EP1425391A1

EP1425391A1 - Therapeutic vaccination method, mutated peptides of hiv reverse transcriptase and their use for vaccination and diagnostic purposes

Info

Publication number: EP1425391A1
Application number: EP01969902A
Authority: EP
Inventors: Brigitte Autran; Assia Samri; Patrice Debre; Vincent Calvez; Christine Katlama; Gaby Haas
Original assignee: Universite Pierre et Marie Curie Paris 6
Current assignee: Universite Pierre et Marie Curie Paris 6
Priority date: 2001-09-14
Filing date: 2001-09-14
Publication date: 2004-06-09
Also published as: JP2005503418A; CA2460377A1; WO2003025166A1; US20050074463A1

Abstract

The invention concerns treatment of infectious and tumoral pathologies comprising a anti-infective and/or anti-tumoral chemotherapeutic treatment phase inducing resistance mutations and a therapeutic vaccination phase directed against said resistance mutations and the agents used in said treatment. More particularly, the invention concerns peptides of 8 to 80 amino acids of the HIV reverse transcriptase sequence and comprising at least a mutation with respect to said wild sequence of said enzyme, mutation induced in response to treatments by nucleoside and non-nucleoside analogues of the HIV reverse transcriptase. The invention also concerns a pharmaceutical composition or vaccine based on said peptides, for inducing an immune response specific of said mutated sequences and for enhancing or prolonging the efficiency of treatments with nucleoside or non-nucleoside analogues of the HIV reverse transcriptase. The invention further concerns epitopes derived from said peptide sequences to evaluate the specific immune response following the vaccine injection.

Description

Therapeutic vaccination methods, mutated HIV reverse transcriptase peptides and their use for vaccination and diagnostic purposes

The subject of the present invention is a treatment of infectious and tumor pathologies comprising a phase of treatment by anti-infectious and / or anti-tumor chemotherapy inducing resistance mutations and a phase of therapeutic vaccination directed against these resistance mutations and to the agents used. as part of this processing.

A more particular subject of the present invention is peptides of 8 to 80 amino acids of the HIV reverse transcriptase sequence and comprising at least one mutation relative to the wild-type sequence of this enzyme, mutation induced in response to treatments with nucleoside or non-nucleoside analogs of HIV reverse transcriptase.

The present invention also relates to a pharmaceutical composition or vaccine based on these peptides, in order to induce a specific immune response of these mutated sequences and to reinforce or prolong the effectiveness of the treatments with nucleoside or non-nucleoside analogues of the HIV reverse transcriptase. .

The present invention also relates to epitopes derived from these peptide sequences in order to evaluate the specific immune response following the injection of a vaccine.

Identifying an effective HIV vaccine is critical. Effective immunization would be effective, affordable at a monetary level, and would be a long-term approach to this infection. However, the biggest obstacle to developing effective vaccines is the variability of HIV. Current data suggest that cytotoxic T lymphocytes (CTLs) are responsible for controlling the initial peak of viremia, low-noise viral replication during the asymptomatic phase of the disease, and the elimination of most viral variants .

The recognition of an infectious agent as a foreign supposes that the immune system recognizes certain specific foreign structures, the antigens, constituting the non-self, distinguishing them from the structures which belong to it, constituting the self. The immune response actors involve:

* the lymphocytes which are the effector cells of the immune reaction. They include B and T lymphocytes subdivided into two subpopulations: CD4 + helper T cells (Th), coordinators of specific immune responses, and CD8 + cytotoxic T lymphocytes (CTLs). These CTLs are capable of recognizing and killing cells infected with viruses, in particular, before the virus buds on the surface of the cell and is released into the extracellular medium. * the antigen presenting cells which capture the antigens, condition them and present them in immunogenic form to the T lymphocytes.

* MHC molecules (major histocompatibility complex) which are expressed on the surface of cells and participate in the presentation of the antigen to lymphocytes. In humans, MHC is called HLA (human leukocyte antigen). There are two classes of MHC molecules: class I molecules which are present on the surface of all nucleated cells in the body and class II molecules which are only expressed on the surface of cells presenting antigens. Antigenic peptides of 8 to 10 amino acids bind in a niche of MHC class I molecules, peptides of 14-15 amino acids bind to MHC class I Their binding is determined by the interaction forces established between the amino acids (essentially in the two terminal parts) of the antigenic peptide and those of the walls of the niche of HLA molecules. More than 200 HLA class I alleles have now been defined within the three families of HLA class IA genes, B and C. Each HLA class I allele is significantly different in terms of the peptide niche, so the peptides that can s 'fix it are also different. There is therefore a strict specificity in the interaction between peptide and HLA class I molecule. This specificity explains why even a minor change in the sequence of the peptide presented as target may be sufficient to make this peptide incapable of binding to the HLA molecule class I. * receptors for the antigen which are fixed in the membrane of T lymphocytes

Thanks to this receptor, each T lymphocyte specifically recognizes an antigen presented on the MHC. The receptor for T lymphocytes is called TCR (T Cell Receptor). This receptor must recognize at the same time on the cell surface the antigenic peptide and the MHC molecule. It is said that recognition is restricted by the CMH.

* CD markers (differentiation cluster) which are molecules whose presence on the cell membrane identifies a cell. Thus, the Th carry the CD4 marker and are also called CD4 + and the CTLs which carry the marker

CD8 are called CD8 +. CD8 + lymphocytes recognize their antigen on MHC class I; CD4 + lymphocytes on MHC class IL

* The presentation of the antigenic sequences by the antigen presenting cells is one of the determining factors of the effectiveness of a vaccine. It can be amplified by certain natural or synthetic compounds, adjuvants, such as lipids (fatty acids, phospholipids, Freund's adjuvant), anionic copolymers, CpG units ... Other molecules such as cytokines (interleukins , interferons, TNF (tumor necrosis factors), TGF (transforming growth factors ...) and chemokines are involved in the regulation of maturation, activation, proliferation and differentiation of cells of the immune system, and play a major role in the molecule of vaccine efficacy.

HIV infection is an infection of the immune system in the sense that this virus electively infects CD4 + lymphocytes and antigen-presenting cells and gradually destroys the immune system, usually over 10 years. Some people who have been infected for more than 20 years and who have never been treated with antiretroviral therapies, however, have a very low viral load (or below the detection limit) and present very important virus-specific CTLs responses, directed against epitopes. restricted to HLA class I molecules. The intensity and diversity of these CTL responses to HIV could be correlated with the control of viral replication, suggesting the crucial role played by these CTLs in these immune defenses.

CTL target HIV peptide sequences have been defined for the majority of HIV proteins. These targets differ depending on the individual, depending on the HLA class I molecules expressed on the surface of the cells of a given individual.

The escape of the virus from the control exerted by the immune system is the main cause of the ineffectiveness of these responses. Viral variation in CTL epitopes may play a key role in the immune system's failure to contain the virus. A single mutation in a defined epitope recognized by a CTL may indeed be sufficient to suppress CTL recognition, or to block the binding of the peptide to the class I molecule, or by altering residues essential for interactions with the TCR. The sequence of the HIV Reverse Transcriptase enzyme being highly conserved between the different virus subtypes, it is a prime target for CTL-type immune responses: 80% of the patients tested exhibit a CTL response directed against reverse transcriptase corresponding to HIV-1.

A large number of HIV vaccines have been developed and tested. These vaccines include recombinant VLH proteins, synthetic peptides, recombinant viral or bacterial vectors, naked DNA vaccines. More than 60 phase I clinical trials on thirty candidate specific type B vaccines for HIV, which is the most common subtype in the United States and Europe, have been carried out or are underway. Most of these trials have focused on the HIV envelope protein. However, the recombinant proteins tested rarely induce CD8 + CTLs which recognize and kill cells infected with HIV. In addition, type I HIV isolates exhibit a degree of intense genetic diversity in the viral envelope that can affect aspects of the life cycle such as infectivity, transmissibility or immunogenicity. AIDS treatments use molecules capable of blocking one or more stages of the virus cycle without altering the proper functioning of the host cells. The other structural proteins (the viral capsid and reverse transcriptase, in particular) as well as the regulatory proteins, which are more conserved than the viral envelope, are today the preferred targets of preventive vaccination strategies.

* Reverse transcriptase (RT), a viral polymerase that catalyzes the synthesis of viral DNA from the RNA genome and is a prime target for antiretroviral treatments. Non-hydrolyzable nucleoside analogs (NRTI) such as AZT (zidovudine), DDI (didanosine), DDC (zalcitabine), 3TC (lamivudine), ABC (abacavir), D4T (stavudine) ... constitute the family of inhibitors of the most used RT to date. In triphosphorylee form, they compete with natural nucleosides during DNA synthesis and cause the termination of its elongation. Another family of reverse transcription stage inhibitors, non-nucleoside inhibitors of the enzyme (NNRTI), does not require phosphorylation to be active and probably acts by allosteric inhibition. The best known are Nevirapine, Delavirdine or Efavirenz.

* The HIV protease is the other main target of anti-retroviral treatments that use enzyme inhibitors such as Ritonavir,

Saquinavir, Indinavir ...

Current antiretroviral therapies generally combine several classes and include one or more reverse transcriptase inhibitors. However, these treatments cannot eradicate the virus and the residual replication which persists during these treatments, even the most active, allows the selection over time of mutations in the reverse transcriptase gene. Some of these mutations give the mutated viras the ability to resist the inhibitory action of these drugs. Thus many mutations affect in particular the reverse transcriptase sequence (including changes in the active site of the enzyme), thus favoring the emergence of variant viruses resistant to the families of NRTIs or

NNRTI within one to twelve months of processing. The accumulation of these mutations over time in patients treated with these drugs leads to the selection of multidrug-resistant viruses and leads to therapeutic failures.

