SE468181B - HIV-1 gp41 peptide, vaccine and antigen containing the peptide and also diagnostic test using the peptide - Google Patents

Abstract

A synthetic antigen in the form of a peptide, the sequence of which corresponds to parts of the proteins of the HIV-1 virus and which peptide has the formula: Asp-Gln-Gln-Leu-Leu-Ile-Trp-Gly-Cys-Ser-Gly-Lys-Leu-Ile- Cys-Thr-Thr-Ala-Val-Pro-Trp-Asn-Cys-OH. The peptide is immunoreactive with HIV-1-specific antibodies and intended to be used for diagnosis of HIV-1 and also to give rise to production of antibodies to HIV- 1 in animals and humans. <IMAGE>

Description

468,181 g of patient serum inhibit the reaction. But despite this, Wellcome's tests can occasionally give False positive reactions.

Other alternative methods of obtaining reagents that provide specific and safe answers to HIV-1 tests are based on molecular biological methods. The HIV-1 virus gene has been cloned and the entire nucleotide sequence is known (Muesing M.A. Douglas H.

Smith D. Cabradilla, Charles V.Benton, Laurence A. Lasky and Daniel J. Capon. Nucleic acid structure and expression of the human AIDS / lymphadenopathy retrovirus. Nature 313: 450-458, 1985.) (Sanchez-Pescador; Ray. Michael D. Power. Philip J.

Barr, Kathelyn S. Steimer. Michelle M. Stemoien, Sheryl L.

Brown-Shimer. Wendy W. Gee. Andre Renard. Anne Randolph, Jay A. Levy, Dino Dina, Paul A. Luciw. Nucleotide sequence and expression of an AIDS-associated retrovirus (ARV-21. Science 227: 484-492, 1985.) It is known which parts of the HIV-1 genome encode the different viral proteins. Interesting parts of the HIV-1 genome have been cloned into bacteria and the bacteria have been made to synthesize immunologically active HIV-1 proteins that can be used in diagnostic tests. An example is that part of the envelope protein gp41 of the HIV-1 virus has been synthesized in bacteria and used for serological diagnosis of AIDS (Cabradilla Cirilo D. Jerome E. Gropman. Joan Lanigan, Mark Renz, Laurence A. Lasky and Daniel J. Capon Seodiagnosis of antibodies to the human AIDS retrovirus with a bacterially synthesized ENV polypeptide. Biotechnology 4: 128-133, 1986.) A test built around a synthetic bacterial antigen is considered specific, sensitive and does not give any false positive reactions. But in general, there is always a disadvantage with bacterially produced proteins and that is the high degree of purification required to remove all bacterial proteins from the antigen. As has been said for cell-grown virus, intensive purification processes can damage the virus proteins so that they lose their antigenic properties to some extent.

The regions of a protein that act as antigen in serological tests e.g. ELISA is called epitopes. the number of epitopes in an ordinary protein amounts to about 5-10 different »gp 3,468,181 epitopes. Each epitope region is made up of 6-7 different amino acids that can either lie in a sequence and form a continuous epitope or. which is more common. radiates several parts of the amino acid chain into an area that forms a discontinuous epitope (Novotny Jiri, Mark Handschumacher and Robert E. Bruccoleri Protein antigenicity: A static surface property Immunology today 8: 25-31, 1987). Modern developments in the field of peptide synthesis have made it possible to synthesize peptides which may contain up to about 100 amino acids. With peptide synthesis methodology, it should therefore be possible to extract the so-called continuous epitopes from HIV-1 virus structural proteins. It is then important to first select the HIV-1 virus proteins that are particularly interesting as antigen in an ELISA test. Our own and others' assessments show that gp41 is such an interesting protein.

The nucleotide sequence of gp41 is known and the nucleotide sequence can easily translate into the corresponding amino acid sequence. From this long amino acid sequence, it is important for synthesis to select areas that can correspond to diagnostically interesting continuous epitopes.

