MULTI-EPITOPIC PEPTIDE COMPOUNDS AND VACCINES AGAINST LEISHMANIASIS
20210213114 · 2021-07-15
Assignee
Inventors
- Rachel Bras Gonçalves (Montpellier, FR)
- Jean-Loup Lemesre (Montpellier, FR)
- Amel Garnaoui (Tunis, TN)
- Elodie Petitdidier-Lesin (Prades-le-Lez, FR)
Cpc classification
Y02A50/30
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
A61K2039/55555
HUMAN NECESSITIES
International classification
Abstract
The invention relates to multi-epitopic peptide compounds obtained from PSA, HwB and LmLRAB proteins of Leishmania as well as to pharmaceutical compositions and vaccines for use against one or more leishmaniases.
Claims
1. Peptidic compound comprising at least two epitopes contained in the protein sequences chosen from the Leishmania PSA, H2B or LmLRAB proteins having a sequence chosen from the group consisting of sequences SEQ ID NO: 1 to 21, sequences SEQ ID NO: 24 to 64, sequences SEQ ID NO: 67, 68, 70 to 72, 74 to 76, sequences SEQ ID NO: 78 to 95, sequences SEQ ID NO: 98 to 136, sequences SEQ ID NO: 139 to 156, sequences SEQ ID NO: 158 to 184, and analogous, mutein and homologous derivatives thereof, the epitopes optionally being separated by a peptide spacer comprising at least one amino acid.
2. Peptidic compound according to claim 1, wherein the analogous, mutein and homologous derivatives of the epitopes, having immunogenic capability, have a percentage identity of sequence of at least 50%, preferably at least 75% with respective sequences of the epitopes.
3. Peptidic compound according to claim 1 wherein the peptidic spacer comprises 1 to 8 amino acids.
4. Peptidic compound according to claim 1, which comprises 2, 3 or 4 of the epitopes.
5. Peptidic compound according to claim 1, wherein a first epitope is chosen from the group consisting of epitopes contained in a sequence of a first protein chosen from the group consisting of PSA, H2B and LmLRAB, and a second epitope is chosen from the group consisting of epitopes contained in a sequence of a second protein, different from the first protein, chosen from the group consisting of PSA, H2B and LmLRAB.
6. Peptidic compound according to claim 1, which is linked to a carrier group that increases the immunogenic character thereof.
7. Peptidic compound according to claim 1, wherein the epitope or epitopes is/are chosen from the group consisting of epitopes having sequences SEQ ID NO: 22, 23, 65, 66, 69, 73, 77, 96, 97, 137, 138, 157 and 185.
8. Peptidic compound according to claim 1, which includes at least three epitopes contained in the sequences of different proteins chosen from the group consisting of PSA, H2B and LmLRAB.
9. Specific antibody and immunoserum containing same, directed against epitopes of the peptidic compounds according to claim 1.
10. Pharmaceutical composition comprising at least one peptidic compound according to claim 1.
11. Composition comprising at least one peptidic compound according claim 1, wherein the composition is formulated for use in prophylactic and therapeutic vaccination directed against Leishmania.
12. Composition for use thereof according to claim 11, wherein the Leishmania are is selected from the group consisting of Leishmania donovani, Leishmania infantum, Leishmania chagasi, Leishmania mexicana, Leishmania amazonensis, Leishmania venezuelensis, Leishmania tropica, Leishmania major, Leishmania aethiopica, Leishmania (Viannia) braziliensis, Leishmania (Viannia) guyanensis, Leishmania (Viannia) panamensis, Leishmania (Viannia) peruviana, and combinations of two or more thereof.
13. Composition comprising at least one peptidic compound according to claim 1, wherein the composition is formulated for manufacturing a drug or a vaccine an in vivo or in vitro diagnostic reagent for inducing or diagnosing in mammals an activation of type Th1 lymphocyte-dependent cell-mediated immunity and/or effector humoral immunity.
14. Prophylactic and/or therapeutic vaccine for use thereof against one or more of the Leishmania chosen from Leishmania donovani, Leishmania infantum, Leishmania chagasi, Leishmania mexicana, Leishmania amazonensis, Leishmania venezuelensis, Leishmania tropica, Leishmania major, Leishmania aethiopica, Leishmania (Viannia) braziliensis, Leishmania (Viannia) guyanensis, Leishmania (Viannia) panamensis, and/or Leishmania (Viannia) peruviana, comprising at least one peptidic compound according to claim 1.
15. Vaccine according to claim 14, which comprises at least one peptidic compound chosen from the group consisting of SEQ ID NO: 22, 23, 69, 73, 77, 65, 66, 96, 97, 137, 138, 157, 185, and analogous, mutein and homologous derivatives thereof.
16. Vaccine according to claim 14, which comprises, firstly, at least one peptidic compound chosen from the group consisting of SEQ ID NO: 22, 23, 69, 73, 77, and analogous, mutein and homologous derivatives thereof; and secondly, at least one peptide compound chosen from the group consisting of sequences SEQ ID NO: 65, 66, 96, 97, 137, 138, 157, 185, and analogous, mutein and homologous derivatives thereof.
17. Vaccine according to claim 14, which comprises, in combination, the following multiepitopic peptidic compounds: each of SEQ ID NO: 23, 66, 73, 77, 97, 138, 157 and 185 or analogous, mutein and homologous derivatives thereof; or each of SEQ ID NO: 23, 66, 157 and 185 or analogous, mutein and homologous derivatives thereof; or each of SEQ ID NO: 66, 97, 138, 157 and 185 or analogous, mutein and homologous derivatives thereof; or each of SEQ ID NO: 66, 157 and 185 or analogous, mutein and homologous derivatives thereof.
18. Vaccinei according to claim 14, which further comprises an adjuvant chosen from the group consisting of adjuvants in classes TLR3, TLR4, TLRS, TLR7, TLR8, TLR9, saponins, QA21, quilA and QS21 derivatives thereof, oil in water or emulsions. water in oil emulsions, polysaccharides, cationic liposomes, virosomes, polyelectrolytes, and immunomodulators chosen from the group consisting of sandfly saliva proteins, cytokines, peptides and heat shock proteins.
19. Vaccine according to claim 14, which is formulated for at least one of administration selected from the group consisting of subcutaneous, intradermal, intramuscular, parenteral, endonasal, mucosal and oral administration.
20. Nucleotide sequence coding for an epitope contained in the protein sequences chosen from the Leishmania PSA, H2B or LmLRAB proteins having a sequence chosen from the group consisting of sequences SEQ ID NO: 1 to 21, sequences SEQ ID NO: 24 to 64, sequences SEQ NO: 67, 68, 70 to 72, 74 to 76, sequences SEQ ID NO: 78 to 95, sequences SEQ ID NO: 98 to 136, sequences SEQ ID NO: 139 to 156, sequences SEQ ID NO: 158 to 184, and analogous, mutein and homologous derivatives thereof, the epitopes optionally being separated by a peptide spacer comprising at least one amino acid.
21. Expression vector comprising at least one nucleotide sequence according to claim 20, and equipment necessary for expression thereof.
22. Diagnostic reagent comprising a peptidic compound according to claim 1.
Description
BRIEF DESCRIPTION OF THE FIGURES
[0021] The invention will be better understood from a reading of the following non-limitative description, drawn up with regard to the accompanying drawings, wherein:
[0022]
[0023]
[0024]
[0025]
DETAILED DESCRIPTION OF THE INVENTION
[0026] According to the invention, epitope means a peptide compound or a peptide defined by the sequence thereof having approximately 8 to 15 amino acids.
[0027] According to the invention, analogous derivatives or mutein derivatives of a peptide compound means the biologically active derivatives of the reference molecules that have the required activity, namely the ability to stimulate a cell-mediated immune response. In general, the term analogous derivatives refers to compounds having a sequence and a polypeptide structure having one or more additions, substitutions and/or deletions of amino acids, with respect to the peptide compounds defined above, insofar as these modifications do not destroy the immunogenic activity. According to the invention, the analogs particularly preferred include the preserving substitutions, that is to say the substitutions or replacements without any consequences on the function and the final structure of the protein. The term mutein derivative means the peptides having one or more elements imitating the peptide. Methods for preparing conventional analogs and muteins are known to persons skilled in the art.