It has been shown in patent application WO99 / 51750 that treatment with Lamivudine induces point mutations in the catalytic site of HIV reverse transcriptase. This point mutation is located in the site of recognition of the peptide by the TCR. As a result, the CTLs of the treated patient who recognized a wild type epitope were no longer able to recognize a mutated epitope. The latest estimates show that, of the 85% of patients with

HIV treated with at least one reverse transcriptase inhibitor in Western countries, 50% of them maintain a detectable viremia and are considered to be in therapeutic failure. This is associated with the existence of at least one resistance mutation to these inhibitors and results in an increase in the viral load and a decrease in CD4 + T lymphocytes. In addition, 10% of patients recently infected, but not yet treated, are infected with new viral strains carrying mutations in resistance to antiretroviral treatment. Numerous studies have shown that subsequent antiviral therapies with molecules of the same class then have less effectiveness on these resistant viruses. An alternative therapeutic strategy is now proposed in order to limit the toxicity and the cost of these long-term treatments. The aim is to stimulate the immune responses to HIV in treated patients by combining these treatments with anti-HIV immunization by the available vaccine candidates in order to discontinue the treatments in the long term. Phase I and II clinical trials, using vaccine compounds directed against the consensus consensus sequences of non-mutated viruses, are in progress.

The Applicant has discovered a treatment for infectious and tumor pathologies comprising a phase of treatment with anti-infectious and / or anti-tumor chemotherapy inducing resistance mutations and a phase of therapeutic vaccination directed against these resistance mutations.

The Applicant has discovered more particularly that certain sequences of reverse transcriptase sites of point and standardized mutations occurring during various treatments with NRTI and / or NNRTI, are in fact capable of causing the induction of specific CTLs capable of recognizing these mutations . It has been demonstrated ex vivo, by ELISPOT type methods, the existence of CD8 + cells (of the order of 50 to 500 specific cells per million blood mononuclear cells) directed against several mutations of the reverse transcriptase induced by NRTIs, such as the mutation 41, 74, 184 .... within the mononuclear blood cells of the peripheral blood of infected, treated and carriers of these mutations. This immune response can therefore take place during these treatments, but seems incapable of ensuring the elimination of specific variants on its own. Amplification of the immunity directed against these mutations will make it possible to control the emergence of these variants of HIV and to maintain a lasting viral suppression under these treatments. The administration of vaccines containing these mutated reverse transcriptase viral sequences, before or during the administration of NRTI or NNRTI treatments should make it possible to prevent, delay or reduce the appearance of mutated viruses capable of resisting these treatments. This new type of therapeutic immunization thus aims to strengthen or prolong the effectiveness of antiretroviral treatments with NRTIs and / or NNRTIs. This can be secondarily applied to other mutations induced by other classes of antiretrovirals and can also be used in preventive vaccination. The invention therefore consists in a treatment of infectious and / or tumor pathologies.

The invention also consists of the treatment agents used in the context of this treatment. The subject of the invention is also the preparation of immunogens containing the mutations of resistance to NRTI and / or NNRTI.

More particularly, the invention consists in the preparation of immunogens containing the resistance mutations to one or more NNRTIs used alone or in conjunction with one or more NRTIs. The subject of the invention is also the peptides which are elements of immunogens.

Another subject of the invention is a pharmaceutical composition characterized in that it comprises at least one peptide as defined, dispersed in pharmaceutically acceptable excipients, this composition being used as a vaccine. Another object of the invention is the determination of epitopes recognized by the

CTLs specific for vaccine peptide sequences introduced for vaccination, usable for diagnostic purposes.

The method for treating infectious and / or tumor pathologies comprises a phase of treatment by anti-infectious and / or antitumour chemotherapy inducing resistance mutations and a phase of therapeutic vaccination directed against these resistance mutations. Preferably, the vaccination phase follows the chemotherapy treatment phase.

The present invention also relates to an agent for treating infectious and / or tumor pathologies which comprises at least two components:

the first component comprising a medicament intended for the treatment of infectious and or anti-tumor pathologies inducing resistance mutations,

- the second component comprising a vaccine directed against these resistance mutations, the two components being intended for simultaneous, separate or sequential use over time.

The present invention also relates to peptides of 8 to 80 amino acids of the HIV reverse transcriptase sequence comprising at least one mutation by substitution and / or by single or multiple insertion with respect to established reverse transcriptase sequences, in the absence of any treatment, mutation observed following treatment with NRTI and / or NNRTI, the peptides being characterized by the fact that they are capable of inducing a response mediated by T lymphocytes specific for this mutated sequence of HIV reverse transcriptase, with the exception of peptides of sequence Z _r YVDDZ ₂ , Z _r YIDDZ ₂ and Z _r

YLDD-Z2 with Z _x and Z ₂ being at least any amino acid, these peptides having been observed following treatment with lamivudine alone (NRTI).

In particular, the subject of the invention is preferably peptides of 8 to 80 amino acids of the HIV reverse transcriptase sequence comprising at least one mutation by substitution and / or by single or multiple insertion with respect to the established sequences of the reverse transcriptase, in the absence of any treatment, mutation observed following treatment with NNRTI alone or following treatment combining NNRTI and NRTI, the peptides being characterized by the fact that they are capable of induce a response mediated by T lymphocytes specific for this mutated sequence of VLH reverse transcriptase.

Preferably, the peptides according to the invention contain from 15 to 50 amino acids.

The sequences of the peptides according to the invention can be preceded or followed by neutral amino acids which can dissolve or stabilize the peptides. The peptides according to the invention are characterized by the sequences or parts of sequence comprising at least one of the following Mp mutation with Xn being an amino acid corresponding to that described in the Los Alamos database at the same position of the peptide sequence of HIV-1 reverse transcriptase, and n being the position of an amino acid described in the Los Alamos database in the sequence of HIV-1 reverse transcriptase, and with Mp being a mutated amino acid and p being the number of the mutation observed.

* Sequence A: aa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

X X X X X Ml X X X X X X X X X in which Ml (aa 6) can be mutated into K, G or S

* Sequence B: aa 30 3 1 32 33 34 35 36 37 38 39 40 41 42 43 44

XXXXXXXXX M2 X M3 XXX 45 46 47 48 49 50 XXXXXX in which M2 (aa 39) can be X39 or be mutated into A, K, N, P, M or S;

M3 (aa 41) can be X41 or be mutated to L; at least one of M2 and M3 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention, M2 (aa 39) can be mutated to A, K, N, P, M or S;

M3 (aa 41) can be X41 or be mutated to L;

* Sequence C aa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

X X X X X Ml X X X X X X X X X X

16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

X X X X X X X X X X X X X X X

31 32 33 34 35 36 37 38 39 40 41 42 43 44 45

X X X X X X X X M2 X M3 X X X X

46 47 48 49 50 51

X X X X X X in which Ml (aa 6) can be X6 or be mutated into K, G or S;

M2 (aa 39) can be X39 or be mutated to A, K, N, P, M or S;

M3 (aa 41) can be X41 or be mutated to L; at least one of Ml, M2 and M3 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention, Ml (aa 6) can be X6 or be mutated into K, G or S;

M2 (aa 39) can be X39 or be mutated to A, K, N, P, M or S;

M3 (aa 41) can be X41 or be mutated to L; with Ml or M2 representing an amino acid carrying a mutation.

* Sequence D

53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 aa

XXXXXXXXX M4 XX M5 X M6 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82

M7 M8 M9 X X X M10 Mi l X M12 X X X X X

83 84 85 86 X X X X in which M4 (aa 62) can be X62 or be mutated into V; M5 (aa 65) can be X65 or be mutated to R; M6 (aa 67) can be X67 or be mutated to N or G; M7 (aa 68) can be X68 or be mutated to G or N; M8 (aa 69) can be X69 or be mutated to D, N, S, A, or combined with multiple inserts like SG, SX, SSA, SSG, SSS, SEA, STS, ASG, SXX, or XXX;

M9 (aa 70) can be X70 or be mutated to R, N, E or S; M10 (aa 74) can be X74 or be mutated into V or I; Ml 1 (aa 75) can be X75 or be mutated to M, I, L or T;

M12 (aa 77) can be X77 or be mutated to L; at least one of M4 to Ml 2 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention, M4 (aa 62) can be X62 or be mutated to V; M5 (aa 65) can be X65 or be mutated to R;

M6 (aa 67) can be X67 or be mutated to N or G;

M7 (aa 68) can be X68 or be mutated to G or N;

M8 (aa 69) can be X69 or be mutated to D, N, S, A, or combined with multiple inserts like SG, SX, SSA, SSG, SSS, SEA, STS, ASG, SXX, or XXX;

M9 (aa 70) can be X70 or be mutated to R, N, E or S;

M10 (aa 74) can be X74 or be mutated into V or I;

Mil (aa 75) can be X75 or mutated to M, I, L or T;

M12 (aa 77) can be X77 or be mutated to L; at least one of M4, M7 or M12 representing an amino acid carrying a mutation.

