US20090004214A1 - Cd4+ Survivin Epitopes and Uses Thereof - Google Patents

Cd4+ Survivin Epitopes and Uses Thereof Download PDF

Info

Publication number
US20090004214A1
US20090004214A1 US12/088,602 US8860206A US2009004214A1 US 20090004214 A1 US20090004214 A1 US 20090004214A1 US 8860206 A US8860206 A US 8860206A US 2009004214 A1 US2009004214 A1 US 2009004214A1
Authority
US
United States
Prior art keywords
peptide
hla
peptides
survivin
molecules
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US12/088,602
Other languages
English (en)
Inventor
Xiaofei Wang
Gaetan Munier
Sandra Moratille
Helene Nuyttens
Bernard Maillere
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Commissariat a lEnergie Atomique et aux Energies Alternatives CEA
Original Assignee
Commissariat a lEnergie Atomique CEA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=36570895&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=US20090004214(A1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Commissariat a lEnergie Atomique CEA filed Critical Commissariat a lEnergie Atomique CEA
Assigned to COMMISSARIAT A L'ENERGIE ATOMIQUE reassignment COMMISSARIAT A L'ENERGIE ATOMIQUE ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: NUYTTENS, HELENE, MORATILLE, SANDRA, MUNIER, GAETAN, WANG, XIAOFEI, MAILLERE, BERNARD
Publication of US20090004214A1 publication Critical patent/US20090004214A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A61K39/001148—Regulators of development
    • A61K39/00115—Apoptosis related proteins, e.g. survivin or livin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/461—Cellular immunotherapy characterised by the cell type used
    • A61K39/4611—T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/46—Cellular immunotherapy
    • A61K39/464—Cellular immunotherapy characterised by the antigen targeted or presented
    • A61K39/4643—Vertebrate antigens
    • A61K39/4644—Cancer antigens
    • A61K39/464448—Regulators of development
    • A61K39/46445—Apoptosis related proteins, e.g. survivin or livin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4747—Apoptosis related proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies

Definitions

  • the present invention relates to peptides representing CD4+ T epitopes of survivin that are able to be presented by HLA II molecules that are predominant, in particular in the Caucasian population, and to the vaccinal and diagnostic uses thereof.
  • Tumor antigens group together a collection of proteins expressed by tumor cells and that normal cells express relatively little or not at all, or which are found only in a few cell types. These antigens are divided up into five categories according to their expression profile: (1) antigens specific for the patient, resulting from point mutations related to tumorigenesis (MUM-1, CDK4, beta-catenin, HLA-A2, BCR-ABL, CASP-8), (2) tumor-specific antigens or CT (Cancer Testis) antigens, expressed in many tumors and a few normal tissues devoid of conventional HLA molecules (testicles, placenta, ovaries; MAGE, BAGE, GAGE, RAGE, NY-ESO1 antigens), (3) differentiation antigens which are expressed either during embryogenesis or in quite specific cell types (tyrosinases, gp-100, melan-A/mart-1, thyroglobulin, alpha-fetoprotein, CEA), (4) antigens overexpressed by tumors (survivin,
  • the targets which have been particularly well studied are mainly tumor-specific antigens (MAGE, NY-ESO-1).
  • MAGE tumor-specific antigens
  • these antigens may be ineffective targets for antitumor immunization, since most of them are not essential to the survival of tumor cells, which can evade immunological surveillance by decreasing or repressing the expression of these antigens.
  • Survivin (16.5 kDa), also called BIRC5 (Baculoviral IAP repeat-containing protein 5), IAP4 (inhibitor of apoptosis protein 4) or API4 (apoptosis inhibitor 4) is the smallest member of the family of apoptosis inhibitors (IAPs); it contains a zinc finger motif of 70 amino acids (BIR domain for Baculovirus inhibitor of apoptosis protein repeat), characteristic of all the members of the IAP family and a C-terminal end comprising a coiled-coil alpha-helix.
  • IAPs apoptosis inhibitors
  • the gene encoding survivin is located on human chromosome 17 (17q25) or murine chromosome 11 (11 E2); alternative splicing of a pre-mRNA comprising exons 1, 2, 2B, 3 and 4 produces three transcripts: (1) a transcript comprising exons 1,2, 3 and 4, encoding the alpha-isoform of survivin (142 amino acids, GenBank AAC51660 or SwissProt 015392), (2) a transcript comprising exons 1, 2, 2B, 3 and 4, encoding the beta-isoform or survivin-2B (165 amino acids), which results from the insertion of the sequence IGPGTVAYACNTSTLGGRGGRITR (23 amino acids, SEQ ID No. 40), between positions 74 and 75 of the sequence of the alpha-isoform, and (3) a transcript comprising exons 1, 2 and 4, encoding survivin ⁇ Ex-3.
  • Survivin is overexpressed in the majority of tumors, in particular in those associated with breast cancer, liver cancer, colon cancer, lung cancer, ovarian cancer, uterine cancer, esophageal cancer, stomach cancer, pancreatic cancer and prostate cancer, with Hodgkin's disease, with myelodisplastic syndrome with refractory anemia, and with melanomas, non-Hodgkin's lymphomas, leukemias, neuroblastomas, pheochromocytomas, soft tissue sarcomas and brain tumors.
  • the overexpression of this nonredundant protein leads to an insensitivity to apoptosis and promotes cell division.
  • This tumor antigen is a good indicator of the prognosis of cancer; a high survivin level (mRNA) is correlated with an unfavorable prognosis (Takeuchi et al., Int. J. Cancer, 2005; Rodel et al. , Cancer Res., 2005, 65,4881-4887; Muzio et al. , Cancer Lett., 2005, 225, 27-33; Kim et al., Cancer Lett., 2005, 224, 253-261).
  • CPP chromosomal passenger protein
  • INCENP inner centromere protein
  • TD60 telophase disk antigen
  • survivin represents a particularly advantageous target for antitumor immunization due to the fact that it induces a tumor-specific CD4+ and CD8+ T response, that it is expressed in most tumors and that its expression is essential to tumor survival.
  • an antigen essential to tumor survival as a target for antitumor immunization, makes it possible to avoid the problems of the tumors evading recognition by the immune system.
  • most cancers can be treated with a single vaccine, due to the fact that survivin is expressed in the majority of tumors.
  • survivin recombinant protein
  • an expression vector for this antigen or dendritic cells transfected with such an expression vector as an antitumor vaccine
  • PCT international application WO 00/03693 Pisarev et al. , Clin., Cancer Res., 2003, 9, 6523-6533; Siegel et al. , J. Immunol., 2003, 170, 5391-5397; Schaft et al., J. Immunol., 2005, 174, 3087-3097.