[0028] According to the invention, homologous derivatives means peptide compounds having a certain percentage of peptide identify. The term identity signifies that the amino acids of two peptide sequences compared correspond exactly. The percentage of identity is determined by a direct comparison of the sequences between two peptide compounds by aligning said sequences and counting the exact number of mismatches between the two aligned sequences. Next, a division by the length of the shortest sequence is carried out and the result is multiplied by a hundred. The percentage identity can also be determined by means of computer programs well known to persons skilled in the art. Thus, according to the invention, two peptide sequences are said to be substantially homologous with respect to each other provided that they have at least 50%, preferably at least 70%, preferably again at least 75%, preferably again at least 85%, preferably again at least 90% and even more preferably at least 95% or more identity of sequence over a defined length of the peptide molecules.
[0029] The vaccine strategy according to the invention is intended to respond to the need for a vaccine providing good coverage of the worldwide human population and protecting against the main species of leishmaniasis. It is based on fragments of antigenic peptides capable of lastingly activating specific cell immunity directed against these parasites. Peptide vaccination is based on the molecular and cellular bases of the recognition of the antigen by the T cells. The establishment of specific immunity depends, to a large extent, on the degradation and the association of the antigenic fragments, peptides, with the molecules of the major histocompatibility complex (MHC; HLA for humans). This association is made specific of a particular HLA molecule by amino acid residues constituting the anchor units of the peptide. The complexes thus formed are recognized by the T lymphocytes by means of a membrane receptor (TcR) and requires a specific interaction with certain amino acids of the T epitope. The T epitopes are ligands of the HLA molecules with strong or moderate affinities. They are presented to the CD8+ (cytotoxic) or CD4+ (auxiliary) T lymphocytes by the HLA molecules respectively of class I or class II. The formation of these tri-molecular complexes (TcR/HLA/peptide) is the prerequisite for the activation and the expansion of the specific T cells and therefore for the induction of a protective immune response during an infection.
[0030] The vaccine strategy according to the invention relies on the identification and selection of immunodominant peptides carried by the sequences of the following specific proteins of Leishmania: [0031] i) a virulence protein characterized as the major immunogen of the antigens excreted/secreted by Leishmania, namely the soluble PSA protein, which is common and very much preserved within the species of leishmania, responsible for the various human infections;
[0032] ii) an H2B protein, which is highly preserved among the species of Leishmania, which is capable of inducing protection in the murine model and a cell response in humans, and the N-terminal region of which, which is the most divergent with respect to the histone proteins of mammals, is capable of conferring significant protection against a challenge with virulent parasites in BALB/c mice. The H2B protein is recognized by T lymphocytes of individuals cured of L. tropica or L. major, with an induction of high levels of IFN-; and [0033] iii) an LmLRAB protein, belonging to the superfamily of RAB GTPases, and highly preserved with other species of Leishmania, capable of inducing significantly high levels of IFN- in individuals cured of cutaneous leishmaniasis.
[0034] 15
[0035] The advantage of such peptides is multiple: [0036] (i) they have high innocuity since they avoid any infectious risk related to the use of a pathogenic agent or infectious agent; [0037] (ii) they are chemically well defined and thus meet the pharmaceutical requirements for purity; [0038] (iii) they facilitate monitoring of the induced immune response (search for cytotoxic T lymphocytes (CTLs) directed against a defined T epitope); and [0039] (iv) their sequence can be modified in order to make them more immunogenic.
[0040] Such peptides meet in a remarkable fashion all the conditions mentioned in the preamble of the present application: reproducible vaccine, thermostable facilitating storage and transport thereof, multipurpose, easy to produce at low cost in the endemic zones, making it possible to use it on a large scale.
[0041] Advantageously, the epitopes or peptide compounds that are the object of the invention are linked to carriers for making them more immunogenic. By way of non-limitative examples of carriers, mention can be made of the KLM (keyhole limpet hemocyanin) carrier proteins, and the lipopeptides of the palmitoyl type, or derivatives thereof. The most usual modifications to proteins by lipids are: isoprenylation, N-myristoylation, palmitoylation (or S-acylation) and glypiation. Isoprenylation and N-myristoylation are co-translational or immediately post-translational modifications and the group that is attached remains so until the protein degrades. Palmitoylation is post-translational. This modification is reversible and quicker than the turnover or degradation of the proteins:
[0042] it can therefore be regulated. Glypiation is co- and post-translational. Palmitoylated peptides are particularly advantageous according to the invention since these derivatives interact with the lipidic components of the membrane of the target cells (macrophages, dendritic cells, neutrophils, etc.), assist penetration thereof and convey them inside them in order then to present them to the immune system.
[0043] The epitopes according to the invention furthermore advantageously have one or more protective groups. This is because, so as to improve resistance to degradation, it may be opportune to use a protected form of the peptide according to the invention. The protection form is a biologically compatible form and is compatible with use in the pharmaceutical field. Numerous forms of biologically compatible protection can be envisaged, such as for example acylation or acetylation of the amino-terminal end, or amidation or esterification of the carboxy-terminal end, as is the case, for example, with compounds obtained by chemical synthesis with the addition of a palmitoylated tail at the amino-terminal end and amidation of the carboxy-terminal end, with levels of IFN- secreted by the PBMCs of individuals cured of cutaneous leishmaniasis and stimulated in vitro by mixtures of peptide compounds of the present invention in accordance with a short 10-day protocol with stimulation at D0 and addition of recombinant IL-2 at D1, D4 and D7. The levels of IFN- obtained with a combination of the peptide compounds according to the invention also shows a high added value of the compounds according to the invention. Thus the invention also relates to an epitope as defined previously, characterized by the fact that it is in protected form. It is possible to use a protection based on a substitution on the amino-terminal end by an acetyl group, a benzoyl group, a tosyl group or a benzyloxycarbonyl group. Preferably, use is made of a protection based on the amidation of the hydroxyl function of the carboxy-terminal end by an NYY group with Y representing a C1 to C4 alkyl chain, or esterification by an alkyl group. It is possible to protect both ends of the peptide compound.
[0044] The peptide derivatives according to the invention also relate to the amino acids and peptides linked together by a pseudopeptide bond. Pseudopeptide bond means all the types of bond able to replace conventional peptide bonds. In the field of amino acids, the geometry of the molecules is such that they can theoretically be in the form of different optical isomers. There exists, in fact, a molecular conformation of amino acid (aa) such that it diverts to the right the polarization plane of light (dextrorotatory or D-aa conformation), and a molecular conformation of the amino acid (aa) such that it diverts to the left the polarization plane of light (levogyre or L-aa conformation). Natural amino acids are always of levogyre conformation, and consequently a peptide of natural origin will consist only of amino acids of the L-aa type. However, chemical synthesis in the laboratory makes it possible to prepare amino acids having both possible conformations.
[0045] From this basic material, it is thus possible, when synthesizing a peptide, to incorporate amino acids in the form of both dextrorotatory or levogyre optical isomers. Thus the amino acids constituting the peptide according to the invention may be in L-, D- or DL-configuration.
[0046] The epitopes and peptide compounds according to the invention can be obtained either by conventional chemical synthesis in solid phase or in liquid homogeneous phase, or by enzymatic synthesis, from constituent amino acids or derivatives thereof. The epitopes and peptide compounds according to the invention can also be obtained by fermentation of a strain of bacteria, modified or not, by genetic engineering, or by extraction of proteins of animal or vegetable origin, preferably of vegetable origin, followed by a controlled hydrolysis that releases peptidic fragments corresponding totally or partially to the epitopes and peptide compounds according to the invention.
[0047] The Applicant has been able to show that the various epitopes according to the invention are consensus sequences common to the main species of Leishmania and have a strong or moderate affinity for all the molecules of the MHC (major histocompatibility complex) of mammals, and more particular for all the molecules of the HLAs (HLAs standing for human leucocyte antigens), mainly represented by the human populations most seriously affected by these ailments.
[0048] This is because, in the context of a strategy of vaccination of human populations exposed to Leishmania infections and in order to meet the need for a vaccine providing good coverage of the world population and protecting against leishmaniases, it is important to use antigen/peptide fragments capable of activating, lastingly, specific cell immunity directed against the parasite.
[0049] In order to ensure good coverage of the world population and having regard to the great variability of the HLA phenotype (the major human histocompatibility complex) between individuals, the immunogenic antigen fragments (peptides) of sufficient length must contain a series of epitopes able to be presented by several types of HLA molecules of class I and II.
[0050] HLA molecules are highly polymorphous. This is because there exist more than 2500 HLA proteins of class I (HLA-I) and more than 1000 HLA proteins of class II (HLA-II). However, some of these HLA molecules, close in sequence and in spatial conformation, may have epitopes common to the T cells. The grouping of several thousands of HLA molecules is at the present time described in a little more than 20 or so categories, referred to as HLA supertypes having epitopes that are very well preserved for each supertype.