In a work by Wang and co-workers (Wang James JG. Samuel Steel, Rosanne Wiesniewolski and Chang Yi Wang Direction of antibodies to human T-lymphotropic virus type III by using a synthetic peptide of 21 amino acid residues corresponding to a highly antigenic segment of gp41 envelope protein. Prod Natl Acad Sci USA 83: 6159-6183, 1986) presents such a synthetic peptide. The peptide in question consists of 21 amino acids with the following sequence. Arg-Ile-Leu * Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys-Ser, and corresponds to amino acids Nos. 586-606 of the precursor of the envelope protein to HIV-1 (HTLV III). A combined serological. chemical analysis of this peptide and adjacent peptides showed how important certain amino acids were for serological reactivity. Of all the different combinations, removal of the four carboxy-terminal amino acids Trp-Gly-Ser gave only a small change. reduction, of the serological reactivity. In contrast, some amino acids in the amino-terminal part of the peptide were of great importance for the 468 181 q serological reactivity, in particular Arg-1, Ile-2 and Lys-10. Wang and colleagues have used this peptide in ELISA to measure antibodies to HIV-1 / HTLV III with good results. The peptide does not give any false positive reactions, and only a few false negative reactions.

In U.S. Pat. U.S. Patent 4,529,783 discloses several immunologically reactive synthetic peptides corresponding to portions of the HIV-1 virus proteins for the diagnosis of AIDS and AIDS-related diseases. Of particular interest is peptide V (39) having the sequence Arg-I1e-Leu-Ala-Val-Glu-Arg-Tyr-Leu-Lys-Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys -Ser-Gly-Lys-Leu-Ile-Cys-X, where X is OH or NH 2. which corresponds to a portion of gp41 encoded by base pairs (bp) 7516-7593 of the HIV-1 genome. Peptide V (39) responded with 23/24 (95.8Z) serum samples from patients with confirmed HIV-1 infection.

For other viruses, synthetic peptides have been used as vaccines. (Van Regenmortel MHV Synthetic peptides as viral vaccines ann Inst Pasteur / Virol 137E, 497-528. 1986 and it may also be possible to prepare HIV-1 vaccines based on synthetic peptides, a first step in this direction has been presented by Kennedy.Esssex and co-workers (Kennedy RC.Henkel, D.Pauletti, JS allan, TH Lee M. Essex, GR

Dreesman Antiserum to a synthetic peptide recognizes the HTLV III envelope glycoprotein Science 23fl4% 556-1559, 1986). They have synthesized a peptide corresponding to amino acids 735-752 in gp16O a envelope protein in HIV-1. This peptide reacts with human antisera containing antibodies directed against HIV-1.

The authors have then concluded that the peptide is care to try as a vaccine against HIV-1 and also as a possible diagnostic antigen. The object of the present invention is to synthesize an immunoreactive peptide to be used for the diagnosis of HIV-1 partly for the determination of antibodies and also for the immunization of animals to develop antibody reagents which can be used for the determination and search for HIV-1 antigen. in patient samples during the early phase of HIV-1 infection when patients have not yet developed antibodies. An antigenically active peptide should also be able to be tested for efficacy as a vaccine against HIV-1.

Among the large amount of synthetic peptides we have prepared and tested, we have found a peptide that exhibits particularly good properties as a diagnostic reagent in ELISA for detecting antibodies to HIV-1 in patient sera. This peptide corresponds to an amino acid sequence in gp41 and the peptide has been named "A5" by us.

The amino acid sequence of A5 is as follows: Asp-Gln-Gln-Leu-Leu-Gly-Ile-Trp-Gly-Cys-Ser-Gly-Lys-Leu-Ile-Cys-Thr-Thr-Ala-Val-Pro-Trp- Asn-Cys-OH.

Using the same system for numbering Wang (7), the peptide of the invention corresponds to amino acid no. 598-618. The peptide that Wang has synthesized corresponds to amino acids 586-606.

