogenic compositions for the production of vaccines contain more particularly in an advantageous manner at least one of the peptides previously designated as eirv4, env5, env6, env7, env8, env9, envlO, env 11 and even mixtures of them. Of the peptides suitable for constituting active principles of vaccines some are particularly preferred because they possess a basic amino acid structure corresponding to regions of the glycoproteins of the envelope which exhibit a considerable degree of conservation, not only in the HIV-2 and SIV viruses, but also in the HIV-1 viruses. These particularly preferred peptides are the peptides designated as env4, and certain peptides of env5, env6 and envlO. - 28 - 22 3 4 9 4 In a preferred embodiment of the invention the immunogenic peptides (or fragments of these peptides), suitable for constituting active principles of vaccines are chosen from among those, the formulas of which correspond to sequences which, in the glycoproteins of the envelopes of HIV-2, SIV and HIV-1, exhibit an amino acid sequence homology greater than 50%, belong to the external part of the envelope of the virus, show no or almost no deletions and contain cysteine residues favourable to the stabilization of bonds and to the constitution of bonding loops. The following peptides belong to this class of preferred peptides. env4 XVTV-YGVP-W--ATZ env5 XL-NVTE-FZ env6 XKPCVKL-PLC-Z env7 XN-S-I-Z env 10 XNC-GEF-YC-Z env 1 1 XC-I-Q-IZ Advantageous pharmaceutical compositions are constituted by solutions, suspensions or injectable liposomes containing an efficacious dose of at least one product according to the invention. These solutions, suspensions and liposomes are preferably prepared in a sterilized isotonic aqueous phase, preferably saline or a glucose solution The invention relates more particularly to those suspensions, solutions and liposomes which are suitable for administration by intradermal, intramuscular and subcutaneous injections or even by scarifications. The invention also relates to pharmaceutical compositions which can be administered by other routes, in particular by the oral route. The pharmaceutical compositions according to the invention which can be used as vaccines to stimulate the production of antibodies against the HIV-2 virus can, for example, be administered at doses varying # - 29 22 3 4 9 4 r^' a? ■> 10 15 20 25 30 35 between 10 and 500 jug/kg, and preferably between 50 and 100 pg/kg, of peptide according to the invention. These doses are cited as examples but do not imply any restriction on the dose which may be used. As has already been indicated above the various peptides which have been specified may contain modifications which do not have the effect of modifying fundamentally their immunological properties. The peptide equivalents which result form part of the claims which follow. As examples of peptide equivalents, mention will be made of those with structures corresponding to equivalent regions in the cDNAs of other variants of HIV-2, SIV or HIV-1 when these regions are aligned under conditions similar to those which were mentioned above with regard to HIV-2 ROD, SIV and HIV-1 BRU. As other examples of these peptides mention will be made of those with structures corresponding to equivalent regions in the cDNAs which were deposited with the CNCM in particular those under the numbers 1-502, 1-642 (HIV-2 IRM0), 1-643, (HIV-2 EH0) as well as, in appropriate cases, variants of HIV-1 which were deposited with the CNCM under the numbers 1-232, 1-240, 1-241, 1-550, 1-551. The peptides according to the invention can also be defined by the following formulae (in which X, Z and the dashes have the meanings indicated above): XRV-AIEKYL DQA-LN-WGCAFRQVCZ XAIEKYL-DZ X-LE-AQIQQEKNMYELQKLNSWZ XQTQQEKNZ 5 XELGDYKLVEITPIG-APT--KR Z XYKLVEITPIG-APT--KRZ 1Q X VTV-YGVP - W—AT—LFCA-Z XVTV-YGVP-W—ATZ X E--L-NVTE-F—W-NZ XL-NVTE-FZ 15 XL S-KPCVKL-PLC Z XKPCVKL-PLC-Z XS-KPCVKL-PLC-Z 2Q X N-S-I—-rC-Z XN-S-I-Z XYC-P-G-A-L-C-N-TZ 25 X A-C W— Z NKRPRQAWCWFKG-KWKD X-G-DPE NC-GEF-YC NZ 30 X C-I-Q-I G YZ 35 ,^7."' ' '• "'r ' '•'" """^Wcf^-- £>»JW: - 31 - 22 3 4 9 4 C .. ^ » V~Y 10 15 20 25 30 The invention also relates^ in addition to the peptides of SIV already described^ to the proteins coded by the cDNA of the SIV virus. It also relates to the proteins of any virus immunologically closely related to SIV-lmac, in particular any virus, the proteins and the glycoproteins of the envelope of which cross-react immunologically and the cDNAs of which exhibit a sequence