Sequence E

51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 aa

XXXXXXXXXXX M4 XX M5 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 X M6 M7 M8 M9 XXX M10 Mil X M12 XXX

81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 X X X X X X X M13 M14 M15 X X X X X

96 97 98 99 100 101 102 103 104 105 106 107 108 109 1 10 X X M16 X M17 M18 X M19 X X M20 X M21 X X

1 1 1 1 12 1 13 1 14 115 116 1 17 1 18 119 120 121 122 123 124 125 X X X X M22 M23 X X M24 X X X X

126 127 128 X X X in which M4 (aa 62) can be X62 or be mutated into V;

M5 (aa 65) can be X65 or be mutated to R;

M6 (aa 67) can be X67 or be mutated to N or G;

M7 (aa 68) can be X68 or be mutated to G or N;

M9 (aa 70) can be X70 or be mutated to R, N, E or S;

M10 (aa 74) can be X74 or be mutated into V or I;

Mil (aa 75) can be X75 or mutated to M, I, L or T;

M12 (aa 77) can be X77 or be mutated to L;

M13 (aa 88) can be X88 or be mutated to C;

M14 (aa 89) can be X89 or be mutated to G;

M15 (aa 90) can be X90 or be mutated to I;

M16 (aa 98) can be X98 or be mutated into G or S;

M17 (aa 100) can be X100 or be mutated to I;

M18 (aa 101) can be X101 or be mutated to E, Q, R, I or P;

M19 (aa 103) can be X103 or be mutated to N, Q, R, T or S;

M20 (aa 106) can be X106 or be mutated to A, I or L;

M21 (aa 108) can be X108 or be mutated to I;

M22 (aa 115) can be XI 15 or be mutated to F;

M23 (aa 116) can be XI 16 or be mutated to Y;

M24 (aa 119) can be XI 19 or be mutated to S; at least one of M4 to M24 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention, M4 (aa 62) can be X62 or be mutated to V;

M5 (aa 65) can be X65 or be mutated to R;

M6 (aa 67) can be X67 or be mutated to N or G;

M7 (aa 68) can be X68 or be mutated to G or N;

M9 (aa 70) can be X70 or be mutated to R, N, E or S;

M10 (aa 74) can be X74 or be mutated into V or I;

Ml 1 (aa 75) can be X75 or be mutated to M, I, L or T;

M12 (aa 77) can be X77 or be mutated to L;

M13 (aa 88) can be X88 or be mutated to C;

M14 (aa 89) can be X89 or be mutated to G;

Ml 5 (aa 90) can be X90 or be mutated to I;

Ml 6 (aa 98) can be X98 or be mutated to G or S;

M17 (aa 100) can be X100 or be mutated to I;

M18 (aa 101) can be X101 or be mutated to E, Q, R, I or P;

M19 (aa 103) can be X103 or be mutated to N, Q, R, T or S;

M20 (aa 106) can be X106 or be mutated to A, I or L;

M21 (aa 108) can be X108 or be mutated to I;

M22 (aa 115) can be XI 15 or be mutated to F;

M23 (aa 116) can be XI 16 or be mutated to Y;

M24 (aa 119) can be XI 19 or be mutated to S; at least one of M4, M7, M12 to M21, M23 or M24 representing an amino acid carrying a mutation.

Sequence F aa 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143

X X X X X X X X X M25 M26 X M27 X X

144 145 ι 6 147 148 149 150 151 152 153 154 155 156 157 158

XXXXXXX M28 XXXXX M29 X 159 160 161 162 163 164 165 166 XXXXXXXX in which M25 (aa 138) can be XI 38 or be mutated into A, G, K or Q; M26 (aa 139) can be X139 or be mutated to I, M or K; M27 (aa 141) can be X141 or be mutated to E; M28 (aa 151) can be X151 or be mutated to M; M29 (aa 157) can be X157 or be mutated to S; at least one of M25 to M29 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention,

M25 (aa 138) can be X138 or be mutated to A, G, K or Q; M26 (aa 139) can be X139 or be mutated to I, M or K;

M27 (aa 141) can be X141 or be mutated to E;

M28 (aa 151) can be X151 or be mutated to M;

M29 (aa 157) can be X157 or be mutated to S; at least one of M25 to M28 representing an amino acid carrying a mutation.

Sequence G

1 63 164 165 166 167 168 169 170 171 172 173 174 175 176 177 aa

X X X X X X X X X M30 X X X X X

178 179 180 181 182 183 184 185 1 86 187 188 189 190 191 192 M31 M32 X M33 X X M34 X X X M35 M36 M37 X X

193 194 195 196 197 198 199 X X X X X X X in which M30 (aa 172) can be X172 or be mutated to K;

M31 (aa 178) can be X178 or be mutated to M, L, or F M32 (aa 179) can be X179 or be mutated to I, L, D or E;

M33 (aa 181) can be X181 or be mutated to C or I;

M34 (aa 184) can be X184 or be mutated to V, I or T;

M35 (aa 188) can be X188 or be mutated into L, C or H;

M36 (aa 189) can be XI 89 or be mutated to I; M37 (aa 190) can be X190 or be mutated to A, S, E, Q or T; at least one of M30 to M37 representing an amino acid carrying a mutation. In a preferred embodiment of the invention, M30 (aa 172) can be X172 or be mutated to K; M31 (aa 178) can be X178 or be mutated to M, L, or F M32 (aa 179) can be X179 or be mutated to I, L, D or E; M33 (aa 181) may be XI 81 or be mutated to C or I;

M34 (aa 184) can be XI 84 or be mutated to V, I or T; M35 (aa 188) can be XI 88 or be mutated into L, C or H; M36 (aa 189) can be XI 89 or be mutated to I; M37 (aa 190) can be X190 or be mutated to A, S, E, Q or T; at least one of M30, M33, M35 to M37 representing an amino acid carrying a mutation.

* H sequence

169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 aa X X X M30 X X X X X M31 M32 X M33 X X

184 185 M34 X in which M30 (aa 172) may be X172 or be mutated to K;

M31 (aa 178) can be X178 or be mutated to M, L, or F;

M32 (aa 179) can be X179 or be mutated into I, L, D or E;

M33 (aa 181) can be X181 or be mutated to C or I;

M34 (aa 184) can be X184 or be mutated to V, I or T; at least one of M30 to M34 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention,

M30 (aa 172) can be X172 or be mutated to K;

M31 (aa 178) can be X178 or be mutated to M, L, or F; M32 (aa 179) can be X179 or be mutated into I, L, D or E;

M33 (aa 181) may be XI 81 or be mutated to C or I;

M34 (aa 184) can be XI 84 or be mutated to V, I or T; at least one of M30; M32 or M33 representing an amino acid carrying a mutation. Sequence I

184 1 85 186 187 188 189 190 191 192 193 194 195 196 197 198 199 M34 XXX M35 M36 M37 XXXXXXXXX in which M34 (aal84) can be XI 84 in which M35 (aa 188) can be X188 or be mutated in L, C or H; M36 (aa 189) can be XI 89 or be mutated to I;

M37 (aa 190) can be X190 or be mutated to A, S, E, Q or T; at least one of M34 to M37 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention, M34 (aal 84) can be XI 84 in which M35 (aa 188) can be X188 or be mutated into L, C or H;

M36 (aa 189) can be XI 89 or be mutated to I;

M37 (aa 190) can be X190 or be mutated to A, S, E, Q or T; at least one of M35 to M37 representing an amino acid carrying a mutation.

Sequence J

aa 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 X X X X X X X X X M38 X M39 M40 X X

214 215 216 217 21 8 219 220 221 222 223 224 225 226 227 228 M41 M42 X X X M43 X X X M44 X M45 X M46 M47

229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 X X X X M48 M49 X M50 X M51 X M52 X X X

244 245 246 247 248 249 X X X X X X in which M38 (aa 208) may be X208 or be mutated to Y;

M39 (aa 210) can be X210 or be mutated to W; M40 (aa 211) can be X211 or mutated to K, A, E, N, D, G or Q;

M41 (aa 214) can be X214 or be mutated into L or F;

M42 (aa 215) can be X215 or be mutated to Y, F or C;

M43 (aa 219) can be X219 or be mutated to Q, E N, R, T or W;

M44 (aa 223) can be X223 or be mutated to Q; M45 (aa 225) can be X225 or be mutated to H; M46 (aa 227) can be X227 or be mutated to L; M47 (aa 228) can be X228 or be mutated to R, H or F; M48 (aa 233) can be X233 or be mutated to V or N; M49 (aa 234) can be X234 or be mutated into I, H or P; M50 (aa 236) can be X236 or be mutated to M, S or L;

M51 (aa 238) can be X238 or be mutated to T or S; M52 (aa 240) can be X240 or be mutated to A, D or I; at least one of M38 to M52 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention,

M38 (aa 208) can be X208 or be mutated to Y;

M39 (aa 210) can be X210 or be mutated to W;

M40 (aa 211) can be X211 or mutated to K, A, E, N, D, G or Q;

M41 (aa 214) can be X214 or be mutated into L or F; M42 (aa 215) can be X215 or be mutated to Y, F or C;

M43 (aa 219) can be X219 or be mutated to Q, E N, R, T or W;

M44 (aa 223) can be X223 or be mutated to Q;

M45 (aa 225) can be X225 or be mutated to H;

M46 (aa 227) can be X227 or be mutated to L; M47 (aa 228) can be X228 or be mutated to R, H or F;

M48 (aa 233) can be X233 or be mutated to V or N;

M49 (aa 234) can be X234 or be mutated into I, H or P;

M50 (aa 236) can be X236 or be mutated to M, S or L;

M51 (aa 238) can be X238 or be mutated to T or S; M52 (aa 240) can be X240 or be mutated to A, D or I; at least one of M38, M40, M44 to M52 representing an amino acid carrying a mutation.