  • HLA-A1 major histocompatibility complex class I
  • HLA-A2-restricted peptides 95-104 and 96-1.04 (Andersen et al. , Cancer Res. , 2001, 61, 869-872; Schmitz et al., Cancer Res., 2000, 60, 4845-4849; Schmidt et al., 2003, mentioned above;
  • cytotoxic T lymphocytes have a very low frequency (of the order of 10 ⁇ 4 to 10 ⁇ 7 of the CD8+ cells; Zhang et al. , Eur. J. Immunol., 2005, 35, 776-785).
  • CTLs cytotoxic T lymphocytes
  • CD4+ T lymphocytes induce the activation of antigen-presenting cells (APCs) which, in turn, recruit tumor-specific CD8+ T lymphocytes. They are also involved in the maturation of the effector cells, namely the cytotoxic T lymphocytes.
  • APCs antigen-presenting cells
  • cytotoxic T lymphocytes the results observed in mice that do not express major histocompatibility complex class I molecules also indicate that CD4+ T lymphocytes exert a control over tumors via CTL-independent mechanisms, probably via macrophage activation.
  • CD4+ T lymphocytes can themselves be cytotoxic.
  • CD4+ T lymphocytes recognize the tumor peptides
  • HLA II molecules Human Leukocyte Antigen class II molecules.
  • the recognition may take place directly (i.e. the tumor itself presents these peptides to the T lymphocytes), but it is probable that the principal activation pathway takes place via dendritic cells.
  • These cells are in fact the main antigen-presenting cells capable of recruiting naive T lymphocytes in vivo.
  • T epitopes result from the proteolytic degradation, of the antigens by the antigen-presenting cells. They have varying lengths, generally from 13 to 25 amino acids, and have a sequence which makes them capable of binding to the HLA II molecules.
  • a peptide comprising a CD4+ T epitope is capable of stimulating, in vitro, CD4+ T cells which are specific for it, or of recruiting them in vivo. It is therefore sufficient to induce a CD4+ T response.
  • such peptides which are recognized by CD4+ T lymphocytes specific for a tumor antigen are also useful as reagents in a diagnostic test, a test for evaluating the prognosis or a test for monitoring the treatment of a cancer in humans, based on the detection of said CD4+ T cells, either directly, in particular by flow cytometry in the presence of multimers of class II molecule/peptide complexes, or indirectly, in particular by means of a lymphocyte proliferation assay, or else an antibody or cytokine assay.
  • HLA II molecules are heterodimers consisting of a polymorphic alpha-chain ( ⁇ ) and a polymorphic beta-chain ( ⁇ ).
  • ⁇ polymorphic alpha-chain
  • ⁇ polymorphic beta-chain
  • the beta-chain ( ⁇ ) encoded by the DRB1 gene which is the most widely expressed, to date has 458 alleles.
  • the two chains ( ⁇ and ⁇ ) of which they are formed are polymorphic but they have fewer alleles. 28 DQA1 alleles ( ⁇ -chain of HLA-DQ), 60 DQB1 alleles ⁇ -chain of HLA-DQ), 22 DPA1 alleles ( ⁇ -chain of HLA-DP) and 116 DPB1 alleles ( ⁇ -chain of HLA-DP) have been counted.
  • the combination between the two ⁇ - and ⁇ -chains encoded by these alleles gives rise to numerous HLA-DQ and HLA-DP molecules.
  • these isoforms have binding properties different than one another, which implies that they can bind different peptides of the same antigen.
  • the method of binding of the peptides to the HLA II molecules is more complex than that of the HLA I molecules, and no reliable programs exist for predicting the peptides capable of binding to HLA II molecules.
  • the HLA molecules have a very similar three-dimensional structure, but the binding site of HLA II molecules is a groove open at the two ends. For this reason, the HLA II molecules accept HLA II molecule-restricted peptides of variable size, generally between 13 and 25 amino acids.
  • This groove is characterized by five specificity pockets (P1, P4, P6, P7 and P9 corresponding to the positions of the residues of the peptide that they accommodate) which harbor polymorphic residues. Since the differences in sequences between the various HLA II molecules are essentially located in the pockets of the peptide-binding site, they are directly involved in the repertoire of peptides which bind to each molecule.
  • each individual recognizes in an antigen a set of peptides whose nature depends on the HLA II molecules which characterize it. Since there exists a large number of HLA II alleles, there therefore exists, in a given sequence, a large repertoire of T epitopes of very different sequences, each specific for a different allele.
  • a peptide capable of stimulating a CD4+ T response specific for a tumor antigen in some individuals may therefore be inactive in the majority of other individuals, because the latter do not recognize the tumor antigen via the same epitopes.
  • the HLA II molecules do not appear to be used uniformly for tumor recognition.
  • the T epitopes derived from tumor antigens are for the most part restricted to the HLA-DR molecules, suggesting that the HLA-DQ molecules are apparently only weakly used for tumor recognition.
  • HLA II alleles are not distributed uniformly throughout the world.
  • ten HLA-DR molecules are predominant in Europe and in the United States and cover most of the population; they are the molecules whose beta-chain is encoded by the seven alleles of the locus HLA-DRB1:*0101, *0301, *0401, *0701, *1101, *1301, *1501, and the alleles DRB3*0101, DRB4*0101 and DRB5*0101 (table I).
  • DRB1 alleles most frequent in the Caucasian population are also frequent in the African population (DRB1*0301, *0701 and *1101), the Indian population (DRB1*0301, *0701, *1301 and *1501) and the Japanese population (DRB1*1501).
  • the HLA-DRB3, -DRB4 and -DRB5 molecules which are HLA-DR molecules whose ⁇ -chain is not encoded by the DRB1 gene, are also predominant in Europe, In the USA and in Japan, since they are less polymorphic than the DRB1 molecules (table I). For example, in France, their allelic frequency is 9.2% for DRB3*0101, 28% for DRB4*0101 and 7.9% for DRB5*0101. They therefore cover by themselves 45% of the allelic frequency in France.
  • binding motifs of certain HLA-DR molecules are identical to such an extent that a peptide can bind to several HLA-DR molecules.
  • CD4+ T epitopes it is not uncommon for CD4+ T epitopes to be interwoven in one another, to such an extent that a peptide sequence may contain several CD4+ T epitopes.
  • HLA-DP4 molecules cover by themselves the majority of the allelic frequency of HLA-DP in the Caucasian population (of the order of 5 0% in Europe and 80% in North America) and are also present at not insignificant frequencies in the other populations (allelic frequency of the order of 60% in South America, 60% in India, 30% in Africa and 40% in Japan; table I).