homology of at least 95% and preferably at least 98%. In particular, the invention relates to: 1) the proteins and glycoproteins of the envelope encoded in the env gene and shown in Figure 3, 2) the protein GAG shown in Figure 4, 3) the protein POL shown in Figure 5, 4) the protein 0 shown in Figure 6, 5) the protein R shown in Figure 7, 6) the protein X shown in Figure 8, 7) the protein F shown in Figure 9, 8) the protein TAT shown in Figure 10. The amino acids of the previously mentioned proteins of SIV have been presented in alignment with the sequences of the amino acids of the corresponding proteins of the HIV-2 virus; the points aligned vertically between the two sequences indicate amino acids common to the proteins of the two viruses. The cDNA sequences coding for the previously mentioned proteins appear in the Figure IB. In addition to the nucleic acid sequences previously mentioned, the invention relates to any modified nucleic acid sequence which also codes for the proteins of the retrovirus SIV or one of its variants. These cDNA sequences indicated by numbers shown on the sequences previously described (Figure IB) are the following: % - 32 - 22 34 9 i 5 10 15 20 25 30 35 sequence coding for GAG. nucleotides 551 to 2068 *• POL, 1726 to 4893 <« " Q, 4826 to 5467 N X, 5298 to 5633 " i ' N R, 5637 to 5939 " M F, 8569 to 9354 n •t TAT- 1 5788 to 6084 M M ART- 1 6014 to 6130 « W TAT-2 8296 to 8 391 n M ART-2 8294 to 8548 N M ENV 6090 to 8732 Thus, the invention quite naturally relates to the proteins previously described when they are isolated from the SIV virus or when they are prepared by a method of synthesis, in particular by one of the methods already cited in connection with the synthesis of shorter peptides. The invention also relates to the use of the preceding proteins for the diagnosis of the possible presence of antibodies directed against the proteins of HIV-2, and even against the whole HIV-2 virus, and, in some cases, to their use for the purposes of diagnosis of an infection due to one of the HIV viruses. Thus, the peptide GAG encoded by the corresponding gene can be used to locate the possible presence of anti-HIV-1 or anti-HIV-2 antibodies. The env proteins are used preferably for the specific diagnosis of an infection due to HIV-2 or one of its variants, and sometimes for the diagnosis of an infection due to HIV-2 or HIV-1. Thus, the invention also relates to an in vitro diagnostic procedure for the detection of antibodies against HIV-2 and possibly against HTV-1 in biological fluids and in particular in human sera. Such procedures entailing the use of the proteins of SIV previously mentioned for diagnostic purposes have already been described in the present invention. The invention also relates to "kits" for the in vitro diagnosis of the presence of antibodies against the HIV-2 virus and, in certain cases, - 33 - 22 3 4 9 4 against HIV-1 in a biological fluid. Such kits employing the peptides previously mentioned have also been described in the present invention. The invention also relates to immunogenic compositions for the production of vaccines, the active principle of which is constituted advantageously by at least a part of the ENV protein of the SIV virus and this protein may exist as a conjugate formed with a carrier molecule. These immunogenic compositions induce the production of antibodies against the above-mentioned peptide in sufficient amounts to inhibit the proteins of the retrovirus HIV-2, and even the retrovirus HIV-2 itself. However, the use for diagnostic purposes of the proteins of SIV is in no way limited only to the proteins ENV and GAG. Other proteins among those described may be considered for the preparation of compositions for diagnosis and even as vaccines. </div>

Claims

<div id="claims" class="application article clearfix printTableText"> - 34 - O r ' r- /» 1 WHAT WE CLAIM ISt-

1) A peptide having immunological properties in common with chose of the peptide backbone of the glycoprotein of the envelope of the viruses of the HIV-2 class, characterized in that it also has a peptide structure in common with the peptide backbone of the glycoprotein of SIV-1.

2) A peptide having immunological properties in common with those of the peptide backbone of the glycoprotein of the envelope of the viruses of the HIV-2 class, these peptides containing a number of amino acid residues not exceeding 40, characterized in that it also has a peptide .structure in common with the peptide backbone of the glycoprotein of SIV-1.