Sequence K

324 325 326 327 328 329 330 331 332 333 334 335 336 337 335 X X X X X X X X X M53 X X X X X

339 340 341 342 XXXX in which M53 (aa 333) can be mutated into E or D; Preferably, the peptides according to the invention are characterized by the following sequences or parts of sequence comprising at least one of the following Mp mutations:

* Sequence A bis:

AA 1 2 3 4 5 6 7 8 9 10 1 1 12 13 14 15 P I S P I M1 T V P V K L K P G in which Ml (aa 6 - wt = E) can be mainly transferred to K, or to G or S;

equιenoeB bis:

AA 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44

K 1 K A L V E I C M2 E M3 E K E

45 46 47 48 49 50

G K 1 S K I in which M2 (aa 39- wt = T) can be X39 or be mutated mainly in A, or in K, or N, or P, or M or S;

M3 (aa 41- wt = M) can be X41 or be transferred mainly to L; at least one of M2 and M3 representing an amino acid carrying a mutation.

M3 (aa 41) can be X41 or be mutated to L;

* Sequence C bis:

AA 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

P 1 S P 1 Ml T V P v K L K P G

16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

M D G P K V K Q W P L T E E K

31 32 33 34 35 36 37 38 39 40 41 42 43 44 45

1 K A L V E 1 C M2 E M3 E K E G

46 47 48 49 50 51

K 1 SK 1 G in which Ml (aa 6- wt = E) can be mainly transferred to K, or to G or S M2 (aa 39- wt = T) can be X39 or be transferred mainly to A, or to K, or N, or P, or M or S;

M3 (aa 41- wt = M) can be X41 or be mainly transferred to L; at least one of Ml, M2 and M3 representing an amino acid carrying a mutation.

M2 (aa 39) can be X39 or be mutated to A, K, N, P, M or S; M3 (aa 41) can be X41 or be mutated to L; with Ml or M2 representing an amino acid carrying a mutation.

* Sequence DA:

AA 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67

E N P Y N T P V F M4 1 K M5 K M6

68 69 70 71 72 73 74 75 76 77 78 79 80 81 82

M7 M8 M9 W R K M10 Mil D M12 R E L N K

83 84 85 86

R T Q D

in which M4 (aa 62-wt = A) can be X62 or be mutated mainly in V;

M5 (aa 65- wt = K) can be X65 or be mutated mainly in R;

M6 (aa 67- wt = D) can be X67 or be mutated mainly in N or G;

M7 (aa 68- wt = S) can be X68 or be mutated mainly in G or N;

M8 (aa 69- wt = T) can be X69 or be mutated mainly in D, or in N, or S, or A, or combined with multiple insertions like SG, SX, SSA, SSG, SSS,

SEA, STS, ASG, SXX, or XXX;

M9 (aa 70- wt = K) can be X70 or be mutated mainly in R, or in N, or E or S;

M10 (aa 74- wt = L) can be X74 or be mutated mainly in V or in I; Ml 1 (aa 75- wt = V) can be X75 or be mutated mainly in M, or in I, or L or T;

M12 (aa 77- wt = F) can be X77 or be transferred mainly to L; at least one of M4 to M12 representing an amino acid carrying a mutation In a preferred embodiment of the invention, M4 (aa 62) can be X62 or be mutated to V; M5 (aa 65) can be X65 or be mutated to R; M6 (aa 67) can be X67 or be mutated to N or G; M7 (aa 68) can be X68 or be mutated to G or N; M8 (aa 69) can be X69 or be mutated to D, N, S, A, or combined with multiple inserts like SG, SX, SSA, SSG, SSS, SEA, STS, ASG, SXX, or XXX;

Sequence E bis:

51 52 53 54 55 56 57 58 59 60 61 62 63 64 65

G P E N P Y N T P V F M4 I K M5

66 67 68 69 70 71 72 73 74 75 76 77 78 79 80

K M6 M7 M8 M9 W R K M10 Mil D M12 R E L

81 82 83 84 85 86 87 88 89 90 91 92 93 94 95

N K R T Q D F M13 M14 M15 Q L G 1 P

96 97 98 99 100 101 102 103 104 105 106 107 108 109 110

H P M16 G M17 M18 K M19 K S M20 T M21 L D

111 112 113 114 115 116 117 118 119 120 121 122 123 124 125

V G D A M22 M23 S V M24 L D E D F R

126 127 128

K Y T in which M4 (aa 62- wt = A) can be X62 or be mutated mainly in V; M5 (aa 65- wt = K) can be X65 or be mutated mainly in R; M6 (aa 67- wt = D) can be X67 or be mainly mutated into N or G; M7 (aa 68- wt = S) can be X68 or be mutated mainly in G or in N;

M8 (aa 69- wt = T) can be X69 or be mutated mainly in D, or in N, or S, or A, or combined with multiple insertions like SG, SX, SSA, SSG, SSS, SEA, STS, ASG, SXX, or XXX; M9 (aa 70- wt = K) can be X70 or be mutated mainly in R, or in N, or E or S;

M10 (aa 74- wt = L) can be X74 or be mutated mainly in V or in I;

Ml 1 (aa 75- wt = V) can be X75 or be mutated mainly in M, or in I, or L or T;

M12 (aa 77- wt = F) can be X77 or be transferred mainly to L;

M13 (aa 88- wt = W) can be X88 or be mutated mainly in C;

M14 (aa 89- wt = E) can be X89 or be mainly transferred to G;

M15 (aa 90- wt = V) can be X90 or be mainly transferred to I; M16 (aa 98- wt = A) can be X98 or be mainly transferred to S or G;

Ml 7 (aa 100-wt = L) can be XI 00 or be mainly transferred to I;

M18 (aa 101- wt = K) can be X101 or be mainly transferred to E, or to Q, or

R, or I or P;

M19 (aa 103- wt = K) can be X103 or be mutated into N, mainly Q, or into R, or T or S;

M20 (aa 106- wt = V) can be X106 or be transferred mainly to A, or to I or

L;

M21 (aa 108- wt = V) can be X108 or be mutated mainly in I;

M22 (aa 115- t = Y) can be XI 15 or be mainly transferred to F; M23 (aa 116-wt = F) can be XI 16 or be mainly transferred to Y;

M24 (aa 119- t = P) can be XI 19 or be transferred mainly to S; at least one of M4 to M24 representing an amino acid carrying a mutation.

M6 (aa 67) can be X67 or be mutated to N or G;

M7 (aa 68) can be X68 or be mutated to G or N;

M9 (aa 70) can be X70 or be mutated to R, N, E or S;

M10 (aa 74) can be X74 or be mutated into V or I;

Ml 1 (aa 75) can be X75 or be mutated to M, I, L or T;

M12 (aa 77) can be X77 or be mutated to L; M13 (aa 88) can be X88 or be mutated to C; M14 (aa 89) can be X89 or be mutated to G; Ml 5 (aa 90) can be X90 or be mutated to I; M16 (aa 98) can be X98 or be mutated into G or S; M17 (aa 100) can be X100 or be mutated to I;

M18 (aa 101) can be X101 or be mutated to E, Q, R, I or P; M19 (aa 103) can be X103 or be mutated to N, Q, R, T or S; M20 (aa 106) can be X106 or be mutated to A, I or L; M21 (aa 108) can be X108 or be mutated to I; M22 (aa 115) can be XI 15 or be mutated to F;

M23 (aa 116) can be XI 16 or be mutated to Y; M24 (aa 119) can be XI 19 or be mutated to S; at least one of M4, M7, M12 to M21, M23 or M24 representing an amino acid carrying a mutation.

* Sequence F bis: ιA 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 A F T 1 P S I N N M25 M26 P M27 I R

144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 Y Q Y N V L P M28 G W K G S M29 A

159 160 161 162 163 164 165 166 1 F Q S S M T K in which M25 (aa 138- wt = E) can be X138 or be transferred mainly in

A, or G, or K or Q; M26 (aa 139- wt = T) can be X139 or be mainly transferred to I, or to M or

K;

M27 (aa 141-wt = G) can be X141 or be mainly transferred to E;

M28 (aa 151- wt = Q) can be X151 or be mainly transferred to M;

M29 (aa 157-wt = P) can be X157 or be mainly transferred to S; at least one of M25 to M29 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention, M25 (aa 138) can be XI 38 or be mutated into A, G, K or Q; M26 (aa 139) can be X139 or be mutated to I, M or K; M27 (aa 141) can be X141 or be mutated to E; M28 (aa 151) can be X151 or be mutated to M; M29 (aa 157) can be X157 or be mutated to S; at least one of M25 to M28 representing an amino acid carrying a mutation.

* Sequence G bis:

AA 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 S M T K I L E P F M30 K Q N P D

178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 M31 M32 I M33 Q Y M34 D D L M35 M36 M37 S D

193 194 195 196 197 198 199 L E I G Q H R in which M30 (aa 172- wt = R) can be X172 or be transferred mainly in

K;

M31 (aa 178- wt≈I) can be X178 or be transferred mainly to M, L or F; M32 (aa 179- wt = V) can be X179 or be mainly transferred to I, or to L, or

D or E;

M33 (aa 181- wt≈Y) can be XI 81 or be mainly transferred to C or to I;

M34 (aa 184- wt≈M) can be XI 84 or be mainly transferred to V, or to I or

T; M35 (aa 188- wt≈Y) can be X188 or be mainly transferred to L, or to C or

H;

M36 (aa 189- wt = V) can be XI 89 or be transferred mainly to I;

M37 (aa 190- wt≈G) can be X190 or be transferred mainly to A, or to S, or

E, or Q or T; at least one of M30 to M37 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention,

M30 (aa 172) can be X172 or be mutated to K;

M31 (aa 178) can be X178 or be mutated to M, L, or F

M32 (aa 179) can be X179 or be mutated into I, L, D or E;

M33 (aa 181) can be X181 or be mutated to C or I;

M34 (aa 184) can be XI 84 or be mutated to V, I or T;

M35 (aa 188) can be X188 or be mutated into L, C or H;

M36 (aa 189) can be XI 89 or be mutated to I;

M37 (aa 190) can be X190 or be mutated to A, S, E, Q or T; at least one of M30, M33, M35 to M37 representing an amino acid carrying a mutation.