[0051] To the development of peptide vaccines, there is added the multiepitope or polyepitope approach (a peptide containing a plurality of epitopes). This multiepitope approach is advantageous for developing a vaccine intended for the whole of the world population. This is because, in order to ensure good coverage of the world population and having regard to the great variability of the HLA phenotype between individuals, immunogenic antigen fragments (peptides) of sufficient length must contain a series of epitopes able to be presented by several supertypes of HLA-I and -II molecules.
[0052] Thus the epitopes included in the peptide compounds of the invention have great immunogenic capacity.
[0053] T epitopes are antigen sequences that recognize T lymphocytes. For example, in humans, T epitopes result from the degradation of antigens by the presenting cells and are presented to the CD8+ (cytotoxic) or CD4+ (auxiliary) T lymphocytes by the HLA molecules respectively of class I or class II. T epitopes are therefore necessarily ligands of HLA molecules and effectively form part of the peptides, which bind to the HLA molecules with strong and moderate affinities.
[0054] The cells expressing the major histocompatibility complex of class II (CMH2) may also present microbial antigens via CD1 to gamma-delta T lymphocytes.
[0055] The presentation capabilities depend on numerous variables. They vary from one individual to another because of the polymorphism of the major histocompatibility complex (MHC).
[0056] Thus consanguinity, by reducing the number of different MHCs expressed by an individual, reduces their immune capacities. They also differ according to the ways of exposure to the antigen (dose and administration route), because of the variations in the presentation capacities of the various types of presenting cells. For example, the cells involved in the presentation will be different by cutaneous or digestive route.
[0057] Finally, the peptide range produced by a given antigen will be different according to the presenting cell (cleavage methods), and according to the species and the individual (the allele of the MHC).
[0058] The peptide compounds according to the invention have been selected and designed so as to ensure vaccinal and therapeutic coverage of the populations most seriously affected by the main pathogenic species of Leishmania. They are intended to induce and to characterize the prevention or treatment of ailments in mammals the protective immunity of which depends on the stimulation of the type Th1 leucocytes and cytotoxic T cells, characteristic of a state of hyperstimulation of a delayed type.
[0059] As is described above, another main difficulty in the development of a vaccine candidate lies in the fact that it must ideally be effective against several species of Leishmania and in particular against the most severe clinical forms (visceral and cutaneous) and in various natural hosts of the infection (humans, dogs).
[0060] The sequences of these epitopes are preferentially: [0061] the peptide sequence of H2B proteins contained in Table 1 below; [0062] the peptide sequences of LmLRAB proteins contained in Table 2 below; [0063] the peptide sequences of PSA proteins contained in Table 3 below,
as well as the analogous, mutein and homologous derivatives thereof.
TABLE-US-00002 TABLE1 H2B Affinityforthe Affinityforthe Affinityforthe HLAallelesof HLAsupertypes HLAallelesof SEQIDH2B-1 Peptide classI ofclassI classII SEQIDNO:1 AINAQMSMM HLA-B*15:01 HLA-B*62 SEQIDNO:2 IKAINAQMSM HLA-B*15:01, HLA-B*58, HLA-B*57:01 HLA-B*62 SEQIDNO:3 KAINAQMSM HLA-B*58:01, HLA-A*01/HLA- HLA- HLA-B*57:01, A*03, DRB4*01:01 HLA-B*15:01, HLA-B*58, HLA-A*30:01 HLA-B*62 SEQIDNO:4 KAINAQMSMM HLA-B*58:01, HLA-B*58, HLA-B*57:01, HLA-B*62 HLA-B*15:01 SEQIDNO:6 MERICTEAA HLA-B*40:02, HLA-B*44 HLA-B*45 :01 SEQIDNO:5 MMERICTEA HLA-A*02:03, HLA-A*02 HLA-A*02:01 SEQIDNO:7 MMERICTEAA HLA-A*02:03 HLA-A*02 SEQIDNO:8 MSHRTMKIK HLA-A*30:01, HLA-A*01/HLA-A*03, HLA-A*03:01, HLA-A*11:01 HLA-A*03 SEQIDNO:9 MSHRTMKIKA HLA-A*30:01 HLA-A*01/HLA-A*03 SEQIDNO:10 MSHRTMKSM HLA-B*57:01, HLA-A*01/HLA-A*03, HLA-A*30:01, HLA-B*15:01 HLA-B*58, HLA-B*62 SEQIDNO:11 MSMSHRTM HLA-B*08:01 HLA-B*08 SEQIDNO:12 MSMSHRTMK HLA-A*11:01, HLA-A*01/HLA- HLA- HLA-A*31:01, A*03,HLA-A*03 DRB1*03:01, HLA-A*03:01, HLA- HLA-A*68:01, DRB1*09:01, HLA-A*30:01, HLA- HLA-A*33:01 DRB1*11:01 SEQIDNO:13 MSMSHRTMKS HLA-A*03:01, HLA-A*03 HLA-A*11:01 SEQIDNO:14 NAQMSMMER HLA-A*68:01, HLA-A*03 HLA-A*33:01, HLA-A*31:01 SEQIDNO:15 SMSHRTMK HLA-A*03:01 HLA-A*03 SEQIDNO:16 SMSHRTMKI HLA-A*02:03, HLA-A*02 HLA- HLA-A*32:01, DRB1*07:01, HLA-A*02:01 HLA- DRB1*13:02 SEQIDNO:17 SMSHRTMKIK HLA-A*03:01, HLA-A*01/HLA- HLA-A*11:01, A*03,HLA-A*03 HLA-A*30:01 SEQIDNO:18 SMSHRTMKS HLA-A*02:03 HLA-A*02 SEQIDNO:19 SMSHRTMKSM HLA-B*15:01, HLA-A*02, HLA- HLA-B*57:01, B*08, HLA-B*58, HLA-A*02:03, HLA-B*62 HLA-B*08:01 SEQIDNO:20 TMKSMSHRT HLA-A*02:03 HLA-A*02 SEQIDNO:21 TMKSMSHRTM HLA-B*15:01, HLA-B*08, HLA- HLA-B*08:01 B*62 SEQIDNO:22 MSMSHRTMKSMSHRTMKIKAINAQMSMMERICTEAA SEQIDNO:23 KARYMSMSHRTMKSMSHRTMKIKAINAQMSMMERICTEAA SEQID Concatenationaffinity H2B-II Peptide fortheHLAallelesofclassII SEQIDNO:24 RKPKRSWNVYVGRSL HLA-DRB1*13:02,HLA-DRB1*01:01,HLA-DRB1*07:01, HLA-DRB1*09:01,HLA-DRB1*15:01 SEQIDNO:25 KPKRSWNVYVGRSLK HLA-DRB5*01:01,HLA-DRB1*01:01,HLA-DRB1*07:01, HLA-DRB1*09:01,HLA-DRB1*15:01 SEQIDNO:26 PKRSWNVYVGRSLKA HLA-DRB5*01:01,HLA-DRB1*01:01,HLA-DRB1*11:01, HLA-DRB5*13:02,HLA-DRB5*07:01,HLA-DRB5*09:01, HLA-DRB5*15:01 SEQIDNO:27 KRSWNVYVGRSLKAI HLA-DRB1*01:01,HLA-DRB1*03:01,HLA-DRB1*04:01, HLA-DRB1*11:01,HLA-DRB1*13:02,HLA-DRB1*07:01, HLA-DRB1*09:01,HLA-DRB1*15:01,HLA-DRB5*01:01, HLA-DPA1*02:01/DPB1*01:01 SEQIDNO:28 RSWNVYVGRSLKAIN HLA-DRB5*01:01,HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*11:01,HLA-DRB1*13:02, HLA-DRB1*04:05,HLA-DRB1*07:01,HLA-DRB1*09:01, HLA-DRB1*15:01,HLA-DPA1*02:01/DPB1*01:01, HLA-DRB1*08:02 