In the inventive peptide A5, an amino acid Cys has been added at position 519 in the thinkboxy terminal part. In the real peptide is the amino acid Ala. We have found it essential to give the peptide this artificial supplement in order for it to express beneficial properties when used as a diagnostic reagent in ELISA. The large difference with the cheek peptide should be noted. Nang tested a large number of peptides in the region 586-606 but it was only in the amino terminal part around 586-596 that Wang found amino acids critical for reactivity. In our synthesis, compared to Wang, we have gone far to the right in an area that Nang found uninteresting and have here defined a peptide with extremely good properties as a diagnostic reagent.

In particular, it has been subjected to extensive testing against large series of known positive and negative sera. As can be seen from the accounts below, the result has been excellent. 468 181 Description of the drawings The invention will be described in more detail below in the form of exemplary embodiments and test results with reference to the accompanying drawing figure. which is a histogram showing ELISA values obtained using the peptide gp41A5 of the invention as antigen.

Synthesis of peptide gg4] A5 Peptide gp41A5 has been synthesized according to Merrifield's stepwise solid-phase strategy using an automatic peptide synthesis apparatus - Applied Biosystem 430. The constituent amino acids (Nova Biochem, Switzerland) used in the synthesis have been protected in a conventional manner in amino terminals more tertylbutylox (t-BOC) groups and where stable compounds have been used to protect the side chains of the amino acids. t-BOC amino acids have been sequentially linked p- a resin-p-methylbenzylhydrylamine-RMB-2100 (Peptides International, Luisville, USA).

Extraction and cleavage of peptide gp41A5 has been performed via conventional methodology with a dedicated apparatus setup (Applied Biosystem). The cleavage was performed in the presence of hydrogen fluoride, dimethyl sulfoxide and anisole in the ratio 10: 1: 1 per part of peptide bound to resin.

Final peptide product has been analyzed by HPLC reversed phase (Waters). n me- 'ke' ns en ss "SA et 'k fö v' v 'r ar oe' 4 A5 Conventional ELISA protocol is used according to ïöljandez - Microtiter plates coupled with peptide according to previously adsorb human serum in dilution 1/100 (dilution liquid: PBS _ 0 _ containing 1 Z BSA) for 90 minutes at 37 C in a humid chamber.

In 468 181 - Serum is poured off and the plates are washed 3 times with PBS containing 0.052 Tween 20 (PBS-Tween).

Adsorption with antihuman igG conjugate alkaline phosphatase.

(Dakopatt's D-338 predicted in PBS 1: 800) - Incubate for 90 minutes at 37UC in a humid chamber.

- Wash 3 times with PBS-Tween.

Addition of alkaline phosphatase substrate (Sigma 104) 1 mg / ml, 200 μl / well. - 30-60 minutes conversion of substrate and reading with multiscan multichannel spectrophotometer.

A '' t tter a t Microtiter plates intended for antibody analysis of patient serum by enzyme-linked immunosorbent assay (ELISA) have been coated with peptide according to the following protocol. - 96-hole microtiter plates (Greiner alt. Nunc) Fill with 50 μl poly-L-lysine (Sigma P 1399) at a concentration of 1.0 mg / 100 ml. phosphate buffer (PBS pH 7.2) and adsorbs at 20 ° C for 1 minute.

- The plates are poured and washed with 200 ul PBS / hole. 50 μl of peptide gp41A5 is added to each hole in the microtiter plate. At a concentration of 10 pg / ml predicted in PBS. - 50 μl glutaraldehyde (G5882 Sigma) predicted to 0.5 Z in PBS _ _. . The peptide solution is added to the microtiter plate. Far stand 20 C for 15 minutes.

- The solutions are poured off and the plates are washed twice in PBS. 468 181 s - 200 μl 100 mM glycine (diluted in H 2 O containing 0.11 bovine serum albumin BSA) is added to the wells and allowed to react for 30 minutes at 20 ° C.

- The solutions are poured off and the plates are washed twice in PBS.

The plates must be drained and stored at -20ÛC for use. i humana sera.

Peptide gp41A5 has been assayed for concentration used in ELISA and found to be immunoreactive when bound to poly-L-lysine in microtiter plates at a concentration of 500 pg / ml - 0.1 μl / ml. The optimal concentration of gp41A5 has been shown to be 10 ug / ml and has been used in all further documentation.