* Sequence H bis:

AA 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 E P F M30 K Q N P D M31 M32 I M33 Q Y

184 185 M34 D in which M30 (aa 172- wt = R) can be X172 or be transferred mainly in

K;

M31 (aa 178-wt = I) can be X178 or be mutated mainly in M, or in L;

M32 (aa 179- wt = V) can be X179 or be mutated mainly in I, or in L, or D or E;

M33 (aa 181-wt≈Y) can be X181 or be mutated mainly in C or in I;

M34 (aa 184- wt = M) can be XI 84 or be mainly transferred to V, or to I or

T; at least one of M30 to M34 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention,

M30 (aa 172) can be X172 or be mutated to K;

M31 (aa 178) can be X178 or be mutated to M, L, or F;

M32 (aa 179) can be X179 or be mutated into I, L, D or E; M33 (aa 181) may be XI 81 or be mutated to C or I;

M34 (aa 184) can be XI 84 or be mutated to V, I or T; at least one of M30; M32 or M33 representing an amino acid carrying a mutation.

* Sequence I bis:

AA 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 M34 D D L M35 M36 M37 S D L E I G Q H

199 R in which M34 (aa 184-wt = M) can be X184 or be mutated mainly in V, or in I or T; in which M35 (aa 188-wt≈Y) can be X188 or be mutated mainly in L, or in C or H;

M36 (aa 189- wt≈V) can be XI 89 or be transferred mainly to I M37 (aa 190- wt≈G) can be X190 or be transferred mainly to A, or S, or E, or Q or T; at least one of M34 to M37 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention, M34 (aal84) can be XI 84 in which M35 (aa 188) can be X188 or be mutated into L, C or H;

M36 (aa 189) can be XI 89 or be mutated to I; M37 (aa 190) can be X190 or be mutated to A, S, E, Q or T; at least one of M35 to M37 representing an amino acid carrying a mutation.

* Sequence J bis:

AA 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 R T K I E E L R Q M38 L M39 M40 W G

214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 M41 M42 T P D M43 K H Q M44 E M45 P M46 M47

229 230 23 1 232 233 234 235 236 237 238 239 240 241 242 243 W M G Y M48 M49 H M50 D M51 W M52 V Q P

244 245 246 247 248 249 I V L P E K in which M38 (aa 208- wt≈H) can be X208 or be transferred mainly in

Y;

M39 (aa 210- wt = L) can be X210 or be mainly transferred to W; M40 (aa 211- wt≈R) can be X211 or be mutated mainly in K, or in A, or E, or N, or D, or G or Q;

M41 (aa 214- wt≈F or L) can be X214 or be transferred mainly to L or

F;

M42 (aa 215- wt≈T) can be X215 or be mutated mainly in Y, or in F or C;

M43 (aa 219- wt≈K) can be X219 or be transferred mainly to Q, or to E, or

N, or R, orT or W; M44 (aa 223- wt≈K) can be X223 or be mainly transferred to Q; M45 (aa 225- wt≈P) can be X225 or be transferred mainly to H; M46 (aa 227- wt≈F) can be X227 or be mainly transferred to L; M47 (aa 228- wt≈L) can be X228 or be mutated mainly in R, or in H or F;

M48 (aa 233- wt≈E) can be X233 or be mainly transferred to V, or in N M49 (aa 234- wt≈L) can be X234 or be mainly transferred to I, or in H or

P;

M50 (aa 236- wt≈P) can be X236 or be transferred mainly to M, or to S or L;

M51 (aa 238- wt≈K) can be X238 or be mainly transferred to T or S; M52 (aa 240- wt≈T) can be X240 or be transferred mainly to A, or to D or

I; at least one of M38 to M52 representing an amino acid carrying a mutation.

In a preferred embodiment of the invention,

M38 (aa 208) can be X208 or be mutated to Y;

M39 (aa 210) can be X210 or be mutated to W;

M40 (aa 211) can be X211 or mutated to K, A, E, N, D, G or Q; M41 (aa 214) can be X214 or be mutated into L or F;

M42 (aa 215) can be X215 or be mutated to Y, F or C;

M43 (aa 219) can be X219 or be mutated to Q, E N, R, T or W;

M44 (aa 223) can be X223 or be mutated to Q;

M45 (aa 225) can be X225 or be mutated to H; M46 (aa 227) can be X227 or be mutated to L;

M47 (aa 228) can be X228 or be mutated to R, H or F;

M48 (aa 233) can be X233 or be mutated to V or N;

M49 (aa 234) can be X234 or be mutated into I, H or P;

M50 (aa 236) can be X236 or be mutated to M, S or L; M51 (aa 238) can be X238 or be mutated to T or S;

M52 (aa 240) can be X240 or be mutated to A, D or I; at least one of M38, M40, M44 to M52 representing an amino acid carrying a mutation. * K bis sequence:

AA 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 D L I A E I Q K Q M53 Q G Q W T

339 340 341 342 Y Q I Y in which M53 (aa 333- wt≈G) can be mainly transferred to E or D.

The invention also relates to a pharmaceutical composition based on at least one peptide, dispersed in pharmaceutically acceptable excipients, for their use as a vaccine.

The pharmaceutical composition according to the invention may also comprise, in addition, immunomodulators such as cytokines or chemokines.

It is possible to add into the pharmaceutical composition according to the invention, adjuvants such as lipids (fatty acids, phospholipids, incomplete adjuvant of

Freund in particular), anionic copolymers, CpG units

In one embodiment of the invention, at least one peptide according to the invention is included in the composition of a lipopeptide or a lipoprotein. Indeed, the association of a peptide with a fatty acid or phospholipid sequence makes it possible to observe a potentiation of the immunogenic response compared to that brought about by the administration of the antigen alone.

The peptide (s) according to the invention can be coupled to micro or nanoparticles made up of a polysaccharide nucleus and / or covered in particular with a lipid bilayer. They can also be coupled to one or more liposomes or one or more niosomes.

The peptide (s) according to the invention can be expressed in a recombinant virus, or a recombinant viral vector, or in an attenuated or inactivated virus. The peptide (s) according to the invention can be expressed in a recombinant bacterium or in a recombinant bacterial vector.

The peptide (s) according to the invention can be expressed in an antigen presenting cell.

Another subject of the invention is a vaccine comprising at least one naked DNA or a naked RNA encoding at least one peptide according to the invention. Another object of the invention is that the agent for treating patients infected with HIV comprises at least two compositions, the first composition comprising at least one medicament from the NRTI and / or NNRTI family, and the second composition comprising at least one peptide according to the invention. These compositions are intended for a sequential application, separate or not in time or simultaneous.

Preferably, the agent for treating patients infected with HIV comprises at least two compositions, the first composition comprising at least one medicament from the NNRTI family, and the second composition comprising at least one peptide according to the invention.

Another object of the invention is a method of preventing or treating an infection with the HIV virus. This method consists in inducing a specific type T immune response in response to the administration of a vaccine according to the invention. Preferably, the mutation or mutations of the VTH virus are located in the reverse transcriptase enzyme.

Another subject of the invention is peptide sequences of 8 to 20 amino acids, recognized as epitopic sequences by T cells specific for the peptide sequences corresponding prior to vaccination, introduced to vaccination. The epitopic sequences according to the invention are peptide sequences of 8 to 20 contiguous amino acids chosen from the peptide sequences from A to K having at least one Mp mutation. Preferably, these peptide sequences consist of 8 to 10 contiguous amino acids chosen from the peptide sequences from A to K having at least one Mp mutation. More preferably, the epitopic sequences according to the invention consist of 9 contiguous amino acids chosen from the peptide sequences from A to K having at least one Mp mutation.

The epitopic sequences possessing the stated characteristics can be used for the determination and the quantification of the T cells specific for these epitopic sequences.

The subject of the invention is also a diagnostic composition for determining the T cells specific for these epitopic sequences, characterized in that it comprises, inter alia, at least one epitopic sequence according to the invention. The vaccines obtained according to the methods of the invention can be formulated as pharmaceutical compositions for subcutaneous, intramuscular, intra-dermal or intravenous administration. The techniques for administering this type of vaccine are abundantly described in the literature and are well known to those skilled in the art.

Pharmaceutically acceptable excipients can be added depending on the administration technique used. In particular, aqueous excipients such as a saline buffer solution can be added. These solutions are sterile and free from toxic or pyrogenic substances. The compositions according to the invention can be sterilized by conventional and known sterilization methods.

According to the mode of administration, the compositions according to the invention can be presented in all the forms usually used in diagnosis.

The compositions according to the invention can in particular contain pharmaceutically acceptable substances such as pH stabilizers, buffering agents or for example sodium acetate, sodium chloride, potassium, calcium, lactate sodium or others in order to be in physiological conditions.

The following examples illustrate the invention without limiting it in any way.

Example 1: Generation of CTLs lines:

The consequences of mutations induced under NRTI on specific CTL responses of RT have been studied. A total of 66 blood samples from

35 patients treated or not treated in mono- or bi-therapy with NRTI, without protease inhibitor, were collected for 1 to 6 years each year between 1991 and 1996.

To analyze the recognition of the products of the pol and rt genes of the VLH-1 virus, HIV-specific lines were generated by co-culture of PBMC (peripheral blood mononuclear cells) of patients and of autologous PHA-blasts. PBMC stimulated with PHA (phytohemagglutinin) at 2 μg / ml for 18 h at 37 ° C in a humid atmosphere at 5% CO2 and irradiated at 5000 rads are added to autologous PBMC freshly thawed at a ratio of 1: 5 and deposited in plates of 24 well at the concentration of 1x10 ^ / ml. CTLs lines are cultured for at least 3 weeks in the presence of Interleukin 2 at 20 U / ml, added on the third day of culture and then every 3 days. These lines are tested for their cytotoxic activity after at least 3 weeks of culture against autologous B-EBV lines (B lymphocytes transformed by EBV) serving as targets.

The same process was implemented with regard to the mutations induced under treatment with NNRTI or under joint treatment with NNRTI and NRTI.

Example 2: CTL test

CTL activity was measured using a chromium release test. The target cells are autologous B-EBV lines infected with the various recombinant vaccinia viras expressing the pol gene or fragments of the pol gene of HIV-1, at a multiplicity of infection of 5 PFU (Plaque Forming Unit) / cell , 18h before the CTL test. The non-recombinant wild vaccinia virus (Vac-WT) serves as a control. The target cells are incubated in a pellet in the presence of Na2 ^ Crθ4 (specific activity 2 mci / ml), at the concentration of 70 μci / xlO ^ cells, for 2 h at 37 ° C. The radioactivity measured is calculated according to the following formula: (experimental salting out - spontaneous salting out / (total salting out - spontaneous salting out) X 100 =% specific lysis. The CTL responses are considered positive when more than 10% lysis is obtained. specific, this at least 2 successive effector / target ratios.

The analysis of the recognition of the RT-1: 1-143 and RT-2: 143-293 regions was carried out on CTL lines specific for Pol. Cytotoxic T responses were assessed using a chromium 51 release test. The target cells were autologous B-EBV-lines infected with vaccinia viruses expressing different fragments of rt genes (RT-1: 1- 143 and RT-2: 143-293).

Example 3: Detection of resistance mutations:

Resistance mutations were detected in proviral DNA using a LiPA HIV-1 RT test (Murex. Diagnostics SA, Châtillon, France). This test allows the simultaneous detection of “wild” type sequences and resistance mutations (41, 69, 70, 74, 184, 214 and 215) induced by nucleoside RT inhibitors (NRTI). From the DNA isolated from PBMCs, an amplification of the rt gene is carried out using biotinylated primers, the biotinylated DNA is then hybridized with specific oligonucleotide probes immobilized in parallel lines on a strip. The addition of labeled streptavidin to alkaline phosphatase leads to its binding with the biotinylated hybrids formed which leads to the formation of a purple / brown precipitate. The wild-type and mutated sequences of rt located at the level of the different codons can be located at the level of the same strip.

Example 4: ELISpot test

For the recognition of epitopic peptides, an ELISpot-IFNγ test in which the PBMCs were incubated, without prior stimulation, with the peptide of interest was used. The 96 wells of a Immobilon p type plate (Millipore, Molsheim, France) are covered for 18 h at 4 ° C with a monoclonal anti-IFNγ (interferon γ) capture antibody (IgGl B-Bl at 1 μg / ml Diaclone, Besançon, France). After 3 washes, the

PBMCs are added in triplicate at the concentration of 1x10 ^ and 5x10 ^ cells per well in the presence of 5 μg / ml of peptide or 0.5 μg / ml of PHA, used as a positive control or of medium alone used as a negative control. The plates are then incubated for 40 h at 37 ° C. in a humid atmosphere at 5% CO 2, then washed, the biotinylated monoclonal anti-IFNγ detection antibody (B-Gl at 0.2 μg / ml Diaclone,

Besançon, France) is then added and the plates are again incubated for 4 h at 37 ° C. After washing, the streptavidin labeled with alkaline phosphatase (Amersham, Les Ulis, France) is added and the plate is incubated for 1 h at 37 ° C. The plate is then washed and then incubated with BCIP / NTP (Sigma, Aldrich, Saint Quentin, France), purple spots appear after 10 to 15 min. The plate is washed with water to stop the reaction. The spots are counted using an automatic ELISpot reader. The number of specific responder T cells is calculated after subtracting the values of the negative control. The frequency of cells secreting lTFNγ is rendered in SFC (Spot Forming Cells).

Example No. 5: Cutting the sequence B (aa 30-50) into different peptides of length varying between 8 and 21 amino acids. The peptide sequences are cut into peptides of 8 to 80 amino acids (aa) involving any part of the sequence covering at least one mutation, this for therapeutic or diagnostic applications.

For the peptide sequence 30-50 where M2 corresponds to the mutated aa 39 (A) and M3 corresponds to the mutated aa 41 (L):

aa 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 K I K A L V E I C A E L E K E G K I S K I

* Sequences of 8 aa: aa 32-39 32 33 34 35 36 37 38 39

K A L V E I C A aa 33-40 33 34 35 36 37 38 39 40

A L V E I C A E aa 34-41 34 35 36 37 38 39 40 41

L V E I C A E L aa 35-42 35 36 37 38 39 40 41 42

V E I C A E L E aa 36-43 36 37 38 39 40 41 42 43

E I C A E L E K aa 37-44 37 38 39 40 41 42 43 44

I C A E L E K E aa 38-45 38 39 40 41 42 43 44 45

C A E L E K E G aa 39-46 39 40 41 42 43 44 45 46

A E L E K E G K aa 40-47 40 41 42 43 44 45 46 47

E L E K E G K I aa 41-48 41 42 43 44 45 46 47 48

L E K E G K I S

* Sequences of 9 aa: aa 31-39 31 32 33 34 35 36 37 38 39 IKALVEICA aa 32-40 32 33 34 35 36 37 38 39 40

K A L V E I C A E aa 33-41 33 34 35 36 37 38 39 40 41

A L V E I C A E L aa 34-42 34 35 36 37 38 39 40 41 42

L V E l C A E L E aa 35-43 35 36 37 38 39 40 41 42 43

V E I C A E L E K aa 36-44 36 37 38 39 40 41 42 43 44

E I C A E L E K E aa 37-45 37 38 39 40 41 42 43 44 45

I C A E L E K E G aa 38-46 38 39 40 41 42 43 44 45 46

C A E L E K E G K aa 39-47 39 40 41 42 43 44 45 46 47

A E L E K E G K I aa 40-48 40 41 42 43 44 45 46 47 48

E L E K E G K I S aa 41-49 41 42 43 44 45 46 47 48 49

L E K E G K I S K

* Sequences of 10 aa: aa 30-39 30 31 32 33 34 35 36 37 38 39 KIKALVEICA aa 31-40 31 32 33 34 35 36 37 38 39 40 IKALVEICAE aa 32-41 32 33 34 35 36 37 38 39 40 41 KALVEICAEL aa 33-42 33 34 35 36 37 38 39 40 41 42 ALVEICAELE aa 34-43 34 35 36 37 38 39 40 41 42 43 LVEICAELEK aa 35-44 35 36 37 38 39 40 41 42 43 44 VEICAELEKE aa 36-45 36 37 38 39 40 41 42 43 44 45 E l CAELEKEG aa 37-46 37 38 39 40 41 42 43 44 45 46 ICAELEKEGK aa 38-47 38 39 40 41 42 43 44 45 46 47 CAELEKEGKI

AA 39- 39 40 41 42 43 44 45 46 47 48 48

A E L E K E G K I S aa 40-49 40 41 42 43 44 45 46 47 48 49 E L E K E G K I S K aa 41-50 41 42 43 44 45 46 47 48 49 50 L E K E G K I S K I

* Sequences of 11 aa: aa 30-40 30 31 32 33 34 35 36 37 38 39 40 KIKALVEICAE aa 31-41 31 32 33 34 35 36 37 38 39 40 41 IKALVEICAEL aa 32-42 32 33 34 35 36 37 38 39 40 41 42 KALVEICAELE aa 33-43 33 34 35 36 37 38 39 40 41 42 43 ALVEICAELEK aa 34-44 34 35 36 37 38 39 40 41 42 43 44 LVEICAELEKE aa 35-45 35 36 37 38 39 40 41 42 43 44 45 VEICAELEKEG aa 36-46 36 37 38 39 40 41 42 43 44 45 46 E l CAELEKEGK aa 37-47 37 38 39 40 41 42 43 44 45 46 47 ICAELEKEGKI aa 38-48 38 39 40 41 42 43 44 45 46 47 48 CAELEKEGKIS aa 39-49 39 40 41 42 43 44 45 46 47 48 49 AELEKEGKISK aa 40-50 40 41 42 43 44 45 46 47 48 49 50 ELEKEGKISKI * Sequences of 12 aa: aa 30-41 30 31 32 33 34 35 36 37 38 39 40 41 KIKALVEICAEL aa 31-42 31 32 33 34 35 36 37 38 39 40 41 42 IKALVEICAELE aa 32-43 32 33 34 35 36 37 38 39 40 41 42 43 KALVEICAELEK aa 33-44 33 34 35 36 37 38 39 40 41 42 43 44 ALVEICAELEKE aa 34-45 34 35 36 37 38 39 40 41 42 43 44 45 LVEICAELEKEG aa 35-46 35 36 37 38 39 40 41 42 43 44 45 46 VEICAELEKEGK aa 36-47 36 37 38 39 40 41 42 43 44 45 46 47 E l CAELEKEGKI aa 37-48 37 38 39 40 41 42 43 44 45 46 47 48 ICAELEKEGKIS aa 38-49 38 39 40 41 42 43 44 45 46 47 48 49 CAELEKEGKISK aa 39-50 39 40 41 42 43 44 45 46 47 48 49 50 AELEKEGKISKI