  • Each of these DP4 molecules comprises an alpha-chain encoded either by the DPA1*0103 allele, which is the most frequent (78.2%), or by the DPA1*0201 allele (20.2%), and a beta-chain encoded by the DPB1*0401 allele (HLA-DP4 01 molecule) or the DPB1*0402 allele (HLA-DP402 molecule).
  • the HLA-DP4 molecules are entirely capable of presenting peptides to T lymphocytes insofar as numerous HLA-DP4-restricted T clones have been isolated using very varied antigens such as tetanus toxin, hepatitis B virus and the Dezmatophagoldes pteronyssimus major allergen.
  • the peptides which bind the HLA II molecules encoded by the alleles that are most frequent in the population therefore include the T epitopes of the majority of the population.
  • the inventors have identified three regions of survivin containing CD4+ T epitopes restricted to the 12 HLA II molecules predominant in the Caucasian population. These peptides represent potential candidates for prophylactic or therapeutic immunization against cancers, given that they are capable of inducing a CD4+ T response directed against the tumor, in the majority of patients immunized, since: (i) they are derived from an antigen expressed by the majority of tumor cells, (ii) they are capable of inducing CD4+ T lymphocytes specific for this antigen, and (iii) they take into account the polymorphism of the HLA II molecules.
  • these peptides which are recognized by CD4+ T lymphocytes specific for a tumor antigen expressed by the majority of tumor cells, can be used for establishing the prognosis and monitoring the evolution of cancers.
  • a subject of the present invention is a peptide derived from the alpha-isoform of survivin, for use as an antigen in the prophylactic or therapeutic immunization against cancer, or the diagnosis, prognosis or therapeutic monitoring of cancer, said peptide being selected from the group consisting of:
  • said peptides in a), b) or c), or variants in d), having a binding activity with respect to at least one HLA II molecule predominant in the Caucasian population, of less than 1000 nM, and being capable of inducing survivin-specific CD4+ T lymphocytes.
  • peptide capable of being presented by at least one HLA II molecule is intended to mean a peptide having a binding activity with respect to at least one HLA II molecule of less than 1000 nM.
  • survivin is intended to mean the three isoforms of survivin: alpha-isoform, survivin-2B and survivin ⁇ Ex-3. These isoforms may be derived from any mammal; they are preferably human isoforms.
  • the alpha-isoform of survivin corresponds to the survivin of 14 2amino acids; the positions are indicated with reference to the human sequence (Genbank AAC51660 or SwissProt O15392).
  • HLA II molecule predominant in the Caucasian population or “predominant HLA II molecule” is intended to mean an HLA II molecule comprising a beta-chain encoded by an allele whose frequency is greater than 5% in the Caucasian population, as specified in table I above.
  • cancer is intended to mean a cancer associated with the overexpression of survivin, such as, in a nonlimiting manner: breast cancer, liver cancer, colon cancer, lung cancer, ovarian cancer, uterine cancer, esophageal cancer, stomach cancer, pancreatic cancer and prostate cancer, melanomas, Hodgkin's disease, non-Hodgkin's lymphomas, leukemias, myelodysplastic syndrome with refractory anemia, neuroblastomas, pheochromocytomas, soft tissue sarcomas and brain tumors.
  • survivin such as, in a nonlimiting manner: breast cancer, liver cancer, colon cancer, lung cancer, ovarian cancer, uterine cancer, esophageal cancer, stomach cancer, pancreatic cancer and prostate cancer, melanomas, Hodgkin's disease, non-Hodgkin's lymphomas, leukemias, myelodysplastic syndrome with refractory anemia, neuroblastomas, pheochromocyto
  • the peptides according to the invention comprise a survivin CD4+ T epitope capable of being presented to at least one predominant HLA II molecule, which epitope consists of a sequence of 9 amino acids including the residues for anchoring to the HLA II molecules, flanked at one of its ends, preferably at both ends, by at least 2 amino acids, preferably 3 amino acids.
  • the peptides according to the invention have the following properties:
  • HLA II molecule-binding activity the peptides according to the invention have good affinity for the predominant HLA II molecules, in particular predominant in the Caucasian population (binding activity ⁇ 1000 nM) ; the binding activity can be measured by means of a competitive HLA II/peptide binding assay, with immunoenzymatic visualization, according to the principle described in American patent U.S. Pat. No. 6,649,166 and PCT international application WO 03/040299, for the HLA-DR and HLA-DP4 molecules, respectively;
  • these peptides are also capable of inducing survivin-specific CD4+ T cells from the precursors present in the majority of naive individuals, or else of stimulating such cells in the majority of individuals suffering from a cancer associated with the overexpression of survivin.
  • the immunogenicity of the peptides can be determined, in particular using peripheral blood mononuclear cells (PBMCs), by means of any appropriate assay known to those skilled in the art, such as, for example: a cell proliferation assay, an ELISPOT assay (assaying of cytokine-producing cells) or a test for assaying intracellular cytokines (INF- ⁇ , IL-2, IL-4 and IL-10).
  • PBMCs peripheral blood mononuclear cells
  • the invention encompasses the natural or synthetic variants obtained by mutation (insertion, deletion, substitution) of one or more amino acids in the sequence of the alpha-isoform of human survivin, provided that said variant conserves a good affinity for at least one predominant HLA II molecule, in particular predominant in the Caucasian population (binding activity ⁇ 1000 nM), and is immunogenic.
  • the natural variants result in particular from the polymorphism of survivin.
  • HLA DR molecule-binding motifs are described in particular in Sturnolio et al. , Nat. Biotech, 1999, 17, 533-534 and Rammensee et al., Immunogenetics, 1995, 41, 178-228.
  • the invention also encompasses the modified peptides derived from the previous peptides by introduction of any modification at the level of one or more amino acid residue (s), of the peptide bond or of the ends of the peptides, provided that said modified peptide conserves a good affinity for at least one predominant HLA II molecule, in particular predominant in the Caucasian population (binding activity ⁇ 1000 nM), and is immunogenic.
  • modifications which are introduced into the peptides by conventional methods known to those skilled in the art include, in a nonlimiting manner; substitution of an amino acid with a non-protein-generating amino acid (D amino acid or amino acid analogue); addition of a chemical group (lipid, oligosaccharide or polysaccharide) at a reactive function, in particular the side chain R; modification of the peptide bond (—CO—NH—), in particular with a bond of the retro or retroinverso type (—NH—CO—) or a bond other than the peptide bond; cyclization; fusion of a peptide (epitope of interest for immunization; tag that can be used for purifying the peptide, in particular in a form cleavable by a protease); fusion of the sequence of said peptide with that of a protein, in particular an ⁇ - or ⁇ -chain of an HLA II molecule or the extracellular domain of said chain, or alternatively a sequence for targeting to the endosome,
  • said HLA II molecule predominant in the Caucasian population is chosen from the HLA-DR1, HLA-DR3, HLA-DR4, HLA-DR7, HLA-DR11, HLA-DR13, HLA-DR15, HLA-DRB3, HLA-DRB4, HLA-DRB5 and HLA-DP4 molecules, preferably from the HLA-DR1, HLA-DR4, HLA-DR7, HLA-DR11, HLA-DR13, HLA-DR15, HLA-DRB5, HLA-DP401 and HLA-DP402 molecules.