Screening of 101 documented positive sera and 170 negative sera is illustrated in Figure 1.

Positive sera consisted of high and low positives from Tel Aviv in Israel, Gothenburg, Sloan-Kettering Ca Ce New York (3 and 4, +40 +54).

Negative sera originated from transplant recipients. leukemia patients, false positive sera from other ELISA tests and blood donors (25 + 21 + 28 + 105). These negative sera show a very low background = 0.100 OD units. Limit value ¥ to be judged as a positive reaction has been set to the background (M = 0.100) + 3 standard deviation units (0.092) = 0.100 + 3 X 0.092 = 0.367.

With these criteria for positivity, 4 separate tests have given clearly positive results for 101 sera, which have previously been shown to be positive with other methodologies.

Mass screening of 413 documented positives (with Wester blot 01 fl 9 confirmation test) shows according to Table 1 positive with gp41A5. 468,181 that all were 1024 blood donors, all were shown to be negative. A further 182 sera from patients with infectious diseases or malignancies who frequently give false positive antibody results using conventional methodology showed negative against gp41A5. 82 patient sera selected reactions to conventional ELISA HIV-1 all give negative reactions to gp41A5. 40 intermediate with Du positive sera (positive / negative?) With commercially all antigens test showed, Vara on the basis of false positives gave borderline activity Pont with gp41A5 a result that completely coincided (100ZT with high-sensitivity confirmation test western blot.

Table 1 Implementation of gn 41-A Deptide ELISA True positive sera 413 sera positive with other ELISA 413 positive with gp 41 A: ie. 413/413 (1001) True negative sera 1,024 blood donor sera: "Betrayal" true negative sera: Wasserman reaction (WR) positive sera: Treponema Palladium positive (WR) Anti-complement reactive sera: Epstein-Barr virus IgM-positive sera: Graft recipient sera: (Western blot positive): -tests: (Western blot negative): Sêraå OOOCDO positive positive positive positive positive positive with with with with with with with gp41A5 gp41A5 gp41A5 gp41A5 gp41A5 gpk1A5 468 181/0 21 Leukemia patient with goa sera: 0 positive 0 / 1.206 (OZ) False positive sera in other ELISA tests: 14 false positive in Abbott-ELISA D positive with gp41A 38 false positive in Behring-ELISA D positive with gp41A 8 false positive in Dupont-ELISA 0 positive with gp41A 4 false positive in EN1 ELISA D s''ve 41 D / 52 (OZ) gp41A5 peptide ELISA was performed with 40 sera that gave "grâzons" reactions (below the limit but above the negative control values) in tests with Dupont tests; which resulted in: 8 Western blot positive sera: 3 2 western blot negative sera: As a comparison with the synthesized peptide V (39) according to the Peptide, in ELISA test 8 positive with gp41A5 32 negative with gp41A5 (2 gave "gray zone" reactions) the peptide according to the invention U.S. U.S. Patent 4,829,783. and gave 221/233 (94.8Z) positive reactions with known HIV-1 positive serum samples. Twelve false negative peptide V (39) reactions a false positive in 1 sera. negative Serum samples negative by western blot analysis with HIV-1 false positive and negative Western blot analysis.

It should be noted peptide SMZB4 according to Wang et al. 83: 6159-6163. And peptide V (39) The inventive peptide amino acid frequency of the known peptides plus an additional sequence region of gp41 SM284 and V (39). Wang et al. reactions were determined (Proc. according to gp41A5 thus. 102 In addition (0.98Z) SOm gave of confirmed and All positive proteins. were confirmed by gp41A5 having a partial overlap with Natl. Acad. (1986) patent 4,629,783.

Sci.

U.S. thus containing the carboxy-terminal part of the two corresponding one at the carboxy-terminal side of the peptides, however, considered that the carboxy-terminal part of peptide SM284 was immunologically less important compared to the amino-terminal part of peptide SM284.