* Sequences of 13 aa: aa 30-42 30 31 32 33 34 35 36 37 38 39 40 41 42 KIKALVEICAELE aa 31-43 31 32 33 34 35 36 37 38 39 40 41 42 43 IKALVEICAELEK aa 32-44 32 33 34 35 36 37 38 39 40 41 42 43 44 KALVEICAELEKE aa 33-45 33 34 35 36 37 38 39 40 41 42 43 44 45 ALVEICAELEKEG aa 34-46 34 35 36 37 38 39 40 41 42 43 44 45 46 LVEICAELEKEGK aa 35-47 35 36 37 38 39 40 41 42 43 44 45 46 47 VEICAELEKEGKI aa 36-48 36 37 39 40 41 42 43 44 45 46 47 48 EIAELEKEGKIS aa 37-49 37 38 39 40 41 42 43 44 45 46 47 48 49 ICAELEKEGKISK aa 38-50 38 39 40 41 42 43 44 45 46 47 48 49 50 CAELEKEGKISKI

* Sequences of 14 aa: aa 30-43 30 31 32 33 34 35 36 37 38 39 40 41 42 43 KIKALVEICAELEK aa 31-44 31 32 33 34 35 36 37 38 39 40 41 42 43 44 IKALVEICAELEKE aa 32-45 32 33 34 35 36 37 38 39 40 41 42 43 44 45 KALVEICAELEKEG aa 33-46 33 34 35 36 37 38 39 40 41 42 43 44 45 46 ALVEICAELEKEGK aa 34-47 34 35 36 37 38 39 40 41 42 43 44 45 46 47 LVEICAELEKEGKI aa 35-48 35 36 37 38 39 40 41 42 43 44 45 46 47 48 VEICAELEKEGKIS aa 36-49 36 37 38 39 40 41 42 43 44 45 46 47 48 49 EICAELEKEGKISK aa 37-50 37 38 39 40 41 42 43 44 45 46 47 48 49 50 ICAELEKEGKISKI

* Sequences of 15 aa: aa 30-44 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 KIKALVEICAELEKE aa 31-45 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 IKALVEICAELEKEG aa 32-46 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 KALVEICAELEKEGK aa 33-47 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 ALVEICAELEKEGKI aa 34-48 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 LVEICAELEKEGKIS aa 35-49 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 VEICAELEKEGKISK aa 36-50 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 EICAELEKEGKISKI

* Sequences of 16 aa: aa 30-45 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 KIKALVEICAELEKEG aa 31-46 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 IKALVEICAELEKEGK aa 32 -47 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 KALVEICAELEKEGKI aa 33-48 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 ALVEICAELEKEGKIS aa 34-49 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 LVEICAELEKEGKISK aa 35-50 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 VEICAELEKEGKISKI

* Sequences of 17 aa: aa 30-46 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 KIKALVEICAELEKEGK aa 31-47 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 IKALVEICAELEKEGKI aa 32-48 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 KALVEICAELEKEGKIS aa33-49 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 ALVEICAELEKEGKISK aa 34-50 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 LVEICAELEKEGKISKI

* 18 aa sequences: aa 30-47 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 K l KALVEICAELEKEGKI aa 31-48 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 IKALVEICAELEKEGKIS aa 32 -49 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 KALVEICAELEKEGKISK aa 33-50 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 ALVEICAELEKEGKISKI

* Sequences of 19 aa: aa 30-48 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 KIKALVEICAELEKEGKIS aa 31-49 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 IKALVEICAELEKEGKISK aa 32-50 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 KALVEICAELEKEGKISKI

* Sequences of 20 aa: aa 30-49 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 KIKALVEICAELEKEGKISK aa 31-50 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 IKALVEICAELEKEGKISKI

* Sequences of 21 aa: aa 30-50 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 K I K A L V E I C A E L E K E G K I S K I

Example No. 6: Detection of resistance mutations on pro viral PNA isolated from blood mononuclear cells (CMNS) of patient 201 # 5.

In patient 201 # 5 treated with NRTI, the resistance mutations were detected in the proviral DNA using a LiPA HIV-1-RT test (see example 3). The LiPA test allows the definition of mutated amino acids from time to time within of a given sequence and not that of nucleic acid sequences. The method for detecting resistance mutations in DNA is summarized in Figure 1.

Using this test, we detected mutations 41, 184 and 215 in this patient. The RT sequence of patient 201 # 5 is as follows:

Xx41 (L) Xxl84 (V) Xx215 (Y or F) Xx

In which Xx represents a reference amino acid sequence.

Example No. 7: Ex-vivo evaluation of the immunogenicity of the mutation 41

(M> L) on the CMNS of Patient 201 # 5

The recognition of the mutation 41 (M> L) of RT by the CD8 cells producing IFN-gamma is studied in patient 201 # 5 treated with NRTI. The mutation 41 had been detected on the proviral DNA isolated from these same CMNS (see example 6). The recognition of wild-type and mutated peptides 33-41 was evaluated by

ELISpot.

The recognition frequency of the mutated 33-41 peptide is 95 SFC / 10 ⁶ CMNS and 3 SFC / 10 ⁶ CMNS for the wild type 33-41 peptide.

Example No. 8: Structure of a lipopeptide

The general structure of a lipopeptide is described in FIG. 2 A lipopeptide consists of a palmitic acid of formula C ₁₆ H ₃₂ O ₂ or described in the text as C ₁₅ H _3r COOH, of a lysine residue (amino acid basic added to the N-terminus of the peptide) and a peptide whose structure is chosen from the sequences described. For peptide 30-50 (sequence B):

aa 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44

K l K A L V E I C M2 E M3 E K E

45 46 47 48 49 50 G K I S K l in which M2 (aa 39) can be X39 or be mutated into A, K, N, P, M or S; M3 (aa 41) can be X41 or be mutated to L; at least one of M2 and M3 representing an amino acid carrying a mutation.

* Example of Lipopeptide constructed with the sequence 30-50 including the resistance mutations 39 and 41 induced by NRTIs

aa 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44

K l K A L V E I C A E L E K E

45 46 47 48 49 50

G K I S K I This peptide is noted in the form aa30-aa50 (aa50 = I)

In this example, the addition of lysine is done on amino acid 50 which is an isoleucine (I). The amino function of the lysine side chain is modified by a palmitic acid and the terminal amino acid C is amidated according to the following reactions: aa30-aa50-NH2 + lysine - »aa30-aa50-NH-CO-Lysine aa30- aa50 -NH-CO-Lysine + COOH-C ₁₅ H ₃₁ CONH2-aàa30-aa50 -NH-CO-Lysine-NH-CO-C ₁₅ H ₃₁

This structure varies according to industrial processes. Thus lysine can be added to the C-terminus of the peptide.

This structure is then purified and conditioned according to industrial processes meeting the conditions necessary for obtaining clinical batches before use for immunization in humans.

Claims

1. A method of treating infectious and / or tumor pathologies comprising a phase of treatment with anti-infectious and / or anti-tumor chemotherapy inducing resistance mutations and a phase of therapeutic vaccination directed against these resistance mutations.

2. Agent for treating infectious or tumor pathologies comprising at least two components:

the first component comprising a medicament intended for the treatment of infectious and / or anti-tumor pathologies inducing resistance mutations,

3. Peptides of 8 to 80 amino acids of the HIV reverse transcriptase sequence comprising at least one mutation by substitution and / or single or multiple insertion with respect to reference sequences of reverse transcriptase established in the absence of any treatment, mutation observed following treatment with NRTI and or NNRTI in patients infected with HIV, the peptides being capable of inducing a response mediated by T lymphocytes specific for this mutated sequence of HIV reverse transcriptase , with the exception of peptides of sequence Zj-YVDDZ _j , Z _r Y-IDDZ ₂ and Z _r YLDD-Z2 with Zj and Z ₂ being at least any amino acid.

4. Peptides according to claim 3, characterized in that they preferably contain from 15 to 50 amino acids.

5. Peptides according to one of claims 3 or 4, characterized in that their peptide sequence can be preceded or followed by one or more neutral amino acids to dissolve or stabilize the peptides.

6. Peptides according to any one of claims 3 to 5, of sequence or part of the following sequence comprising at least one Mp mutation: n

- with X (which will be written for the convenience of the following: Xn) being an amino acid corresponding to that described in the Los Alamos database at the same position of the peptide sequence of the VLH-1 reverse transcriptase and n being the position of an acid amino (aa) described in the Los Alamos database in the HIV-1 reverse transcriptase sequence.

- with Mp being a mutated amino acid and p being the number of the mutation observed.

* Sequence A: aa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

X X X X X Ml X X X X X X X X X in which Ml (aa 6) can be mutated into K, G or S

* Sequence B: aa 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44

X X X X X X X X X M2 X M3 X X X

45 46 47 48 49 50

X X X X X X in which M2 (aa 39) may be X39 or be mutated to A, K, N, P, M or S;

Sequence C aa 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

X X X X X Ml X X X X X X X X X X

16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

X X X X X X X X X X X X X X X

31 32 33 34 35 36 37 38 39 40 41 42 43 44 45

X X X X X X X X M2 X M3 X X X X

46 47 48 49 50 51

X X X X X X in which Ml (aa 6) can be X6 or be mutated into K, G or S;

M2 (aa 39) can be X39 or be mutated to A, K, N, P, M or S;

Sequence D

53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 XXXXXXXXX M4 XX M5 X M6 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82

M7 M8 M9 X X X M10 Mil X M12 X X X X X

83 84 85 86

X X X X in which M4 (aa 62) may be X62 or be mutated to V;

M5 (aa 65) can be X65 or be mutated to R;

M6 (aa 67) can be X67 or be mutated to N or G;

M7 (aa 68) can be X68 or be mutated to G or N;

M8 (aa 69) can be X69 or be mutated to D, N, S, A, or combined with multiple inserts like SG, SX, SSA, SSG, SSS, SEA, STS, ASG, SXX, or XXX; M9 (aa 70) can be X70 or be mutated to R, N, E or S; M10 (aa 74) can be X74 or be mutated into V or I; Ml 1 (aa 75) can be X75 or be mutated to M, I, L or T; M12 (aa 77) can be X77 or be mutated to L; at least one of M4 to M12 representing an amino acid carrying a mutation.