  • HLA II molecules are advantageously encoded respectively by the HLA alleles DRB1*0101, DRB1*0301, DRB1*0401, DRB1*0701, DRB1*1101, DRB1*1301, DRB1*1501, DRB3*0101, DRB4*0101, DRB5*0101, DP*0401 and DP*0402.
  • HLA II molecules predominant in the Caucasian population, the HLA-DP401 and HLA-DP402 molecules are also predominant in South America, in India, in Africa and in Japan.
  • said peptide is capable of being presented by at least one HLA-DP401 or HLA-DP402 molecule.
  • said peptide is capable of being presented by at least three HLA II molecules predominant in the Caucasian population, preferably at least four HLA II molecules predominant in this population.
  • said peptide is selected from the group consisting of the peptides of 15 amino acids located between the following positions of the alpha-isoform of survivin: 17 to 31, 19 to 33, 20 to 34, 84 to 98, 90 to 104, 91 to 105, 93 to 107, 96 to 110, 99 to 113, 122 to 136 and 128 to 142; these peptides have a good affinity (binding activity ⁇ 1000 nM) for at least three HLA II molecules predominant in the Caucasian population.
  • Said peptide is advantageously selected from the group consisting of the sequences SEQ ID Nos: 5, 6, 7, 17, 19, 20, 21, 23, 24, 27 and 28.
  • said peptide is selected from the group consisting of the peptides: 17-31, 19-33, 20-34, 84-98, 90-104, 91-105, 93-107, 96-110 and 128-142 which have a good affinity (binding activity ⁇ 1000 nM) for at least four HLA II molecules predominant in the Caucasian population.
  • Such peptides make it possible both to obtain good vaccinal coverage in the individuals of the Caucasian population and to broaden the vaccinal coverage to individuals of other populations (South America, Africa, India and Japan).
  • said peptide is selected from the group consisting of the peptides: 17-31, 19-33, 90-104, 93-107, 96-110 and 128-142.
  • said peptide is selected from the group consisting of the peptides 19-33, 90-104 and 93-107.
  • said peptide has a sequence of 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 amino acids.
  • said peptide it is a variant of one of the above peptides, obtained by substitution of at least one of the residues P1, P6 and/or P9 for anchoring to the HLA II molecules, with an aromatic or hydrophobic amino acid, of the residue P6 with a cysteine (C), of the residue P9 with C, D, Q, S, T or E, and/or of the residue P4with another natural or synthetic amino acid.
  • C cysteine
  • natural or synthetic amino acid is intended to mean the 20 natural ⁇ -amino acids commonly found in proteins (A, R, N, D, C, Q, E, G, H, I, L, K, M, F, P, S, T, K, Y and V), certain amino acids rarely encountered in proteins (hydroxyproline, hydroxylysine, raethyllysine, dimethyllysine, etc.), the amino acids which do not exist in proteins, such as ⁇ -aniline, ⁇ -aminobutyric acid, homocysteine, ornithine, citrulline, canavanine, norleucine, cyclohexylalanine, etc., and also the enantiomers and the diastereoisomers of the above amino acids.
  • hydrophobic amino acid is intended to mean an amino acid selected from (one-letter code): A, V, L, I, P, W, F and M.
  • aromatic amino acid is intended to mean an amino acid selected from (one-letter code): F, W and Y.
  • said peptide it is labeled or complexed; it may in particular be complexed with labeled, for example biotinylated, HLA II molecules so as to form HLA II/peptide complexes, in particular multimeric complexes such as tetramers.
  • labeled for example biotinylated, HLA II molecules so as to form HLA II/peptide complexes, in particular multimeric complexes such as tetramers.
  • a subject of the present invention is also a polyepitopic fragment comprising the concatenation of at least two identical or different epitopes, at least one of which is a survivin CD4+ T epitope, included in a peptide as defined above, for use as an antigen in the prophylactic or therapeutic immunization against cancer or the diagnosis, prognosis or therapeutic monitoring of cancer.
  • said polyepitopic fragment has a length of 20 to 1000 amino acids, preferably of 20 to 100 amino acids.
  • Said polyepitopic fragment advantageously comprises a tag fused to one of its ends, for the purification or the detection of said fragment.
  • the tag in particular a polyhistidine sequence or a B epitope of an antigen, is preferably separated from the polyepitopic sequence by a cleavage site for a protease so as to isolate the polyepitopic sequence from the fusion.
  • said polyepitopic fragment comprises the concatenation of at least one survivin CD4+ T epitope included in the peptide as defined above and of at least one epitope selected from the group consisting of:
  • CD8+ T epitope presented by an HLA I molecule and recognized by cytotoxic T lymphocytes specific for said antigen; such CD8+ T epitopes are in particular described on the site http://www.cancerimmunity.org/peptidedatabase/tumo rspecific.htm; preferably, the CD8+ T epitope is chosen from: survivin 96-104 (LTLGEFLKL, SEQ ID No. 37) or 95-104 (LTLGEFLKL, SEQ ID No. 38), survivin-2B 80-88 (AYACNTSTL, SEQ ID No. 3 9) and the peptides as described in table I of Bachinsky et al., Cancer Immun., 2005, 5, 6;
  • a natural or synthetic, universal CD4+ T epitope such as the tetanus toxin peptide TT 830-846 (O'Sullivan et al. , J. Immunol., 1991, 147, 2663-2669), the flu virus hemaglutinin peptide HA 307-319 (O'Sullivan et al. , mentioned above), the PADRE peptide (KXVAAWTLKAA, SEQ ID No. 41; Alexander et al.
  • B epitope formed by a sugar (Alexander et al. , mentioned above), said B epitope preferably being in the form of a glycopeptide;
  • the combination of the CD4+ T epitope with at least one of the epitopes as defined above advantageously makes it possible to trigger or to modulate an antitumor immune response.
  • a subject of the present invention is also a lipopeptide comprising a peptide or a polyepitopic fragment as defined above, for use as an antigen in the prophylactic or therapeutic immunization against cancer or the diagnosis, prognosis or therapeutic monitoring of cancer.