SEQIDNO:29 SWNVYVGRSLKAINA HLA-DRB5*01:01,HLA-DRB1*04:05,HLA-DPA1*03:01/ DPB1*04:02,HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*11:01,HLA-DRB1*13:02, HLA-DRB1*07:01,HLA-DRB1*09:01,HLA-DRB1*15:01, HLA-DPA1*02:01/DPB1*01:01,HLA-DRB1*08:02 SEQIDNO:30 WNVYVGRSLKAINAQ HLA-DRB5*01:01,HLA-DRB1*04:05,HLA-DPA1*03:01/ DPB1*04:02,HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*11:01,HLA-DRB1*07:01, HLA-DRB9*01:01,HLA-DRB1*15:01,HLA-DPA1*02:01/ DPB1*01:01,HLA-DRB1*08:02 SEQIDNO:31 NVYVGRSLKAINAQM HLA-DRB1*04:01,HLA-DRB4*01:01,HLA-DRB1*03:01, HLA-DRB1*04:05,HLA-DRB1*11:01,HLA-DRB1*01:01, HLA-DRB1*09:01,HLA-DRB1*15:01,HLA-DRB1*08:02 SEQIDNO:32 VYVGRSLKAINAQMS HLA-DRB1*01:01,HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB4*01:01,HLA-DRB1*04:01,HLA-DRB1*09:01, HLA-DRB1*03:01,HLA-DRB1*11:01,HLA-DRB1*15:01, HLA-DRB1*08:02 SEQIDNO:33 YVGRSLKAINAQMSM HLA-DRB1*01:01,HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*09:01,HLA-DRB4*01:01,HLA-DRB5*01:01, HLA-DRB1*04:01,HLA-DRB1*11:01,HLA-DRB1*08:02, SEQIDNO:34 VGRSLKAINAWMSMS HLA-DRB1*01:01,HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*09:01,HLA-DRB4*01:01,HLA-DRB5*01:01, HLA-DRB1*04:01,HLA-DRB1*08:02,HLA-DRB1*11:01, HLA-DRB1*13:02 SEQIDNO:35 GRSLKAINAQMSMSH HLA-DRB1*15:01,HLA-DRB1*01:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*08:02,HLA-DRB1*09:01, HLA-DRB1*11:01,HLA-DRB4*01:01,HLA-DRB5*01:01, HLA-DRB1*04:01,HLA-DRB1*13:02 SEQIDNO:36 RSLKAINAWMSMSHR HLA-DRB1*04:01,HLA-DRB1*15:01,HLA-DRB1*05:01, HLA-DRB1*01:01,HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*08:02,HLA-DRB1*09:01,HLA-DRB1*11:01, HLA-DRB4*01:01, SEQIDNO:37 SLKAINAQMSMSHRT HLA-DRB1*07:01,HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*09:01,HLA-DRB1*11:01,HLA-DRB1*15:01, HLA-DRB5*01:01,HLA-DRB1*01:01,HLA-DRB1*08:02, HLA-DRB4*01:01, SEQIDNO:38 LKAINAQMSMSHRTM HLA-DRB1*07:01,HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*09:01,HLA-DRB1*11:01,HLA-DRB1*15:01, HLA-DRB5*01:01,HLA-DRB1*01:01,HLA-DRB4*01:01, SEQIDNO:39 KAINAQMSMSHRTMK HLA-DRB1*07:01,HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*09:01,HLA-DRB1*11:01,HLA-DRB1*15:01, HLA-DRB5*01:01,HLA-DRB4*01:01,HLA-DRB1*03:01, HLA-DRB1*01:01 SEQIDNO:40 AINAQMSMSHRTMKI HLA-DRB1*01:01,HLA-DRB1*04:01,HLA-DRB1*09:01, HLA-DRB1*15:01,HLA-DRB4*01:01,HLA-DRB5*01:01, HLA-DRB3*01:01,HLA-DRB1*11:01,HLA-DRB1*07:01 SEQIDNO:41 INAQMSMSHRTMKIV HLA-DRB1*04:01,HLA-DRB1*15:01,HLA-DRB1*05:01, HLA-DRB1*03:01,HLA-DRB1*09:01,HLA-DRB1*11:01, HLA-DRB1*01:01,HLA-DRB4*01:01,HLA-DRB1*07:01, HLA-DRB1*13:02 AffinityHLA AffinityHLA AffinityHLA Peptide- allelesof allelesof supertypeallele coreepitope classII classI ofclassI SEQIDNO:42 AQMSMSHRT HLA-DRB1*01:01, HLA-DRB1*07:01 SEQIDNO:43 GRSLKAINA HLA-DRB1*01:01 SEQIDNO:44 INAQMSMSH HLA-DRB1*04:01, HLA-DRB1*04:05, HLA-DRB1*09:01, HLA-DRB1*11:01, HLA-DRB1*15:01, HLA-DRB4*01:01, HLA-DRB5*01:01 SEQIDNO:45 LKAINAQMS HLA-DRB1*01:01, HLA-DRB1*04:05, HLA-DRB1*07:01, HLA-DRB1*08:02, HLA-DRB1*09:01, HLA-DRB1*11:01, HLA-DRB4*01:01, HLA-DRB5*01:01 SEQIDNO:46 NAQMSMSHR HLA-DRB1*01:01 SEQIDNO:47 NVYVGRSLK HLA-DRB5*01:01 HLA-A*03:01 HLA-A*03 SEQIDNO:48 PKRSWNVYV HLA-DRB1*13:02, HLA-DRB1*15:01 SEQIDNO:49 QMSMSHRTM HLA-DRB1*01:01, HLA-B*15:01 HLA-B*62 HLA-DRB4*01:01 SEQIDNO:50 SLKAINAQM HLA-DRB1*04:01, HLA-B*15:01 HLA-B*62 HLA-DRB1*08:02, HLA-DRB1*09:01, HLA-DRB1*11:01, HLA-DRB1*13:02, HLA-DRB4*01:01 SEQIDNO:51 VGRSLKAIN HLA-DRB1*03:01, HLA-DRB1*04:05, HLA-DRB1*08:02, HLA-DRB1*11:01 SEQIDNO:52 VYVGRSLKA HLA-DRB1*01:01, HLA-DRB1*01:01, HLA-DRB1*03:01, HLA-DRB1*04:01, HLA-DRB1*09:01, HLA-DRB1*11:01, HLA-DRB1*13:02, HLA-DRB1*15:01, HLA-DPA1*03:01/ DPB1*04:02 SEQIDNO:53 WNVYVGRSL HLA-DRB1*01:01, HLA-DRB1*04:05, HLA-DRB1*07:01, HLA-DRB1*09:01, HLA-DRB1*15:01, HLA-DRB5*01:01 SEQIDNO:54 YVGRSLKAI HLA-DRB1*08:02, HLA-DPA1*02:01/ DPB1*01:01
TABLE-US-00003 TABLE2 LmLRAB LmLRAB Concatenation (IA,IB,IC) affinityforHLA AffinityforHLA SEQID Epitope alleleofclassI supertypesofclassII SEQIDNO:67 NENNITGGL HLA-B*18:01, HLA-B*44 HLA-B*40:01, HLA-B*40:02, HLA-B*44:02, HLA-B*44:03 SEQIDNO:68 KTKQHLREM HLA-A*30:01, HLA-A*01/HLA-A*03, HLA-A*2602 HLA-A*01 SEQIDNO:69 KARYMENNITGGLARYKTKQHVREM(longpeptide) SEQIDNO:70 RFAQGEHDI HLA-A*2403 HLA-A*24 SEQIDNO:71 RLPENAFVI HLA-A*32:01, HLA-A*01, HLA-A*02:06, HLA-A*02 HLA-A*02:11, HLA-A*02:12, HLA-A*02:16, HLA-A*02:19 SEQIDNO:72 TQGSSKAGF HLA-B*15:01, HLA-B*27, HLA-B*15:02 HLA-B*62 SEQIDNO:73 KARYRFAQGEHDIRLPENAFVIARYTQGSSKAGF(longpeptide) SEQIDNO:74 MPHVDQSSI HLA-B*07:02, HLA-B*07, HLA-B*35:01, HLA-B*08 HLA-B*51:01, HLA-B*53:01, HLA-B*08:01 SEQIDNO:75 ASFRSTEAI HLA-A*32:01, HLA-A*01, HLA-B*15:03, HLA-B*27, HLA-B*1517 HLA-B*58 SEQIDNO:76 SSIMVVANK HLA-A*30:01, HLA-A*01/HLA-A*03, HLA-A*03:01, HLA-A*01 HLA-A*11:01, HLA-A*31:01, HLA-A*68:01 SEQIDNO:77 KARYMPHVDQSSIARYASFRSTEAIRYSSIMVVANK(longpeptide) Concatenation LmLRAB-IIA affinityforHLA SEQID peptide allelesofclassII SEQIDNO:78 KGRLHSVVGRHLSIV HLA-DRB1*07:01, HLA-DRB5*01:01, HLA-DRB1*11:01, HLA-DRB1*15:01, HLA-DRB1*09:01, HLA-DRB1*01:01, HLA-DRB4*01:01 SEQIDNO:79 GRLHSVVGRHLSIVA HLA-DRB1*07:01, HLA-DRB5*01:01, HLA-DRB1*15:01, HLA-DRB1*11:01, HLA-DRB1*09:01, HLA-DRB1*01:01, HLA-DRB4*01:01 SEQIDNO:80 RLHSVVGRHLSIVAD HLA-DRB1*07:01, HLA-DRB1*15:01, HLA-DRB5*01:01, HLA-DRB1*11:01, HLA-DRB1*09:01, HLA-DRB1*01:01 SEQIDNO:81 LHSVVGRHLSIVADH HLA-DRB1*07:01, HLA-DRB1*15:01, HLA-DRB5*01:01, HLA-DRB1*11:01, HLA-DRB1*09:01, HLA-DRB1*01:01 SEQIDNO:82 HSVVGRHLSIVADHM HLA-DRB1*01:01, HLA-DRB1*15:01 SEQIDNO:83 SVVGRHLSIVADHMP HLA-DRB1*01:01, HLA-DRB1*15:01 SEQIDNO:84 VVGRHLSIVADHMPH HLA-DRB1*01:01 SEQIDNO:85 VGRHLSIVADHMPHL HLA-DRB1*03:01, HLA-DRB3*01:01, HLA-DRB1*13:02, HLA-DRB1*15:01, HLA-DRB1*01:01 SEQIDNO:86 GRHLSIVADHMPHLD HLA-DRB3*01:01, HLA-DRB1*03:01, HLA-DRB1*13:02, HLA-DRB1*01:01 SEQIDNO:87 RHLSIVADHMPLHDQ HLA-DRB3*01:01, HLA-DRB1*03:01, HLA-DRB1*13:02 Concatenation affinityforHLA AffinityforHLA Core allelesofclassII allelesofclassI SEQIDNO:88 HLSIVADHM HLA-DRB1*01:01 SEQIDNO:89 IVADHMPHL HLA-DRB1*03:01, HLA-A*02:01, HLA-DRB1*13:02, HLA-A*02:03, HLA-DRB3*01:01 HLA-A*02:06 SEQIDNO:90 LHSVVGRHL HLA-DRB1*01:01 HLA-DRB1*07:01 HLA-DRB1*09:01 HLA-DRB1*11:01 HLA-DRB1*15:01 HLA-DRB4*01:01 HLA-DRB5*01:01 SEQIDNO:91 VGRHLSIVA HLA-DRB1*01:01, HLA-DRB1*15:01 Peptide AffinityalleleclassI SEQIDNO:92 KGRLHSVVG HLA-A*30:1 SEQIDNO:93 RLHSVVGRH HLA-A*03:01,HLA-A*30:02 SEQIDNO:94 SVVGRHLSI HLA-A*02:03,HLA-A*02:06,HLA-A*32:01, HLA-B*08:01 SEQIDNO:95 VVGRHLSIV HLA-A*02:03 LongpeptideSEQID SEQIDNO:96 LHSVVGRHLSIVADHMPHLDQ SEQIDNO:97 KGRLHSVVGRHLSIVADHMPHLDQ LmlRAB-IIB ConcatenationaffinityforHLA SEQID Peptide allelesofclassII SEQIDNO:98 AIVTSRKVQEEVFDL HLA-DRB1*03:01,HLA-DRB1*11:01, HLA-DRB1*12:01 SEQIDNO:99 DLFTDVHYSEVSAKT HLA-DRB1*04:01,HLA-DRB7*01:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*01/DPB1*04:01,HLA-DPA1*02:01/ HLA-DPB1*14:01 SEQIDNO:100 EEVFDLFTDVHYSEV HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*15:01, HLA-DRB1*12:01,HLA-DRB3*01:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*01/DPB1*04:01,HLA-DPA1*02:01/ HLA-DPB1*14:01 SEQIDNO:101 ENAIVTSRKVQEEVF HLA-DRB1*03:01,HLA-DRB1*07:01, HLA-DRB1*08:02,HLA-DRB1*11:01, HLA-DRB1*12:01 SEQIDNO:102 EVFDLFTDVHYSEVS HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*12:01, HLA-DRB1*15:01,HLA-DRB3*01:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01:03/HLA-DPB1*04:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*03:01/DPB1*04:02, HLA-DPA1*01:03/HLA-DPB1*04:01, HLA-DPA1*03:01/HLA-DPB1*04:02, HLA-DPA1*02:01/HLA-DPB1*14:01 SEQIDNO:103 FDLFTDVHYSEVSAK HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*07:01,HLA-DRB1*09:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*01:03/HLA-DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*03:01/DPB1*04:02, HLA-DPA1*02:01/HLA-DPB1*14:01 SEQIDNO:104 GRSENAIVTSRKVQE HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*07:01,HLA-DRB1*11:01, HLA-DRB1*12:01,HLA-DRB5*01:01 SEQIDNO:105 IVTSRKVQEEVFDLF HLA-DRB1*03:01,HLA-DRB1*11:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*05:01, HLA-DPA1*02:01/HLA-DPB1*14:01 SEQIDNO:106 KGRSENAIVTSRKVQ HLA-DRB1*01:01,HLA-DRB1*07:01, HLA-DRB1*11:01,HLA-DRB1*12:01, HLA-DRB5*01:01 SEQIDNO:107 KVQEEVFDLFTDVHY HLA-DRB1*03:01,HLA-DRB1*04:01, HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*15:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/D?B1*01:01, HLA-DPA1*02:01/DPB1*05:01, HLA-DPA1*03:01/HLA-DPB1*04:02, HLA-DPA1*02:01/HLA-DPB1*14:01 SEQIDNO:108 LFTDVHYSEVSAKTK HLA-DRB1*01:01,HLA-DRB1*04:01, HLA-DRB1*07:01,HLA-DRB1*09:01, HLA-DRB5*01:01,HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/HLA-DPB1*14:01 SEQIDNO:109 NAIVTSRKVQEEVFD HLA-DRB1*03:01,HLA-DRB1*07:01, HLA-DRB1*08:02,HLA-DRB1*11:01, HLA-DRB1*12:01 SEQIDNO:110 QEEVFDLFTDVHYSE HLA-DRB1*03:01,HLA-DRB1*04:01, HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*15:01,HLA-DRB3*01:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*03:01/DPB1*04:02, HLA-DPA1*01:03/HLA-DPB1*04:01, HLA-DPA1*02:01/HLA-DPB1*14:01 SEQIDNO:111 RKVQEEVFDLFTDVH HLA-DRB1*03:01,HLA-DRB1*04:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*05:01, HLA-DPA1*03:01/DPB1*04:02 SEQIDNO:112 RSENAIVTSRKVQEE HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*07:01,HLA-DRB1*11:01, HLA-DRB5*01:01 SEQIDNO:113 SENAIVTSRKVQEEV HLA-DRB1*03:01,HLA-DRB1*07:01, HLA-DRB1*08:02,HLA-DRB1*11:01, HLA-DRB1*12:01 SEQIDNO:114 SRKVQEEVFDLFTDV HLA-DRB1*03:01,HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*05:01, HLA-DPA1*03:01/DPB1*04:02 SEQIDNO:115 TSRKVQEEVFDLFTD HLA-DRB1*03:01,HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*05:01, HLA-DPA1*01:03/HLA-DPB1*04:01, HLA-DPA1*03:01/HLA-DPB1*04:02, HLA-DPA1*02:01/HLA-DPB1*14:01 SEQIDNO:116 VFDLFTDVHYSEVSA HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*07:01, HLA-DRB1*15:01,HLA-DRB3*01:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*03:01/DPB1*04:02, HLA-DPA1*01:03/HLA-DPB1*04:01, HLA-DPA1*02:01/HLA-DPB1*14:01 SEQIDNO:117 VQEEVFDLFTDVHYS HLA-DRB1*03:01,HLA-DRB1*04:01, HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*15:01,HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*05:01, HLA-DPA1*03:01/DPB1*04:02 SEQIDNO:118 VTSRKQEEVFDLFT HLA-DRB1*03:01,HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*05:01, HLA-DPA1*01:03/HLA-DPB1*04:01, HLA-DPA1*03:01/HLA-DPB1*04:02, HLA-DPA1*02:01/HLA-DPB1*14:01 Concatenation affinityforHLA AffinityforHLA Core allelesofclassII allelesofclassI SEQIDNO:119 AIVTSRKVQ HLA-DRB1*11:01 SEQIDNO:120 EVFDLFTDV HLA-DPA1*01:03/ HLA-A*02:03, DPB1*02:01, HLA-A*02:06, HLA-DPA1*01/ HLA-A*68:02 DPB1*04:01, HLA-DPA1*03:01/ DPB1*04:02, HLA-DPA1*02:01/ HLA-DPB1*01:01, HLA-DPA1*01:03/ HLA-DPB1*04:01, HLA-DPA1*02:01/ HLA-DPB1*05:01 SEQIDNO:121 FTDVHYSEV HLA-DRB1*01:01, HLA-A*01:01, HLA-DRB1*07:01, HLA-A*02:01, HLA-DRB1*09:01, HLA-A*02:03, HLA-DRPA*01:03/ HLA-A*02:06, DPB1*04:01, HLA-A*35:01, HLA-DPA1*01:03/ HLA-A*68:02 DPB1*02:01 SEQIDNO:122 HYSEVSAKT HLA-DRB1*04:01 SEQIDNO:123 IVTSRKVQE HLA-DRB1*03:01, HLA-DRB1*08:02, HLA-DRB1*11:01 SEQIDNO:124 LFTDVHYSE HLA-DRB1*03:01, HLA-DRB3*01:01, HLA-DPA1*01/ DPB1*04:01, HLA-DPA1*02:01/ DPB1*01:01, HLA-DPA1*03:01/ DPB1*04:02 SEQIDNO:125 NAIVTSRKV HLA-DRB1*07:01 SEQIDNO:126 VQEEVFDLF HLA-DRB1*03:01, HLA-A*02:06 HLA-DPA1*01/ DPB1*04:01, HLA-DPA1*01:03/ DPB1*02:01, HLA-DPA1*02:01/ DPB1*01:01, HLA-DPA1*02:01/ DPB1*05:01, HLA-DPA1*03:01/ DPB1*04:02 SEQIDNO:127 YSEVSAKTK HLA-DRB1*07:01, HLA-DRB5*01:01 Concatenationaffinityfor PeptideCII HLAallelesofclassI SEQIDNO:128 DVHYSEVSA HLA-A*68*02 SEQIDNO:129 ENAIVTSRK HLA-A*68:01 SEQIDNO:130 FDLFTDVHY HLA-A*02:01,HLA-B*35:01 SEQIDNO:131 KGRSENAIV HLA-A*30:01 SEQIDNO:132 KVQEEVFDL HLA-A*02:01,HLA-A*02:06,HLA-A*32:01 SEQIDNO:133 LFTDVHYSEV HLA-A*01:01 SEQIDNO:134 SENAIVTSR HLA-A*31:01,HLA-B*44:03,HLA-A*68:01 SEQIDNO:135 TSRKDQEEV HLA-B*15:01,HLA-A*30:01,HLA-A*68:02 SEQIDNO:136 VHYSEVSAK HLA-A*03:01 Longpeptides SEQIDNO:137 SENAIVTSRKVQEEVFDLFTDVHYSEVSAKTK SEQIDNO:138 KGRSENAIVTSRKVQEEVFDLFTDVHYSEVSAKTK