3) A pqatide according to Claim 2 characterized by one of the formulae: XRV-AIEKYL-DQA-LN-WGCAFRQVCZ or XAIEXYL-DZ in which X and Z are respectively NHj and OH groups when the N-terminal amino-acid group and the C-terminal amino-acid group are respectively devoid of these and OH groups or, in cases in which the immunological properties of the peptide not containing these groups are found to be not substantially modified, they may represent groups containing from 1 to 5 amino acid residues and each of the dashes correspondsto an aminoacyl residue chosen from among those which enable the peptide defined above to conserve the immunological properties of one of the following sequences: AIEKYLQDQ , RVSAIEXYLKDQAQLNAWGCAFRQVC or AIEKYLKDQ .

4) A peptide according to Claim 2 characterized by one of the formulae: X - LE - AQIQQ EXNMYELQKLN S W Z or XQIQQEXNZ in which X and Z are respectively NH2 OH groups when the N-terminal amino-acid group and the C-terminal amino-acid group are respectively devoid of these SHj and OH groups or, in cases in which the immunological properties of the peptide not containing these groups are found to be not substantially modified, they may represent groups f—— containing from 1 to 5 amino acid residues and each of the dashes corresponds to an aminoacyl residue chosen from among those which enable the peptide defined above to conserve the immunological properties of one of the following secuences: [,'<r i" 13 AUGtf SLEQAQIQQEKNMYELQKLNSW' QIQQEKN or LLEEAQIQQEKNMYELQKLNSW .

5) A peptide according to Claim 2 characterized by one of the formulae XELGDYKLVEITPIG-APT--KR Z XYKLVEITPIG-APT—KRZ in which X and Z are respectively NH, and OH groups when the N-terninal amino-acid group and the C-terminal amino-acid group are respectively devoid of these NHj and OH groups or, in cases in which the immunological properties of the peptide not containing these groups are found to be not substantially modified, they may represent groups containing from 1 to 5 amino acid residues and each of the dashes correspondsto an aminoacyl residue chosen from among those which enable the peptide defined above to conserve the immunological properties of one of the following sequences: ELGDYKLVEITPIGFAPTKEKRYSSAH , YKLVEITPIGFAPTKEK , ELGDYKLVEITPIGLAPTNVKRYTTG or YXLVEITPIGLAPTNVK .

6) A peptide according to Claim 2 characterized by one of the formulae: X VTV-YGVP-W—AT—LFCA-Z or XVTV-YGVP-W--ATZ in which X and Z are respectively US2 OH groups when the N-terminal amino-acid group and the C-terminal amino-acid group are respectively devoid of these NHj and OH groups or, in cases in which the immunological properties of the peptide not containing these groups are found to be not substantially modified, they may represent groups containing from 1 to 5 amino acid residues and each of the dashes corresponds to an aminoacyl residue chosen from among those which enable the peptide defined above to conserve the immunological properties of one of the following sequences: VTYFYGVPTWKNAT , CIQYVTVFYGVPAWRNATIPLFCAT / VTVFYGVPAWRNAT, EKLWVTVYYGVPVWKEATTTLFCAS or VTVYYGVPVWKEAT. - 36 - O <"> r> A r- j dc,Cjf\;; 4 '/ © c iO ry 1Z> c 25 30 7) A peptide according to Claim 6 characterized by one of the formulae: VTVFYGVPTWKNAT , CIQYVTVFYGVPAWRNATIPLFCAT , VTVFYGVPAWRNAT, EKLWVTVYYGVPVWKEATTTLFCAS , VTVYYGVPVWKEAT / EDLWVTVYYGVPVWKEATTTLFCAS , VTVYYGVPVWKEAT* DNLWVTVYYGVPVWKEATTTLFCASOr VTVYYGVPVWKEAT- 8) A p^stideaccording to Claim 2 characterized by one of the formulae: x E--L-NVTE-F—W-NZ or XL-NVTE-FZ in which X and Z are respectively NH2 and OH groups when the N-terminal amino-acid group and the C-terminal amino-acid gxoup are respectively devoid of these NHj and OH groups or, in cases in which the immunological properties of the peptide not containing these groups are found to be not substantially modified, they may represent groups containing from 1 to 5 amino acid residues and each of the dashes corresponds to an aminoacyl residue chosen from among those which enable the peptide defined above to conserve the immunological properties of one of the following sequences: L-NVTE , DDYSELAL-NVTESFDAWEN , PNPQEVVLVNVTENFNMWKN or LVNVTE . 