  • Said lipopeptide is in particular obtained by addition of a lipid to an ⁇ -amino function or to a reactive function of the side chain of an amino acid of said peptide or polyepitopic fragment; it may comprise one or more chains derived from C 4 -C 20 fatty acids, optionally branched or unsaturated (palmitic acid, oleic acid, linoleic acid, linolenic acid, 2-amino-hexadecanoic acid, pimelautide, trimexautide) or a derivative of a steroid.
  • the preferred lipid portion is in particular represented by an N ⁇ -acetyllysine N ⁇ (palmitoyl) group, also called Ac-K (Pam).
  • a subject of the present invention is also a fusion protein consisting of a protein or a protein fragment, fused with a peptide or a polyepitopic fragment as defined above, for use as an antigen in the prophylactic or therapeutic immunization against cancer or the diagnosis, prognosis or therapeutic monitoring of cancer.
  • the peptide or the polyepitopic fragment may be fused with the NH2- or COOH-end of said protein, or inserted into the sequence of said protein.
  • said fusion protein consists of a peptide as defined above, fused with a sequence for targeting to the endosome, preferably derived from a human invariable chain Ii or from the LAMP-1 protein.
  • a sequence for targeting to the endosome preferably derived from a human invariable chain Ii or from the LAMP-1 protein.
  • the sequences for targeting to the endosome and their use for targeting antigens to the endosome are in particular described in Sanderson et al. (P.N.A.S., 1995, 92, 7217-7222), Wu et al. (P.N.A.S., 1995, 92, 11671-11675) and Thompson et al. (J. Virol., 1998, 72, 2246-2252).
  • said fusion protein consists of a peptide as defined above, fused with one of the chains of an HLA II molecule, preferably the beta-chain, or else with a fragment thereof corresponding to a soluble HLA II molecule, in particular a fragment corresponding to the extracellular domain preceded by the homologous signal peptide or by a heterologous signal peptide.
  • Said peptide is advantageously inserted between the signal peptide and the NH 2 -end of the extracellular domain of the ⁇ -chain, as described for the HLA-DR molecule (Kolzin et al., PNAS, 2000, 97, 291-296).
  • said peptide or said polyepitopic fragment is fused with a protein, known to those skilled in the art, for facilitating its purification or its detection, such as in particular glutathione-S-transferase (GST) and fluorescent proteins (GFP and derivatives).
  • GST glutathione-S-transferase
  • GFP fluorescent proteins
  • the sequence of the peptide or of the polyepitopic fragment of interest is preferably separated from the rest of the protein by a cleavage site for a protease, in order to facilitate the purification of said peptide or of said polyepitopic fragment.
  • a subject of the present invention is also an expression vector comprising a polynucleotide encoding a peptide, a polyepitopic fragment or a fusion protein
  • the sequence of said polynucleotide is that of the cDNA encoding said peptide or polyepitopic fragment or said fusion protein. Said sequence may advantageously be modified in such a way that the codon usage is optimal in the host in which it is expressed.
  • said polynucleotide may be linked to at least one heterologous sequence.
  • heterologous sequence relative to a nucleic acid sequence encoding survivin is intended to mean any nucleic acid sequence other than those which, naturally, are immediately adjacent to said nucleic acid sequence encoding said survivin peptide.
  • said recombinant vector comprises an expression cassette including at least one polynucleotide as defined above, under the control of appropriate regulatory sequences for transcription and, optionally, for translation (promoter, enhancer, intron, initiation codon (ATG), stop codon, polyadenylation signal).
  • nucleic acid molecule of interest in order to introduce it into and to maintain it in a eukaryotic or prokaryotic host cell
  • choice of an appropriate vector depends on the use envisioned for this vector (for example, replication of the sequence of interest, expression of this sequence, maintenance of this sequence in extrachromosomal form, or else integration into the chromosomal material of the host), and also on the nature of the host cell.
  • viral vectors such as adenoviruses, retroviruses, lentiviruses, AAVs and baculoviruses
  • said sequence isolated or inserted into a plasraid vector
  • a substance which allows it to cross the membrane of the host cells for instance a transporter such as a nanotransporter or a preparation of liposomes or of cationic polymers, or else it may be introduced into said host cell using physical methods such as electroporation or microinjection.
  • these methods may advantageously be combined, for example using electroporation combined with liposomes.
  • a subject of the present invention is also an immunogenic or vaccinal composition, characterized in that it comprises at least one peptide, one polyepitopic fragment, one fusion protein, one lipopeptide or one vector as defined above, and a pharmaceutically carrier, a carrier substance or an adjuvant.
  • the immunogenic composition according to the invention is in a pharmaceutical form suitable for parenteral (subcutaneous, intramuscular, intravenous), enteral (oral, sublingual) or local (rectal, vaginal) administration.
  • the pharmaceutically acceptable carriers, the carrier substances and the adjuvants are those conventionally used.
  • the adjuvants are advantageously chosen from the group consisting of: oily emulsions, mineral substances, bacterial extracts, saponin, alumina hydroxide, monophosphoryl-lipid A and squalene.
  • the carrier substances are advantageously selected from the group consisting of: unilamellar or multilamellar liposomes, ISCOMs, virosomes, viral pseudoparticles, saponin micelles, solid microspheres which are saccharide (poly(lactide-co-glycolide)) or gold-bearing in nature, and nanoparticles.
  • composition it comprises at least one survivin CD4+ T epitope and one survivin CD8+ T epitope in the form of a mixture of peptides, of a polyepitopic fragment and/or of an expression vector encoding said peptides or said fragment, as defined above.
  • the CD8+ T epitope is chosen from: survivin 96-104 (LTLGEFLKL) or 95-104 (LTLGEFLKL), survivin-2B 80-88 (AYACNTSTL) and the peptides as described in table I of Bachinsky et al. , Cancer Immun., 2005, 5, 6.
  • composition it comprises at least two survivin peptides including a CD4+ T epitope as defined above, selected from the group consisting of one of the following combinations:
  • peptide 17-31 and at least one of peptides 19-33, 90-104 or 128-142,
  • peptides 93-107 and 128-142 and at least one of peptides 17-31, 19-33, 96-110 or 90-104 .
  • composition it comprises a peptide including a universal CD4+ T epitope, as defined above.
  • a subject of the present invention is also the use of at least one peptide, one polyepitopic fragment, one fusion protein, one lipopeptide and/or one vector as defined above, for the preparation of a vaccine for use in the prevention or treatment of cancer.
  • the peptides according to the present invention and the derived products may be used in immunotherapy in the treatment of tumors overexpressing survivin. Said peptides or derived products are used either as a vaccine or in cell therapy, or else through a combination of the two approaches.
  • Cell therapy comprises the preparation of antigen-presenting cells (dendritic cells) by a conventional protocol comprising the isolation of peripheral blood mononuclear cells (PBMCs) from a patient to be treated and the culturing of the dendritic cells in the presence of peptide(s).