TABLE-US-00004 TABLE3 PSA Concatenationaffinityfor SEQIDKC-2.6 Peptide HLAallelesofclassII SEQIDNO:139 EGYFLTDEKTSLVYG HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*11:01, HLA-DRB1*12:01,HLA-DRB1*13:02, HLA-DRB3*01:01,HLA-DRB3*02:02, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*03:01/DPB1*04:02 SEQIDNO:140 FLTDEKTSLVYGDGG HLA-DRB1*03:01,HLA-DRB3*01:01 SEQIDNO:141 GEGYFLTDEKTSLVY HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*11:01, HLA-DRB1*12:01,HLA-DRB1*13:02, HLA-DRB3*01:01,HLA-DRB3*02:02, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*03:01/DPB1*04:02, HLA-DPA1*02:01/HDPB1*14:01 SEQIDNO:142 GYFLTDEKTSLVYGD HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*07:01, HLA-DRB1*11:01,HLA-DRB3*01:01, HLA-DRB3*02:02, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*14:01, HLA-DPA1*03:01/DPB1*04:02 SEQIDNO:143 RSGEGYFLTDEKTSL HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB3*01:01, HLA-DPA1*03:01/DPB1*14:01 SEQIDNO:144 SGEGYFLTDEKTSLV HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*11:01, HLA-DRB3*01:01,HLA-DRB3*02:02, HLA-DPA1*02:01/DPB1*01:01 SEQIDNO:145 YFLTDEKTSLVYGDG HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB3*01:01, HLA-DRB3*02:02, HLA-DPA1*03:01/DPB1*04:02, HLA-DPA1*02:01/DPB1*14:01 Concatenation affinityforHLA AffinityHLA Core allelesofclassII allelesofclassI SEQIDNO:146 FLTDEKTSL HLA-DRB1*01:01, HLA-A*02:01, HLA-DRB1*03:01, HLA-A*02:03, HLA-DRB1*04:01, HLA-A*02:06, HLA-DRB1*11:01, HLA-B*39:01 HLA-DRB3*01:01, HLA-DRB3*02:02, HLA-DPA1*02:01/ DPB1*01:01, HLA-DPA1*03:01/ DPB1*04:02 SEQIDNO:147 GYFLTDEKT HLA-DRB1*04:01, HLA-DPA1*02:01/ DPB1*01:01 SEQIDNO:148 LTDEKTSLV HLA-DRB1*01:01, HLA-A*01:01, HLA-DRB1*04:01 HLA-A*02:01, HLA-A*02:03, HLA-A*02:06, HLA-A*68:02 SEQIDNO:149 RSGEGYFLT HLA-DRB1*03:01, HLA-DRB3*01:01 SEQIDNO:150 YFLTDEKTS HLA-DRB1*01:01, HLA-DRB1*04:01, HLA-DRB1*11:01 PeptideCII ConcatenationaffinityalleleCII SEQIDNO:151 ARSARSGEGY HLA-B*15:01 SEQIDNO:152 ARSGEGYFL HLA-B*39:01 SEQIDNO:153 FLTDEKTSLV HLA-A*01:01,HLA-A*02:01 SEQIDNO:154 LTDEKTSLVY HLA-A*01:01,HLA-B*15:01 SEQIDNO:155 RSARSGEGY HLA-A*30:02,HLA-B*15:01,HLA-B*57:01, HLA-B*58:01 SEQIDNO:156 RSARSGEGYF HLA-B*15:01,HLA-B*58:01 Longpeptide SEQIDNO:157 KAARSARSGEGYFLTDEKTSLVYGDGG Concatenationaffinityfor SEQIDKC-2.7 Peptide HLAallelesofclassII SEQIDNO:158 DYSHSMIRDLDFSNM HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*09:01, HLA-DRB1*12:01,HLA-DRB4*01:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*14:01 SEQIDNO:159 GSDYSHSMIRDLDFS HLA-DRB1*01:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*09:01, HLA-DRB4*01:01,HLA-DRB5*01:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*14:01 SEQIDNO:160 HSMIRDLDFSNMGLL HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*08:02, HLA-DRB1*12:01,HLA-DRB1*13:02, HLA-DRB1*15:01,HLA-DRB4*01:01, HLA-DPA1*01:03-DPB1*02:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*14:01, HLA-DPA1*03:01/DPB1*04:02 SEQIDNO:161 IRDLDFSNMGLLLSG HLA-DRB1*01:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*11:01, HLA-DRB1*12:01,HLA-DRB1*13:02, HLA-DRB1*15:01, HLA-DPA1*01:03/DPB1*04:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*14:01, HLA-DPA1*03:01/DPB1*04:02 SEQIDNO:162 MIRDLDFSNMGLLLS HLA-DRB1*01:01,HLA-DRB1*04:01, HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*12:01,HLA-DRB1*13:02, HLA-DRB1*15:01, HLA-DPA1*01:03/DPB1*04:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*14:01, HLA-DPA1*03:01/DPB1*04:02 SEQIDNO:163 MPSGSDYSHSMIRDL HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*09:01,HLA-DRB5*01:01 SEQIDNO:164 PSGSDYSHSMIRDLD HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*09:01,HLA-DRB5*01:01, LA-DPA1*02:01/DPB1*01:01 SEQIDNO:165 RDLDFSNMGLLLSGT HLA-DRB1*01:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*11:01, HLA-DRB1*12:01,HLA-DRB1*13:02, HLA-DRB1*15:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*01:03/DPB1*04:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*03:01/DPB1*04:02, HLA-DPA1*02:01/DPB1*14:01 SEQIDNO:166 SDYSHSMIRDLDFSN HLA-DRB1*01:01,HLA-DRB1*04:01, HLA-DRB1*04:05,HLA-DRB1*07:01, HLA-DRB1*12:01,HLA-DRB1*09:01, HLA-DRB4*01:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*14:01 SEQIDNO:167 SGSDYSHSMIRDLDF HLA-DRB1*01:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*09:01, HLA-DRB5*01:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*02:01/DPB1*14:01 SEQIDNO:168 SHSMIRDLDGSNMGL HLA-DRB1*03:01,HLA-DRB1*04:01, HLA-DRB1*04:05,HLA-DRB1*08:02, HLA-DRB1*12:01,HLA-DRB4*01:01 SEQIDNO:169 SMIRDLDFSNMGLLL HLA-DRB1*01:01,HLA-DRB1*03:01, HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*08:02, HLA-DRB1*12:01,HLA-DRB1*13:02, HLA-DRB1*15:01,HLA-DRB4*01:01, HLA-DPA1*01:03/DPB1*04:01, HLA-DPA1*01:03/DPB1*02:01, HLA-DPA1*02:01/DPB1*01:01, HLA-DPA1*03:01/DPB1*04:02, HLA-DPA1*02:01/DPB1*14:01 SEQIDNO:170 YSHSMIRDLDFSNMG HLA-DRB1*04:01,HLA-DRB1*04:05, HLA-DRB1*07:01,HLA-DRB1*08:02, HLA-DRB1*09:01,HLA-DRB1*12:01, HLA-DRB4*01:01 Concatenation affinityforHLA AffinityforHLA Core allelesofclassII allelesofclassI SEQIDNO:171 DFSNMGLLL HLA-DPA1*01:03/ DPB1*04:01, HLA-DPA1*03:01/ DPB1*04:02 SEQIDNO:172 FSNMGLLLS HLA-DRB1*01:01, HLA-DRB1*11:01 SEQIDNO:173 HSMIRDLDF HLA-DRB4*01:01 HLA-B*35:01, HLA-B*58:01 SEQIDNO:174 IRDLDFSNM HLA-DRB1*01:01, HLA-DRB4*01:01 SEQIDNO:175 LDFSNMGLL HLA-DRB1*07:01, HLA-DRB1*13:02, HLA-DRB1*15:01, HLA-DPA1*02:01/ DRPB*01:01, HLA-DPA1*01:03/ DPB1*02:01, HLA-DPA1*02:01/ DPB1*01:01, HLA-DPA1*03:01/ DPB1*04:02 SEQIDNO:176 MIRDLDFSN HLA-DRB1*03:01, HLA-DRB1*04:01, HLA-DRB1*04:05, HLA-DRB1*08:02 SEQIDNO:177 SDYSHSMIR HLA-DRB1*07:01, HLA-A*31:01, HLA-DRB5*01:01 HLA-A*68:01 SEQIDNO:178 YSHSMIRDL HLA-DRB1*01:01, HLA-DRB1*04:05, HLA-DRB1*07:01, HLA-DRB1*09:01, HLA-DPA1*02:01/ DPB1*01:01 ConcatenationaffinityforHLA Peptide allelesofclassI SEQIDNO:179 GSDYSHSMIR HLA-A*03:01 SEQIDNO:180 KEMPSGSDY HLA-B*15:01,HLA-A*30:02, HLA-B*44:02,HLA-B*44:03 SEQIDNO:181 MIRDLDFSNM HLA-B*15:01 SEQIDNO:182 MPSGSDYSH HLA-B*35:01,HLA-B*53:01 SEQIDNO:183 NMGLLLSGT HLA-A*02:02 SEQIDNO:184 SMIRDLDFS HLA-A*02:03,HLA-A*02:06 Longpeptide SEQIDNO:185 KEMPSGSDYSHMSIRDLDFSNMGLLLSGT