9) A peptide according to Claim 8 characterized by one of the formulae: DDYQEITL-NVTEAFDAWNN / L-NVTEAF / DDYSELAL-NVTESFDAWEN , L-NVTESF , PNPQEVVLVNVTENFNMWKN » LVNVTENF / PNPQEIELENVTEGFNMWKN» -S3PSSC. ■ - 37 - 223494 LENVTECF, PNPQEIALENVTENFNMWKN or LENVTENF- 10) A peptide according to Claim 2 characterized by one of the formulae: XL S-KPCVKL-PLC Z , XXPCVKLTPLCVZ car XS-KPCVKLTPLCVZ- in which X and Z are respectively NH2 and OH groups when the X-teraina.1 ami no—acid group and the C-termirial amiao-acid group are respectively devoid of these SHj and OH groups or, in cases in which the asunuaological properties of the peptide not containing these groups are found to be not substantially modified, they may represent grouss containing from 1 to 5 amino acid residues and each of the dashes corresponds to an aminoacyl residue chosen from among those which enable the peptide defined above to conserve the immunological properties of one of she following sequences: LFETSIKPCVKLTPLCVAMK, LFETSIKPCVKLSPLCITMR, LWDQSLKPCVKLTPLCVSLK, KPCVXLTPLCV, KPCVKLSPLCI 'or SLKPCVKLTPLCV. 11) A peptide according to Claim 10 characterized b\^ne~Of--the-£^lowing structures: LFETSIKPCVKLTPLCVAMK # LFETSIKPCVKLSPLCITMR/ LWDQSLKPCVKLTPLCVSLK » LWDQSLKPCVKLTPLCVTLN/ PCVXLTPLCV OC KPCVKLSPLCI• 12) A peptide according to Claim 2 characterized in that it contains basic structure: X N-S-I C-Z or XN-S-I-Z

7-5;- \. • . -38" 22349 x o 3r * yi O 25 in which X and 2 are respectively SHj 4116 08 5-oups when the N-terminal amino-acid group and the C-terminal amino-acid group are respectively devoid of these MH2 and OH groups or, in cases in which the immunological properties of the peptide not containing these croups are found to be not substantially modified, they may represent grouos containing from 1 to 5 amino acid residues and each of the dashes correspondsto am aminoacyl residue chosen from among those which enable the peptide defined above to conserve the immunological properties of one of the following sequences: NHCNTSVITESCD, NTSVIT, NHCNTSVIQECCD, NTSVIQ, TSCNTSVITQACP or NTSVIT. 13) A peptide according to Claim 12 characterized by one of the following formulae: NHCNTSVITESCD f NTSVIT, NHCNTSVIQECCD , NTSVIQ f TSCNTSVITQACP NTSVIT t INCNTSVITQACP ntsvit / incntsaitqacp or ntsait- 14) a peptide according to Claim 2 characterized by the following formula: XYC-P-G-A-L-C-N-TZ in which X and 3 are respectively XH-, and OH groups when the N-terminal amino-acid group and the C-terminal amino-acid group are respectively devoid of these MH-, and OH groups or, in cases in which the immunological properties of the peptide not containing these groups are found to be not substantially modified, they may represent grouag containing from 1 to 5 amino acid residues and each of the da§ corresponds to an aminoacyl residue chosen from among those o /»* -i .-i-i 4 ' 2S 3D -f the peptide defined above to conserve the immunological properties of one of the following- sequences: YCAPPGYALLRC-NDT or YCAPAGFAILKCNNKT. 15) A peptide according to claim 2 or to claijn 14 diaracterized by one of the formulae: YCAPPGYALLRC-NDT / YCAPAGFAILKCNNKT / YCAPAGFAILKCNDKK qj. YCAPAGFAILKCRDKK „ 16) A peptide according co Claim 2 characterized by the formula: x A-C W—Z in which X and Z are respectively NH2 and OH groups when the -N*-terminal amino-acid group and the C-terminal amino-acid group are respectively devoid of these SH^ and OH groups or, in cases in which the immunological properties of the peptide not containing these groups are found to be not substantially modified, they may represent groups containing from 1 to 5 amino acid residues and each of the dashes corresponds to an aminoacyl residue chosen from among those which enable the peptide defined above to conserve the immunological properties of one of the following sequences: NKRPRQAWCWrKG-KWKD , NERPKQAWCRFGG-NWKE ©r N--MRQAHCNISRAKWNA . 17) A peptide according to Claim 16 characterized by the following formulae: NKRPRQAWCVIFKG-KWKD / NERPKQAJWCRFGG-KWKE , N--MRQAHCNISRAKWNA ' D--IRRAYCTINETEWDK or I--IGQAHCNISRAQWSK . 