  • PBMCs peripheral blood mononuclear cells
  • the antigen-presenting cells loaded with the peptide are reinjected into the patient.
  • a subject of the present invention is also the use of at least one peptide, one polyepitopic fragment, one fusion protein, one lipopeptide and/or one vector as defined above, for the preparation of a reagent for diagnosing, evaluating the prognosis or monitoring the treatment of a cancer.
  • said reagent comprises a peptide or a fusion protein as defined above, optionally labeled or complexed, in particular complexed with labeled, for example biotinylated, HLA II molecules, in the form of multimeric complexes such as tetramers.
  • a subject of the present invention is also an in vitro method for diagnosing, evaluating the prognosis or monitoring the treatment of a cancer in an individual, characterized in that it comprises:
  • a subject of the present invention is also a kit for diagnosing, evaluating the prognosis or monitoring the treatment of a cancer, characterized in that it comprises at least one peptide as defined above and, optionally, a reagent for detecting CD4+ T lymphocytes.
  • the method according to the invention makes it possible to monitor the evolution of the CD4+ T response directed against survivin over the course of a cancer or- else of an antitumor treatment, in particular an antitumor immunotherapy;
  • the survivin-specific CD4+ T lymphocytes may be of TH1 type (secretion of IFN- ⁇ ), TH2type (secretion of IL-4) or regulatory T type (secretion of IL-10 or of TGF- ⁇ ); it is expected that the TH1-type T response is the sign of a favorable evolution of the cancer, whereas the regulatory T response is the sign of an unfavorable evolution of this cancer.
  • the detection is carried out using a biological sample containing CD4+ T cells, in particular a sample of mononuclear cells isolated from a peripheral blood sample (PBMCs).
  • PBMCs peripheral blood sample
  • the survivin-specific CD4+ T lymphocytes are detected by any means, known in themselves.
  • direct means such as flow cytometry in the presence of multimeric complexes as defined above, or else indirect means such as lymphocyte proliferation assays and assays for cytokines such as IL-2, IL-4, IL-5, IL-10 and IFN- ⁇ , in particular by immunoenzymatic techniques (ELISA, RIA, ELISPGT) or by flow cytometry (assaying of intracellular cytokines).
  • a suspension of cells (PBMCs, PBMCs depleted of CDS8+ cells, T lymphocytes pre-enriched by means of an in vitro culture step with the peptides as defined above or cloned T lymphocytes) is cultured for 3 to 5 days in the presence of said peptides and, as required, of appropriate presenting cells, such as dendritic cells, autologous or heterologous PBMCs, lymphoblastoid cells such as those obtained after infection with the EBV virus, or genetically modified cells.
  • the presence of survivin-specific CD4+ T cells in the initial suspension is detected by means of the peptides, according to one of the following methods:
  • the proliferation of the survivin-specific CD4+ T cells is measured by incorporation of tritiated thymidine into the DNA of the cells.
  • the ELISPOT assay makes it possible to reveal the presence of T cells secreting cytokines (IL-2, IL-4, IL-5, IL-10 and IFN- ⁇ ), specific for a peptide as defined above.
  • cytokines IL-2, IL-4, IL-5, IL-10 and IFN- ⁇
  • the principle of this assay is described in Czerkinsky et al., J. Immunol. Methods, 1983, 65, 109-121 and Schstoff et al. , J. Immunol. Methods, 1997, 210, 167-174, and its implementation is illustrated in international application WO 99/51630 or Gahéry-Ségard et al., J. Virol., 2000, 74, 1694-1703.
  • cytokines such as IL-2, IL-4, IL-5, IL-10 and IFN- ⁇
  • cytokines such as IL-2, IL-4, IL-5, IL-10 and IFN- ⁇
  • the principle of detection of the intracellular cytokines is described in Goulder et al., J. Exp. Med., 2000, 192, 1819-1832 and Maecker et al., J. Immunol. Methods, 2001, 255, 27-40 and its implementation is illustrated in Draenert et al., J. Immunol. Methods, 2003, 275, 19-29.
  • a biological sample preferably peripheral blood mononuclear cells (PBMCs)
  • PBMCs peripheral blood mononuclear cells
  • labeled multimeric complexes in particular labeled with a fluorochrome, formed by binding between soluble HLA II molecules and peptides as defined above, and
  • the cells labeled with said multimeric complexes are analyzed, in particular by flow cytometry.
  • the biological sample prior to the biological sample being brought into contact with said complexes, it is enriched in CD4+ T cells by bringing it into contact with anti-CD4 antibodies.
  • the HLA II/peptide multimeric complexes can be prepared from natural molecules extracted from cells expressing an HLA II molecule or from recombinant molecules produced in appropriate host cells as specified, for example, in Novak et al. (J. Clin. Investig., 1999, 104, R63-R67) or in Kuroda et al. (J. Virol., 2000, 74, 18, 8751-8756).
  • HLA II molecules may in particular be truncated (deletion of the transmembrane domain) and their sequence may be modified in order to make them soluble or else to facilitate the pairing of the alpha- and beta-chains (Novak et al. , mentioned above).
  • the loading of HLA II molecules with the peptide may be carried out by bringing a preparation of HLA II molecules as above into contact with the peptide.
  • biotinylated, soluble HLA II molecules are incubated, for 72 hours at 37° C., with a 10-fold excess as defined above, in a 10 mM phosphate-citrate buffer containing 0.15 M NaCl, at a pH of between 4.5 and 7.
  • sequence of the peptide may be introduced into one of the chains of the HLA II molecule in the form of a fusion protein which allows the preparation of HLA II/peptide multimeric complexes from appropriate host cells expressing said fusion protein. Said complexes may then be labeled, in particular with biotin.
  • the multimeric complexes of tetramer type are in particular obtained by adding, to the loaded HLA II molecules, streptavidin labeled with a fluorochrome in an amount four times less (mole for mole) with respect to the HLA II molecules, the whole mixture then being incubated for a sufficient period of time, for example overnight at ambient temperature.
  • the multimeric complexes may also be formed either by incubation of HLA II/peptide monomers with magnetic beads coupled to streptavidin, as described for HLA I molecules (Bodinier et al. , Nature, 2000, 6, 707-710), or by insertion of HLA II/peptide monomers into lipid vesicles as described for murine MHC class II molecules (Prakken, Nature Medicine, 2000, 6, 1406-1410).