[0064] More preferentially, the sequences of these epitopes contained in the peptide compounds according to the invention are chosen from the sequences SEQ ID NO: 1 to 21, the sequences SEQ ID NO: 24 to 64, the sequences SEQ ID NO: 67, 68, 70 to 72, 74 to 76, the sequences SEQ ID NO: 78 to 95, the sequences SEQ ID NO: 98 to 136, the sequences SEQ ID NO: 139 to 156, and the sequences SEQ ID NO: 158 to 184, listed in Tables 1, 2 and 3 above, as well as the analogous, mutein and homologous derivatives thereof.
[0065] The object of the invention is a peptidic compound consisting of epitopes as described above and comprising: [0066] 1 to 21 overlapping epitopes of sequences SEQ ID
[0067] NO: 1 to 21 such as the sequences SEQ ID NO: 22 and 23, or [0068] 1 to 41 overlapping epitopes of sequences SEQ ID NO: 24 to 64 such as the sequences SEQ ID NO: 65 and 66, or [0069] (iii) 1 to 18 overlapping epitopes of sequences SEQ ID NO: 78 to 95 such as the sequences SEQ ID NO: 96 and 97, or [0070] (iv) 1 to 39 overlapping epitopes of sequences SEQ ID NO: 98 to 136 such as the sequences SEQ ID NO: 137 and 138, or [0071] (v) 1 to 18 overlapping epitopes of sequences SEQ ID NO: 139 to 156, such as the sequence SEQ ID NO: 157, or [0072] (vi) 1 to 27 overlapping epitopes of sequences SEQ ID NO: 158 to 184 such as the sequence SEQ ID NO: 185, or [0073] (viii) 1 to 3 epitopes of sequences SEQ ID NO: 67, 68, 70 to 72, 74 to 76 such as the sequences SEQ ID NO: 69, 73 and 77, optionally separated by a peptide spacer or a linking peptide sequence comprising 1 to 8 amino acids.
[0074] By way of non-limitative example of a peptidic spacer or linking sequence, mention can be made of the ARY, KGR, RY, GR, TV, K, V, Y and L units among others (Stittelaar K, et al. Vaccine, 2002, 20: 249-261; Lee Y. et al., Biomed Microdevices, 2010, 12: 207-222; Cardinaud, S., et al. Aids, 2009, 23: 1945-1954). Preferably, the peptide spacers or linking sequences used are K, KGR, RY, ARY or KARY and analogs thereof.
[0075] More preferentially, the multiepitopic peptide compounds according to the invention are chosen from the nine sequences SEQ ID NO: 23, 66, 69, 73, 77, 97, 138, 157 and 185, as well as the analogous, mutein and homologous derivatives thereof. These nine multiepitopic peptide compounds of SEQ ID NO: 23, 66, 69, 73, 77, 97, 138, 157 and 185 are for example synthesis multiepitopic peptide compounds, associated or not with peptides of the prior art having affine epitopes for the HLA molecules of class I.
[0076] The multiepitopic peptide compounds (24 to 40-mer), having a high affinity with a maximized number of HLA molecules of class I and of class II, such as the affinities for the alleles HLA of class I and HLA of class II of the epitopes in the peptides of the sequences SEQ ID NO: 22, 23, 65, 66, 69, 73, 77, 96, 97, 137, 138, and 157, 185, have a high immunoprevalent potential corresponding to an optimal vaccine coverage, in particular for the populations concerned. The degree of coverage of the peptide compounds of the invention show a very high added value compared with the peptides of the prior art. The coverage of the estimated world population with a combination of the peptide compounds according to the invention, or with the combination of the peptides B9, B10, B11, B12 and B13 (SEQ ID NO: 186 to 190), shows a very high added value of the peptides according to the invention.
[0077] The in vitro efficacy of the peptide compounds for stimulating human cells and producing cytokines of type Th1 such as IFN- has been evaluated, in combinations at 1 M for each peptide, using blood samples from individuals cured of leishmaniasis, according to a short protocol and a single stimulation. As is shown in the table in
[0078] (Pool E, G) with synthesis peptide compounds of the prior art. The values of IFN- for PHA and SLA are the values subtracted from the NS 5 days values, the values of IFN- for the Pool A to Z are the values subtracted from the values of NS 10 days and the values in bold correspond to values higher than the mean of the naive +3standard deviation. The in vitro immunogenicity of the peptide compounds is the ability of the peptides to recruit precursive lymphocytes, to induce in vitro a stimulation of specific T lymphocytes with the production of IFN-.
[0079] The immunogenicity was evaluated, with combinations of peptides (combination corresponding to the Pool H) at 1 M for each peptide, from samplings of blood of naive individuals and according to a long protocol with at least three successive stimulations. An IFN- ELISPOT test for measuring the frequency of the pre-existing naive T cells and the amplitude of the specific naive T repertoire for each peptide, was performed with the CD4+ T cells and the autologous dendritic cells of a naive individual in the presence or absence of the mixture of peptides, or in the presence of each peptide individually. Several specific lines of the peptide combination were revealed and respond specifically to at least one of the peptides of the present invention. The Pool H combination was used either with peptides not modified at the amino-terminal end and with amidation of the carboxy-terminal end, or with peptides chemically modified by a palmitoyl tail at the amino-terminal end and with amidation of the carboxy-terminal end.