However, according to the invention, it has been found. that by extending the permutation of gp41A5 to the carboxy-terminal portion of gpki, a peptide having excellent antigenic properties for detecting AIDS-specific antibodies in human sera has been obtained. This peptide shows both a higher reactivity and greater specificity in ELISA and western blot than the two peptides V (39) and SM284.

Table 2 below shows that gp41A5 reigns with all 233 (1002) established positive HIV-1 sera. These results were obtained from the same 233 sera tested with peptide V (39), which gave 12 false negative reactions. The mean limit value for a positive reaction in the ELISA tests was an O.D.4Ü5 of about 0.3.

Any reading below 0.3 was considered a negative reaction. It is clear that the serum samples 7.19.4D.44,48.6D.101.482,511, 324.375 and 393 showed no reactivity with peptide V (39) but superior reactivity with gp41A5.

Table 3 shows a further comparison of the immunological reactivity of gp41A5 with peptide V (39). Thirty-eight of the 233 serum samples presented in Table 2 were identified as positive by western blot analysis. These positive sera plus eight determined negative serum samples (Western blot) were regulated with the peptides gp41A5 and V (391: ifallallell ELISA tests with positive reactions, all with a D.D.¿s exceeding 0.3.

In addition, a non-HIV-1 antigen (negative antigen) was used as a control. It is striking that gp41A5 did not show any false negative or false positive reactions. Thus, the inventive peptide gp41A5 was found to surprisingly and unexpectedly give ELISA results that are clearly 1002 correct. 400 101 M TABLE 2 Immunological reactivity determined by ELISA.between HIV-1 antibodies 1 233 established positive serum samples and synthetic peptide antigens corresponding parts of gp41 Antigen Se * ““ Pr ° V nr- v (39) gp41As 10 4 0.436 1.4009 7 0.13 12 0.330 2.222 13 0.030 2.090 17 0.005 1.527 15 10 1.336 2.711 19 0.209 2.922 23 1.706 2.022 24 0.405, 2.010 25 0.404 2.635 20 26 1.443 2.000 27 0.010 2.029 29 1.399 2.459 30 1.009 2.500 31 0.737 2.343 25 32 1.244 2.902 33 1.332 2.071 35 0.930 2.714 37 1.007 2.059 - 30 1.020 "2.370 30 39 1.362 2.529 40 0.227 1.495 41 1.202 2.692 42 0.731 2.209 43 0.522 1.970 35 44 1.199 2.443 45 1.096 1.044 46 1.456 2.796 47 0.005 2.036 40 0.195 2.592 40 49 1.532 2.009 51 0.712 * 2.579 53 1,217 2,714 54 0.002 2,423 55 1,444 2,567 45 56 0.032 2,511 57 1,104 2,400 50 1,510 2,232 59 1,919 2,726 00 0.219 1,399 50 70 0.409 1,020 79 0.927 2,247 101 0.205 0.003 402 0.120 1.310 511 0.240 1.030 55 130 0.340 1,741 134 1,256 2,425 137 1,095 2,413 140 0.952 2,364 1 0 15 2o_ 25 30 35 40 45 50 TABLE 2 (continued) A3 468 181 S Anticen erumprov nr. V (39) gp4lA5 225 1,464 2,548 232 1,877 2,654 243 0.644 2,397 272 1,968 2,767 273 1,404 2,660 274 2,037 2,821 275 1,786 2,609 276 1,900 2,745 277 1,106 2,243 278 1,203 2,354 279 0.555 2,579 285 1,417 2,897 