  • HLA II/peptide multimeric complexes in particular of tetramer type, a suspension of cells (PBMCs, PBMCs depleted of CD8+ ceils, T lymphocytes pre-enriched by means of an in vitro culture step with peptides as defined above, or cloned T lymphocytes) is brought into contact with HLA II/peptide multimeric complexes at an appropriate concentration (for example of the order of 10 to 20 ⁇ g/ml), for a period of time sufficient to allow binding between the complexes and the survivin-specific CD4+ T lymphocytes (for example, of the order of 1 to 3 hours). After washing, the suspension is analyzed by flow cytometry: the labeling of the cells is visualized by means of the multimeric complexes which are fluorescent.
  • the flow cytometry makes it possible to separate the cells labeled with the HLA II/peptide multimeric complexes from the unlabeled cells and thus to perform cell sorting.
  • a subject of the present invention is thus also a method for sorting survivin-specific CD4+ T lymphocytes, characterized in that it comprises at least the following steps:
  • HLA II/peptide multimeric complexes in particular labeled with a fluorochrome, said complexes being formed by binding of soluble HLA II molecules with at least one peptide as defined above, and
  • a subject of the present invention is also a peptide as defined above, with the exclusion of the peptide located between positions 89 and 101 of the alpha-isoform of survivin (peptide 89-101).
  • a subject of the present invention is also a polyepitopic fragment, a fusion protein, a lipopeptide, a polynucleotide, an expression cassette, a recombinant vector and a modified prokaryotic or eukaryotic host cell, as defined above.
  • the invention encompasses in particular:
  • a) expression cassettes comprising at least one polynucleotide as defined above, under the control of appropriate regulatory sequences for transcription and, optionally, for translation (promoter, enhancer, intron, initiation codon (ATG), stop codon, polyadenylation signal), and
  • these vectors are expression vectors comprising at least one expression cassette as defined above.
  • polynucleotides, the recombinant vectors and the transformed cells as defined above can be used in particular for the production of the peptides, polyepitopic fragments and fusion proteins according to the invention.
  • the polynucleotides according to the invention are obtained by the conventional methods, known in themselves, according to standard protocols such as those described in Current Protocols in Molecular Biology (Frederick M. AUSUBEL, 2000, Wiley and son Inc, Library of Congress, USA). For example, they may be obtained by amplification of a nucleic sequence by PCR or RT-PCR, by screening genomic DNA libraries by hybridization with a homologous probe, or else by complete or partial chemical synthesis.
  • the recombinant vectors are constructed and introduced into host cells by conventional recombinant DNA and genetic engineering methods, which are known in themselves.
  • peptides and their derivatives as defined above are prepared by conventional techniques known to those skilled in the art, in particular by solid-phase or liquid-phase synthesis or by expression of a recombinant DNA in an appropriate cell system (eukaryotic or prokaryotic).
  • the peptides and their derivatives may be solid-phase synthesized according to the Fmoc technique, originally described by Merrifield et al. (J. Am. Chem. Soc., 1964, 85: 2149-), and purified by reverse-phase high performance liquid chromatography;
  • the lipopeptides may in particular be prepared according to the method described in international applications WO 99/40113 or WO 99/51630; the peptides and derivatives such as the variants, the polyepitopic fragments and the fusion proteins may also be produced from the corresponding cDNAs, obtained by any means known to those skilled in the art; the cDNA is cloned into a eukaryotic or prokaryotic expression vector and the protein or the fragment produced in the cells modified with the recombinant vector is purified by any appropriate means, in particular by affinity chromatography.
  • FIG. 1 illustrates the specificity of the CD4+ T lymphocyte lines obtained from the PBMCs of normal donors 171 and 174.
  • the CD4+ T lymphocyte lines were obtained after three stimulations one week apart, with autologous dendritic cells loaded with a mixture of the seven peptides selected (17-31,20-34, 84-98, 90-104, 93-107, 96-110 and 128-142).
  • the specificity of the T lymphocyte lines was evaluated by IFN- ⁇ BLISPOT.
  • 10 4 CD4+ T lymphocytes were incubated, in duplicate, with 10 5 autologous PBMCs, in the presence or absence of peptide. The spots were revealed 24 h after the incubation. Each bar represents the mean number of spots of the duplicates ⁇ the standard deviation. The positive values are at least three times greater than those of the negative control;
  • FIG. 2 illustrates the HLA II restriction elements of the CD4+ T lymphocyte lines specific for various survivin peptides, derived from six normal donors (169, 171, 174, 187, 188, 208).
  • the CD4+ T lymphocyte lines were obtained after three stimulations one week apart, with autologous dendritic cells loaded with a mixture of the seven peptides selected (17-31, 20-34, 84-98, 90-104, 93-107, 96-110 and 128-142).
  • the HLA II restriction of the T lymphocyte lines was evaluated by IFN- ⁇ ELISPOT.
  • FIG. 3 illustrates the presentation of survivin to the peptide-specific CD4+ T lymphocyte lines.
  • Survivin-specific CD4+ T lymphocyte lines were obtained from PBMCs from two normal donors (187 and 188).
  • Survivin (BIR5) and the HIV Nef protein were incubated for 4 hours with immature autologous dendritic cells.
  • the dendritic cells (DCs) were subsequently washed, and then, incubated with the CD4+ T lymphocytes (2 ⁇ 10 4 DCs and 10 4 CD4+ T lymphocytes).
  • the CD4+ T lymphocyte response was measured by IFN- ⁇ ELISPOT. Each bar represents the mean number of spots of the duplicates ⁇ the standard deviation;
  • FIG. 4 illustrates the response of the survivin-specific CD4+ T lymphocyte lines to decreasing doses of peptide.
  • Survivin-specific CD4+ T lymphocyte lines were obtained from PBMCs from two normal donors (174 and 188). The lines were incubated with decreasing concentrations (10 ⁇ 5 to 10 ⁇ 10 M) of peptide 17-31 ( ⁇ ), 90-104 ( ⁇ ), 93-107 ( ⁇ ), 96-110 ( ⁇ ) or 128-142 ( ⁇ ), in the presence of 10 5 autologous PBMCs.
  • the CD4+ T lymphocyte response was measured by IFN- ⁇ ELISPOT.
  • Peptides of 15 amino acids (15-mers) covering the entire survivin sequence were selected according to the presence of aromatic or hydrophobic residues at position 1 to 5, in particular at position 3 or 4, for anchoring in pocket PI of the HLA-DR and HLA-DP4 molecules.
  • sequences of the selected peptides are given in table II and the sequence listing attached in the annex.
  • the peptides were synthesized according to the Fmoc strategy by solid-phase parallel synthesis (357 MPS synthesizer, Advanced Chemtech Europe), optionally purified by RP-HPLC (Vydac C 18 column, Interchip) and controlled by mass spectrometry (ES-MS) and analytical HPLC.