Sequence E

51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 aa

X X X X X X X X X X X M4 X X M5

66 67 68 69 70 71 72 73 74 75 76 77 78 79 80

X M6 M7 M8 M9 X X X M10 Mil X M12 X X X

81 82 83 84 85 86 87 88 89 90 91 92 93 94 95

X X X X X X X M13 M14 M15 X X X X X

96 97 98 99 100 101 102 103 104 105 106 107 108 109 11C

X X M16 X M17 M18 X M19 X X M20 X M21 X X

111 112 113 114 115 116 117 118 119 120 121 122 123 124 125

X X X X M22 M23 X X M24 X X X X

126 127 128

XXX in which M4 (aa 62) can be X62 or be mutated to V; M5 (aa 65) can be X65 or be mutated to R; M6 (aa 67) can be X67 or be mutated to N or G; M7 (aa 68) can be X68 or be mutated to G or N; M8 (aa 69) can be X69 or be mutated to D, N, S, A, or combined with multiple inserts like SG, SX, SSA, SSG, SSS, SEA, STS, ASG, SXX, or XXX; M9 (aa 70) can be X70 or be mutated to R, N, E or S; M10 (aa 74) can be X74 or be mutated into V or I; Ml 1 (aa 75) can be X75 or be mutated to M, I, L or T; M12 (aa 77) can be X77 or be mutated to L; M13 (aa 88) can be X88 or be mutated to C; M14 (aa 89) can be X89 or be mutated to G; M15 (aa 90) can be X90 or be mutated to I; M16 (aa 98) can be X98 or be mutated into G or S; M17 (aa 100) can be X100 or be mutated to I; M18 (aa 101) can be X101 or be mutated to E, Q, R, I or P; M19 (aa 103) can be X103 or be mutated to N, Q, R, T or S; M20 (aa 106) can be X106 or be mutated to A, I or L; M21 (aa 108) can be X108 or be mutated to I; M22 (aa 115) can be XI 15 or be mutated to F; M23 (aa 116) can be XI 16 or be mutated to Y; M24 (aa 119) can be XI 19 or be mutated to S; at least one of M4 to M24 representing an amino acid carrying a mutation.

Sequence F aa 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 X X X X X X X X X M25 M26 X M27 X X

144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 X X X X X X X M28 X X X X X M29 X

159 160 161 162 163 164 165 166 X X X X X X X X in which M25 (aa 138) can be X138 or be mutated into A, G, K or Q;

M26 (aa 139) can be X139 or be mutated to I, M or K;

M27 (aa 141) can be X141 or be mutated to E;

M28 (aa 151) can be X151 or be mutated to M;

M29 (aa 157) can be X157 or be mutated to S; at least one of M25 to M29 representing an amino acid carrying a mutation.

Sequence G 1 63 164 165 166 167 168 169 170 171 172 173 174 175 176 177 aa

X X X X X X X X X M30 X X X X X

178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 M31 M32 X M33 X X M34 X X X M35 M35 M37 X X

M31 (aa 178) can be X178 or be mutated to M, L or F;

M32 (aa 179) can be X179 or be mutated into I, L, D or E;

M33 (aa 181) can be X181 or be mutated to C or I;

M34 (aa 184) can be XI 84 or be mutated to V, I or T;

M35 (aa 188) can be X188 or be mutated into L, C or H;

M36 (aa 189) can be XI 89 or be mutated to I;

M37 (aa 190) can be X190 or be mutated to A, S, E, Q or T; at least one of M30 to M37 representing an amino acid carrying a mutation.

H sequence

169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 aa

X X X M30 X X X X X M31 M32 X M33 X X

184 185 M34 X in which M30 (aa 172) may be X172 or be mutated to K;

M31 (aa 178) may be X178 or be mutated to M, L or F;

M32 (aa 179) can be X179 or be mutated into I, L, D or E;

M33 (aa 181) can be X181 or be mutated to C or I;

M34 (aa 184) can be XI 84 or be mutated to V, I or T; at least one of M30 to M34 representing an amino acid carrying a mutation.

Sequence I

aa 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 M34 X X X M35 M36 M37 X X X X X X X X X in which M34 (aal84) can be XI 84

M35 (aa 188) can be X188 or be mutated into L, C or H; M36 (aa 189) can be XI 89 or be mutated to I;

Sequence J aa 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 X X X X X X X X X M38 X M39 M40 X X

214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 M41 M42 X X X M43 X X X M44 X M45 X M46 M47

229 230 23 1 232 233 234 235 236 237 238 239 240 241 242 243 X X X X M48 M49 X M50 X M51 X M52 X X X

M39 (aa 210) can be X210 or be mutated to W;

M40 (aa 211) can be X211 or mutated to K, A, E, N, D, G or Q;

M41 (aa 214) can be X214 or be mutated into L or F;

M42 (aa 215) can be X215 or be mutated to Y, F or C;

M43 (aa 219) can be X219 or be mutated to Q, E N, R, T or W;

M44 (aa 223) can be X223 or be mutated to Q;

M45 (aa 225) can be X225 or be mutated to H;

M46 (aa 227) can be X227 or be mutated to L;

M47 (aa 228) can be X228 or be mutated to R, H or F;

M48 (aa 233) can be X233 or be mutated to V or N;

M49 (aa 234) can be X234 or be mutated into I, H or P;

M50 (aa 236) can be X236 or be mutated to M, S or L;

M51 (aa 238) can be X238 or be mutated to T or S;

M52 (aa 240) can be X240 or be mutated to A, D or I; at least one of M38 to M52 representing an amino acid carrying a mutation.

Sequence K aa 324 325 326 327 328 329 330 33 1 332 333 334 335 336 337 338

X X X X X X X X X M53 X X X X X

339 340 341 342 XXXX wherein M53 (aa 333) can be mutated to E or D;

7. Pharmaceutical composition, characterized in that it comprises at least one peptide according to claims 3 to 6, dispersed in pharmaceutically acceptable excipients.

8. Peptides according to one of claims 3 to 6 or composition according to claim 5, for their use as a vaccine.

9. Vaccine characterized in that it contains at least one peptide as defined in any one of claims 3 to 6.

10. Vaccine characterized in that it contains at least one peptide as defined in any one of claims 3 to 6 to which are added immunomodulators such as cytokines or chemokines.

11. Vaccine characterized in that it contains at least one peptide as defined in any one of claims 3 to 6 to which are added adjuvants such as lipids (fatty acids, phospholipids, incomplete Freund's adjuvant), anionic copolymers, CpG units

12. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is coupled to a lipopeptide.

13. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is coupled to a lipoprotein.

14. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is coupled to micro or nanoparticles (to be defined).

15. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is coupled to one or more liposomes.

16. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is coupled to one or more niosomes.

17. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is expressed in a recombinant virus.

18. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is expressed in an attenuated or inactivated virus.

19. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is expressed in a recombinant viral vector.

20. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is expressed in a recombinant bacterium.

21. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is expressed in a recombinant bacterial vector.

22. Vaccine according to one of claims 9 to 11, characterized in that at least one peptide according to claims 3 to 6 is expressed in an antigen presenting cell.

23. Vaccine according to one of claims 9 to 11, characterized in that it comprises at least one naked DNA coding for at least one peptide as described in any one of claims 3 to 6.

24. Agent for treating patients infected with HIV, comprising at least two compositions, the first composition comprising at least one medicament from the NRTI and / or NNRTI family, and the second composition comprising at least one peptide as defined in 'any one of claims 3 to 6, these compositions being intended for a sequential application, separate or not in time or simultaneous.

25. A method of preventing or treating an infection with the HIV virus consisting in inducing a specific type T immune response in response to the administration of a vaccine as described in any one of claims 9 at 23.

26. The method of claim 25, characterized in that the mutation or mutations of the VLH virus are located in the enzyme reverse transcriptase.

27. Peptide sequences of 8 to 20 amino acids, characterized in that they are recognized as epitopic sequences by the T cells specific for the peptide vaccine sequences introduced for vaccination.

28. Epitopic sequences according to claim 27, characterized in that they are peptide sequences of 8 to 20 contiguous amino acids chosen from the peptide sequences from A to K having at least one Mp mutation.

29. Epitopic sequences according to claim 27, characterized in that they are peptide sequences of 8 to 10 contiguous amino acids chosen from the peptide sequences from A to K having at least one Mp mutation.

30. Epitopic sequences according to claim 27, characterized in that they are peptide sequences of 9 contiguous amino acids chosen from the peptide sequences from A to K having at least one Mp mutation

3 1. Use of the epitopic sequences according to any one of claims 27 to 30, for the determination and quantification of the T cells specific for these epitopic sequences.

32. A diagnostic composition for determining the T cells specific for these epitopic sequences, characterized in that it comprises at least one epitopic sequence as defined in any one of claims 27 to 30.