[0080] The evaluation of the immunogenicity properties of peptide compounds according to the invention in naive individuals, persons who have never been in contact with the Leishmania parasite, is an important factor in predicting the efficacy of a peptide vaccine (Kwok WW et al, 2012, Frequency of epitope-specific naive CD4(+) T cells correlates with immunodominance in the human memory repertoire; Castelli F A et al, 2007, Differential capacity of T cell priming in naive donors of promiscuous CD4+ T cell epitopes of HCV NS3 and Core proteins). Immunogenicity is evaluated in vitro by measuring the frequency of the pre-existing naive T cells and the amplitude of the naive T repertoire specific for each compound. The evaluation is carried out using blood samples from naive individuals and in accordance with a long protocol (Castelli F A, et al. 2007 as aforementioned) putting purified T lymphocytes in contact with autologous dendritic cells previously incubated with the peptides of the invention. The combination of peptides used is composed of: SEQ ID NO: 23, SEQ ID NO: 66, SEQ ID NO: 73, SEQ ID NO: 77, SEQ ID NO: 97, SEQ ID NO: 138, SEQ ID NO: 157, SEQ ID NO: 185, SEQ ID NO: B11, SEQ ID NO: B12 and SEQ ID NO: B13. The compounds were synthesized chemically without amino-terminal modification and with carboxy-terminal (peptide-NH2) amidation (NH2). After three successive stimulations at intervals of seven days of the CD4+ T cells of a naive individual (MPL10; tablissement franais du sang de Toulouse), an IFN- ELISPOT was carried out with the cells in the presence or absence of the mixture of peptides, or in the presence of each compound individually. Six specific lines were revealed (p-value between 0.0269 and 0.0489 according to the specific line). Each positive line responds to at least one of the peptides of the present invention. The MPL10-3 line responds to the peptides SEQ ID NO: 73 and SEQ ID NO: 157. The MPL10-4 line responds to the peptides SEQ ID NO: 23, SEQ ID NO: 73, SEQ ID NO: 77, SEQ ID NO: 97, SEQ ID NO: 138 and SEQ ID NO: 157. The MPL10-7 line responds to the peptides SEQ ID NO: 77, SEQ ID NO: 97 and SEQ ID NO: 157. The MPL10-8 line responds to the peptides SEQ ID NO: 66, SEQ ID NO: 73, SEQ ID NO: 97, SEQ ID NO: 138 and SEQ ID NO: 157. The MPL10-10 line responds to the peptides SEQ ID NO: 66, SEQ ID NO: 73, SEQ ID NO: 77, SEQ ID NO: 97, SEQ ID NO: 138, SEQ ID NO: 157 and SEQ ID NO: 185. And the MPL10-12 line responds to the peptides SEQ ID NO: 66, SEQ ID NO: 157 and SEQ ID NO: 185.
[0081] The evaluation of the immunogenicity properties of peptide compounds of the invention in the same naive individual (MPL10) but with chemically synthesized compounds with the addition of a palmitoylated tail at the amino-terminal end and with an NH2 group at the carboxy-terminal end (PAL-peptide-NH2) was also performed. The same combination of peptides as before was used. After three stimulations with the mixture of PAL-peptide-NH2 peptides, an IFN-g ELISPOT was carried out with the cells in the presence or absence of the mixture of peptides, or in the presence of each peptide individually. Four specific lines were revealed (p-value =0.0275 for all the specific lines). The MPL10-PAL-2 line responds to the peptides SEQ ID NO: 22, SEQ ID NO: 66, SEQ ID NO: 73, SEQ ID NO: 157 and SEQ ID NO: 185. The MPL10-PAL-3 line responds to the peptides SEQ ID NO: 23, SEQ ID NO: 66, SEQ ID NO: 73, SEQ ID NO: 77, SEQ ID NO: 97, SEQ ID NO: 138, SEQ ID NO: 157 and SEQ ID NO: 185.
[0082] The invention further relates to a composition as a pharmaceutical product, for human or veterinary use, comprising at least: [0083] an epitope of a sequence chosen from among the sequences SEQ ID No: 1 to 21, the sequences SEQ ID NO: 24 to 64, the sequences SEQ ID NO: 67, 68, 70 to 72, 74 to 76, the sequences SEQ ID NO: 78 to 95, the sequences SEQ ID NO: 98 to 136, the sequences SEQ ID NO: 139 to 156, and the sequences SEQ ID NO: 158 to 184, or [0084] a peptidic compound comprising 1, 2, 3 or 4 of the epitopes of sequence SEQ ID NO: 1 to 21, SEQ ID NO: 24 to 64, SEQ ID NO: 67, 68, 70 to 72, 74 to 76, SEQ ID NO: 78 to 95, SEQ ID NO: 98 to 136, SEQ ID NO: 139 to 156, and SEQ ID NO: 158 to 184, optionally separated by a spacer as defined above, or [0085] a peptidic compound of sequence chosen from SEQ ID NO: 22, 23, 65, 66, 69, 73, 77, 96, 97, 137, 138, 157 and 185.
[0086] The invention also relates to such a composition for use thereof in prophylactic and therapeutic vaccination directed against one or more of the leishmania such as Leishmania donovani, Leishmania infantum, Leishmania chagasi, Leishmania mexicana, Leishmania amazonensis, Leishmania venezuelensis, Leishmania tropica, Leishmania major, Leishmania aethiopica, Leishmania (Viannia) braziliensis, Leishmania (Viannia) guyanensis, Leishmania(Viannia) panamensis, Leishmania (Viannia) peruviana.
[0087] Advantageously, the composition according to the invention is able to be used in prophylactic and therapeutic vaccination directed against at least three, preferably at least seven, preferably again at least ten and even more preferably against all the Leishmania listed above.
[0088] The composition advantageously comprises an adjuvant chosen from the adjuvants chosen from among the Toll-Like Receptor adjuvants of classes TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, saponins and the QA21, quilA or QS21 derivatives thereof, oil in water or water in oil emulsions, polysaccharides, cationic liposomes, virosomes or polyelectrolytes and immunomodulators chosen from sandfly saliva proteins, cytokines, peptides and heat shock proteins, for example HSP70.
[0089] Preferably, the composition according to the invention is administered subcutaneously, intradermally, by intramuscular route, intravenously, parenterally, endonasally, or by mucosal or oral route, and is therefore in a form suitable for such administrations.
[0090] An additional object of the invention relates to a composition as defined above by way of drug, vaccine or in vitro and/or in vivo diagnostic reagent, for inducing or diagnosing, in a mammal, change from a type Th2 immune state to a type Th1 immune state.
[0091] A further additional object of the invention relates to a vaccine capable of conferring cross immunoprotection vis-vis kinds of leishmaniasis. The significant antigen community shared by Leishmania makes it possible to envisage the development of a multipurpose single vaccine, consisting of highly preserved common immunogens. Targeting as a vaccine one or more antigens common to all the species of Leishmania would without any doubt represent a real advantage in terms of cross vaccination. Such a vaccine comprises at least: [0092] an epitope of sequence chosen from among the sequences SEQ ID NO: 1 to 21, the sequences SEQ ID NO: 24 to 64, the sequences SEQ ID NO: 67, 68, 70 to 72, 74 to 76, the sequences SEQ ID NO: 78 to 95, the sequences SEQ ID NO: 98 to 136, the sequences SEQ ID NO: 139 to 156, and the sequences SEQ ID NO: 158 to 184, or [0093] a peptidic compound comprising 1, 2, 3 or 4 of the epitopes of sequence SEQ ID NO: 1 to 21, SEQ ID NO: 24 to 64, SEQ ID NO: 67, 68, 70 to 72, 74 to 76, SEQ ID NO: 78 to 95, SEQ ID NO: 98 to 136, SEQ ID NO: 139 to 156, and SEQ ID NO: 158 to 184, optionally separated by a spacer as defined above, or [0094] a peptidic compound of sequence chosen from SEQ ID NO: 22, 23, 65, 66, 69, 73, 77, 96, 97, 137, 138, 157 and 185.
[0095] Such a vaccine is advantageously used for prophylactic or therapeutic vaccination directed against one or more of the Leishmania chosen from Leishmania donovani, Leishmania infantum, Leishmania chagasi, Leishmania mexicana, Leishmania amazonensis, Leishmania venezuelensis, Leishmania tropica, Leishmania major, Leishmania aethiopica, Leishmania (Viannia) braziliensis, Leishmania (Viannia) guyanensis, Leishmania (Viannia) panamensis and Leishmania (Viannia) peruviana.
[0096] The vaccine that is the object of the invention is advantageously intended for humans, canines, felines and members of the horse family. Preferentially, the vaccine that is the object of the invention is intended for humans and dogs.