18) A peptide according to Claim 2 characterized by the following formulae: X-G-DPE NC-GEF-YC NZ or XNC-GEF-YC-Z in which X and 3 are respectively SHj and OB croups when die amino-acid grouc and the C-cerainal amino-acid group are regjje " A'A „* \ - 40 - 223494 10 15 20 25 30 35 devoid of these UHj and OB groups or, in cases in which the inmuno logical properties of the peptide not containing these groups are found to be not substantially modified, they may represent groups containing from 1 to 5 amino acid residues and each of the dashes corresponds to an aminoacyl residue chosen from among those which enable the peptide defined above to conserve the immunological properties of one of the following sequences: NCRGEFLYCN GG-DPEVTFMWTNCRGEFLYCKKNWFLN NCRGEFLYCK , -GGDPEIVTHSFNCGGEFFYCNSTQLFN NCGGEFFYCN . or 19) A peptide according to Claim 18 characterized by one of the following structures: NCRGEFLYCN , GG-DPEVTFMWTNCRGEFLYCKMNWFLN , NCRGEFLYCK , -GGDPEIVTHSFNCGGEFFYCNSTQLFN, NCGGEFFYCN , -GGDPEITTHSFNCRGEFFYCNTSKLFN, NCRGEFFYCN, -GGDPEITTHSFNCGGEFFYCNTSGLFN or NCGGEFFYCN . 20) A peptide according to Claim 2 characterized by one of the following formulae: --C-I-Q-I G---YZ XC-I-Q-IZ X- or in which X and Z are respectively NH2 0B 9rouPs when the N-terminal amino-acid group and the C—terminal amino-acid group are respectively devoid of these NH^ and OH groups or, in cases in which the immunological properties of the peptide not containing these groups are found to be not substantially modified, they may represent groups containing from 1 to 5 amino acid residues and each of the dashes corresponds to an aminoacyl residue chosen from among those which enable the oeptide defined above to conserve the immunological properties of one of the following secuences: RNYAPCHIKQIINTWHKVGRNVY , CHIKQII / RNYVPCHIRQIINTWHKVGKNVY, CHIRQII , TITLPCRIKQFINKWQEVGKAMY car CRIKQFI. 21) A peptide according to Claim 20 characterized by one of the following structures: CHIKQII / RNYVPCHIRQIINTWKKVGKNVY, CHIRQII , TITLPCRIKQFINKWQEVGKAMY, CRIKQFI , SITLPCRIKQIINMWQKTCKAMY , CRIKQII , NITLQCRIKQIIKMVAGR-KAIY or CRIKQII . 22)An antigenicpeptide gagl, characterized by one of the basic structures: XDCKLVLKGLGMNPTLEEMLTAZ or XDCKLVLKGLGTNPTLEEMLTAZ in which X and 2 are respectively NH2 and OH groups when the S-terminal amino-acid group and Che C-teraiaal amino-acid group axe respectively devoid of these NH2 and OH groups or, in cases in which the immunological properties of the peptide not containing these groups are found to be not substantially modified, they say represent groups containing from 1 to 5 amino acid residues and each of the dashes corresponds to an aminoacyl residue chosen from among those which en. the peptide defined above to conserve the immunological properties of one of the following sequences: DCKLVLKGLGTNPTLEEMLTA 23) A DNA molecule having a nucleotide sequence of at least 18 nucleotides characterized in that it contains all or part-of the sequence of the nucleic acids defined in Figure IB. - 1,2 - O 7 494 £ 'A' £ s % s 6 c 10 15 20 25 30 35 24) A DNA molecule having a nucleotide sequence of at least 18 nucleotides characterized in that it contains all or part: of the sequence of the nucleic acids defined in Figure 1C. 25) A DNA molecule having a nucleotide sequence according to claim 23, characterized in that it comprises the nucleotide sequence: GAG extending from POL Q X R F TAT- 1 ART-1 TAT-2 ART-2 LTR ENV nucleotides 550 to 2068 , . 1726 to 4893 " - 4826 to 5467 ' 5298 to 5633 ' 5637 to 5939 ' 8569 to 9354 ' 5788 to 6084' 6014 to 6130' 8296 to 8391 ' 8294 to 8548/ 8950 to 9468 end 1 to 316 or 6090 to 8732 • 26) A peptide of at least 6 amino acids having a peptide structure in common with the peptide backbone of SIV-1, characterized in that it comprises all or part of the following amino acid sequences.: ENV shown in figure 3 / 4 , 5 , 6 #■ 7 , 8 / 9 r 10 or 11 • 27) Recombinant nucleic acid characterized in that' it comprises" t-he totality or part of a cDNA according to any one of the Claims 23 to 25 inserted in a nucleic acid derived from a vector. 