2,870 2,897 2,897 2,897 2,897 2,897 2,897 2,897 2,897 2,897 2,287 2,097 2,897 2,897 2,897 2,287 2,097 2,897 2,897 2,287 2,287 2,097 2,897 2,897 2,287 2,287 2,076 2,897 2,897 2,287 2,076 2,897 2,849 2,287 2,092 288 0.875 2.569 289 1.544 2.572 291 0.728 2.561 292 0.690 2.032 293 1.096 2.466 294 0.861 2.228 295 1.631 2.696 297 1.598 2.764 298 1.204 2.636 300 1.854 2.636 301 0.796 2.431 302 1.065 2.623 303 0.733 2.381 305 1.375 2.686 306 1.7 2,687 312 1,291 2,553 313 1,100 2,152 314 0.456 1,266 317 0.669 1,051 318 1,329 1,616 319 1,255 2,008 320 1,024 1,941 321 1,493 2,347 322 1,092 1,850 323 1,292 2,077 324 0.281 1,299 330 0.725 1,483 332 0,620 1,381 33 1,258 2,166 340 1,194 1,689 468 181 TABLE 2 (continued) Antigen Serum Test no. v (39) gp41A5 341 1,290 1,763 357 1,138 1,754 358 0.738 1,733 359 0.599 1,033 5 360 0.528 0.840 363 0.711 1,291 365 1,390 1,963 366 0.821 1,682 368 0.665 1.417 10 371 1,183 1,585 372 1,075 1,593 373 0.799 1,711 374 1,046 1,650 375 0.280 0. 377 1,065 1,480 378 1,261 1,644 379 0.610 1,242 380 1,205 1,763 381 1,321 1,645 20 383 0.552 1,146 '384 0.620 1,029 385 0,973 1,618 386 1,687 1,646 387 0.695 1,189 25 388 0.928 1,396 389 0.938 1.544 390 _; 0.679 1.190 391 "" : 709 1,250 392 0.629 1,014 30 393 0.195 0.874 394 0.622 1.244 395 0.799 1.404 396 0.736 1.416 398 0.766 1.647 35 400 1.210 1.621 401 0.482 1.073 402 0.614 1.302 403 0.498 1.145 406 1.570 1.600 40 407 0.742 1.186 408 0.398 0.809 411 1. 0.864 413 0.557 »1.220 45 414 1.170 1.787 416 0.526 1.326 417 0.473 1.201 419 1.020 1.592 420 0.637 0.944 S0 422 0.742 1.479 10 15 20 25 30 35 40 45 S0 Af TABLE 2 (cont -1 -1 388 181 Antiggn Serum Sample No. V (39 ) gp41A5 426 0.921 1.381 427 0.548 1.116 428 0.355 1.184 429 0 .381 1,051 431 0.542 1,198 432 1,192 1,633 433 1,402 1,793 434 0.637 1,196 435 1,017 1,951 436 1,155 1,512 437 0.465 1,408 438 0.517 0.572 439 0.483 0.422 440 1.345 1.380 441 0.618 1.238 442 0.660 1.088 443 0.638 1.288 44 0.662 447 2,737 2,913 448 2,150 2,936 449 1,580 2,901 450 2,287 2,956 451 1,645 2,777 452 1,741 2,636 453 1,998 2,922 454 1,756 2,860 455 2,590 3,010 456 1,022 2,936 457 1,055 2,724 458 1,706 2,868 459 1,631 2,836 2,748 2,749 1,034 1,536 464 2,027 2,936 477 0,586 1,843 478 0.651 1,783 480 0.678 1,554 510 0.807 2,180 512 1,583 2,670 513 1,592 ~ 2,796 514 1,922 2,812 S15 2,102 2,769 516 1,434 1,918 517 0.369 0.791 518 1,143 2,498 520 189 2 5 cont.) / 6 Antíqen 5ef “mPr ° V nr 'v (39) gp41As 521 2,418 2,831 s22 1,107 2,493 s23, _ ._ 0.509 1,043 624 0.026 2,621 szs 0.029 1,993 626 1,456 2,612 s27 2,311 2,049 sza 0.679 2,026 529 0,530 2,631 6 1.6ss 2.746 631 1.177 2. 739 532 0.304 1,617 s33 1,005 2,690 534 1.as2 2,034 s3s 1,412 2,744 536 0.350 1,093 537 0.905 2,764 s3a 1,147 2,590 539 0.219 0.407 540 2,644 0.653 \ (if) / 7 TABLE 3 468 181 ELISA tests for comparison of specificity synthetic peptide antigens 5 10 15 20 25 30 35 40 45 50 55 f Spectrophotometric readings, OD