28) Recombinant nucleic acid according to Claim 27, characterized in that it is labelled. POL Q R X F TAT ART -rg&r- T' 223494 29)An antigeniccomposition containing the gag peptide according to Claim 26 , at least one gagl peptide according to Claim 22 or at least an oligomer of this peptide, characterized in that the composition has the capacity to be recognized by biological fluids of human origin, in particular by sera containing anti-HIV-2 antibodies and, to a certain extent, anti-HIV-1 antibodies. 30)An antigeniccomposition containing the env peptide according to Claim 26 or at least a peptide according to Claims 3, 4 and 5 or at least an oligomer of this peptide, characterized in that the composition specifically recognizes the presence of antibodies against HIV-2. 31)An immunogeniccomposition containing all or part of the env peptide according to Claim 26 or at least a peptide or at least an oligomer of this peptide or this peptide in the form of a conjugate with a carrier molecule according to Claims 6 to 21 in combination with a pharmaceutical^ acceptable vehicle for the production of vaccines, characterized in that the composition induces the production of antibodies against the above-mentioned peptides in sufficient amounts for them to be able to inhibit in an efficacious manner the proteins of the retrovirus HIV-2, or even the entire HIV-2 retrovirus. 32)An immunogeniccomposition according to Claim 31 characterized in that it contains peptides, the formulae of which correspond to sequences in the glycoproteins of the envelope of HIV-2, SIV-1 and HIV-1 which exhibit an amino acid sequence homology greater than 50Z. 33)An immunogeniccomposition according to one of the Claims 31 or 32, characterized in that it contains at least one peptide or at least one oligomer of this peptide or this peptide in the form of a conjugate with a carrier molecule chosen from among env£, env5, env6 and envlO. 3&)An in vitro diagnostic procedure for an infection by HIV-2 in a biological fluid comprising: - the placing in contact of this biological fluid with at least one peptide according to one of the Claims 1, 2, 3, 4, ^,^22 or a conjugate of these peptides with a carrier molecule, or with the peptiBfes^ env according to Claim 26 I >o may, 99,s] . ! v 7 - 44 - 22349 - the detection of the possible presence of an antibody-antigen complex by physical or chemical methods in the said biological fluid. 35)An in vitrodiagnostic procedure for an infection by HIV-2 in a biological fluid according to Claim 34, characterized in that the antibody antigen complex which may have formed is detected by means of inimuno-enzymatic assays (of the ELISA type), immunofluorescence assay (of the IFA type), radioimmunological assays (of the RIA type) or radioinmuno-precipitation assays (of the RIPA type) . 36) A kit for che in vitro diagnosis of an infection by HIV-2 in a biological fluid characterized in that it comprises: - a peptide composition containing a peptide according to one of the Clsic. 1 to 5, 22 or a mixture of these peptides or a conjugate of these peptides with a carrier molecule, or the peptides gag or env according to Clair 26, - a reagent for constituting an appropriate medium for the performance of an immunological reaction, - one or more reagents, possibly labelled, for the detection of the antibody-antiger. complex formed as a result of the immunological reaction. - a reference biological fluid not containing antibodies recognized by the above-mentioned peptide composition. 37) A peptide as claimed in claim lf substantially as herein described with reference to any one of the examples. 38) An antigenic peptide as claimed in claim 22, substantially as herein described with reference to any one of the examples. 39) A DNA molecule having a nucleotide sequence as claimed in claim 23, substantially as herein described with reference to any ' one of the examples. 40) Recombinant nucleic acid as claimed in claim .27 substantially as herein described with reference to any one of the examples. 41) An antigenic composition as claimed in claim 30* substantially as herein described with reference to any one of the examples. 42) An immunogenic composition as claimed in claim 30 substantially as herein described. _ 43) An in vitro diagnostic procedure as claimed in claim 34 substantially as herein described. BALDWIN SON & CAREY </div>