H Ickë "HIV'-l antigen 405 Antíggn SERUM Negativ PROV NR. WB gp4lAS V (39) antigen ** 4 + 2.809 * 0.436 * 0.084 * 7 + 1.412 0.137 0.066 12 + 2.222 0.336 0.087 19 + 2.922 0.289 0.071 25 + 2.635 0.484 0.071 31 + 2.343 0.737 0.101 40 + 1.495 0.227 0.090 42 + 2.289 0.731 0.140 43 + 1.978 0.522 0.062 44 + 2.443 1.199 0.085 48 + 2.592 0.195 0.073 60 + 1.399 0.219 0.084 78 + 1.826 0.489 0.219 101 + 0.863 0.205 0.072 482 + 1.316 0.120 0.100 511 + 1.036 0.248 0.067 140 + 2.369 0.952 U.081 277 + 2.243 1.106 0.260 285 + 1.977 0.437 0.057 298 + 2.636 1.204 0.046 324 + 1.299 0.281 0.057 375 + 0.957 0.280 0.066 393 + 0.874 0.195 0.067 416 + 1.326 0.526 0.082 42 + 1,184 0.355 0.142 446 + 0.612 1.731 0.215 457 + 2.724 1.055 0.205 477 + 1.843 0.586 0.104 478 + 1.783 0.651 0.069 517 + 0.791 0.369 0.029 520 + 1.923 0.593 0.088 523 + 1.093 0.509 0.080 524 + 2.621 0.825 0.056 525 + 1.993 0. + 1,617 0.384 0.082 536 + 1,093 0.350 0.086 539 + 0.407 0.219 0.051 540 + 2.644 0.643 0.056 39508 ~ 0.154 0. 153 0.087 39509 0.191 0.166 0.043 39510 0.156 0.085 0.044 39511 - 0.241 0.177 0.028 39512 - 0.101 0.144 0.093 39513 0.178 0.123 0.048 39514 0.101 0.118 0.051 39515 - 0.086 0.252 0.063 468 181 'X Areas of use according to the invention gp41A be used as antigen in diagnostic tests regardless of test methodology to detect antibodies to HIV-1. either in solid phase or in competition assay. It can be used as an antigen, possibly linked to any carrier structure, for the immunization of animals to induce the production of antibodies specific for HIV gp41. These antibodies can be used for the detection of HIV-1 or parts of HIV-1 in cell cultures. blood, organ sections, etc .. and partly in competition tests to detect antibodies to HIV-1 or parts of HIV-1. It can also be used to detect antibody-reduced cells and cell-mediated immunity.

A further possible use is as antigen, possibly linked to some carrier structure, to induce the production of antibodies to HIV-1 in animals and humans. m fl

Claims (4)

468 181 I? PATENT REQUIREMENTS A synthetic antigen in the form of a peptide, the sequence of which corresponds to portions of the proteins of the HIV-1 virus, characterized in that the peptide has the following formula: Asp-Gln-Gln-Leu-Leu-Gly-Ile- Trp-Gly-Cys-Ser-Gly-Lys-Leu-Ile-Cys- Thr-Thr-Ala ~ Val-Pro-Trp-Asn-Cys-OH_ A diagnostic test for the detection of antibodies to HIV-1 virus in animals or humans, and which test contains either a peptide, the sequence of which corresponds to portions of the protein of the HIV-1 virus, characterized in that the peptide has the formula given in claim 1. A vaccine for inducing immunity to HIV-1 virus in humans and animals. and which vaccine as active component contains a peptide, the sequence of which corresponds to parts of the proteins of the HIV-1 virus, characterized in that the peptide has the formula stated in claim 1. An antigen intended to induce the production of antibodies to HIV-1 in animals, and which antigen as active ingredient contains a peptide. whose sequence corresponds to parts of the proteins of the HIV-1 virus, characterized in that the peptide has the formula given in claim 1.