  • the survivin (Bir5 or BIR5) and the HIV Nef protein are produced in E. coli, in the form of fusion proteins with glutathione-S-transferase (GST), purified on a glutathione column and separated from the GST by proteolytic cleavage.
  • GST glutathione-S-transferase
  • the assays for binding to HLA II molecules are competitive binding assays with immunoenzymatic visualization, as described in American patent U.S. Pat. No. 6,649,166, Texier et al., J. Immunol., 2000, 164,3177-3184 and Texier et al., Eur. J, Immunol., 2001, 31, 1837-1846, for the HLA-DR molecules, and in PCT international application WO 03/040299 and Castelli et al., J. Immunol., 2002, 169, 6928-6934, for the HLA-DP4 molecule.
  • the use of these assays for measuring the binding activity of peptides derived from various antigens is illustrated in American patent U.S. Pat. No. 6,649,166 and PCT international applications WO 02/090382, WO 03/040299 and WO 2004/014936.
  • HA 306-318 PYVKQNTLKLAT, SEQ ID No. 29
  • A3 152-166 EAEQLRAYLDGTGVE, SEQ ID No. 30
  • MT 2-16 AKTIAYDEEARRGLE, SEQ ID No. 31
  • B121-36 TERVRLVTRHIYNREE, SEQ ID No. 32
  • YKL AAYAAAKAAALAA
  • LOL 191-210 ESWGAVWRIDTPDKLTGPFT, SEQ ID No. 34
  • E2/E168 AGDLLAIETDKATI, SEQ ID No. 35
  • Oxy 271-287 EKKYFAATQFEPLAARL, SEQ ID No.
  • biotinylated at the NH 2 -terminal residue according to the protocol described in Texier et al., J. Immunol., 2000, 164, 3177-3184, are used as tracer under the conditions as specified in the table below.
  • results are expressed in the form of the concentration (nM) of competitive peptide which inhibits 50% of the maximum binding of the biotinylated tracer peptide (IC 50 ).
  • concentration (nM) of competitive peptide which inhibits 50% of the maximum binding of the biotinylated tracer peptide (IC 50 ).
  • the sensitivity of each assay is reflected by the IC 50 values observed with the nonbiotinylated peptides which correspond to the tracers; the IC50 values of the tracers vary by a factor of less than three.
  • a peptide having a binding activity (IC 50 ) with respect to an HLA II molecule of less than 1000 nM has a good affinity for this HLA II molecule
  • a peptide having a binding activity (IC 50 ) with respect to an HLA II molecule of less than 100 nM has a strong affinity for this HLA II molecule.
  • Three distinct regions of survivin are capable of binding with a good affinity (IC 50 ⁇ 1000 nM) to at least three predominant HLA II molecules, in particular predominant in the Caucasian population: regions 17-34 (N-terminal), 84-113 and 122-142 (C-terminal) (table IV).
  • n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. n.d. 96-110 18 5477 1666 5639 7 302 490 99-113 48 4673 2134 6462 40 9165 1200 102-116 7746 >100000 >100000 50000 8944 >100000 >100000 111-125 5000 >100000 >100000 >100000 30000 70000 122-136 5557 5809 >100000 20000 310 995 1200 128-142 8 12000 352 1342 25 >100000 284 No.
  • the peptides derived from the rest of the sequence do not exhibit any significant binding activity for at least three HLA II molecules predominant, in the Caucasian population. In addition, no peptide binds to the DR3 and DRB3 molecules.
  • Certain peptides are specific for an allele, such as the peptides 1-15, 4-18 and 52-66 which bind only to the HLA-DR11 molecule, and the peptide 41-55 which binds only to the HLA-DRB4 molecule.
  • the peptides of the N-terminal region have a good affinity for the HLA-DR1, DR7, DR11, DR13, DR15, DRB5, DP401 and DP402 molecules.
  • the binding capacities of the four overlapping peptides 84-98, 90-104, 93-107 and 96-110 cover the HLA-DR1, DR4, DR7, DR11, DR13, DR15, DRB5, DP401 and DP402 molecules.
  • Peptide 91-105 has a high affinity for the DR1, DR7, DRB5, DP401 and DP402 molecules.
  • peptide 128-142 binds with a high affinity to the DR1 and DR11 molecules and a lower affinity to the HLA-DR4 and HLA-DR15 molecules.
  • the ability of the peptides having a good affinity for the HLA II molecules predominant in the Caucasian population, to induce a stimulation of specific T lymphocytes, in vitro, was evaluated using blood samples from normal individuals (not carrying a tumor). The aim is to evaluate the ability to recruit CD4+ precursor lymphocytes, although, in a naive individual, they are at a very low frequency, i.e. to perform an in vitro immunization by means of these peptides.
  • peripheral blood mononuclear cells PBMCs
  • PBMCs peripheral blood mononuclear cells
  • SSP peripheral blood mononuclear cells
  • Olerup SSPTM HLA-DPB1 and HLA-DRB1 kit Olerup SSP AB
  • HLA-DRB1 and HLA-DPB1 alleles of these individuals are given in table V.
  • PBMCs Peripheral blood mononuclear cells
  • AIM V medium Life Technologies; 10 7 cells/ml
  • the nonadherent cells were recovered and the CD4+ T lymphocytes were then purified using anti-CD4 antibodies coupled to magnetic beads (Myltenyi Biotech kit), sorted by flow cytometry (MACS) according to the manufacturer's recommendations (Myltenyi Biotech), and frozen.
  • the adherent cells were incubated for 5 to 6 days, in AIM V medium containing 1000 u/ml of recombinant human GM-CSF and 1000 U/ml of recombinant human IL-4 (rh-GM-CSF and rh-IL-4; Tebu), and the cells that had differentiated into dendritic cells (immature dendritic cells) were subsequently cultured for 2 days in the presence of 1 ⁇ g/ml of LPS (Sigma), 1000 U/ml of rh-IL-4 and 1000 U/ml of rh-GM-CSF, so as to induce their maturation.
  • AIM V medium containing 1000 u/ml of recombinant human GM-CSF and 1000 U/ml of recombinant human IL-4 (rh-GM-CSF and rh-IL-4; Tebu)
  • rh-GM-CSF and rh-IL-4 recombinant human IL-4
  • Tebu re
  • the quality of the dendritic cell preparations is evaluated by flow cytometry (FACScalibur flow cytometerTM, Becton Dickinson) assisted by the Cell Quest ProTM software (Becton Dickinson).
  • flow cytometry FACScalibur flow cytometerTM, Becton Dickinson
  • Cell Quest ProTM software Becton Dickinson
  • the dendritic cells are labeled with anti-CD14, -CD86, -HLA-DR, -CD80 (Becton Dickinson), -Cd1a, -HLA-ABC, -CD83 and -CD16 (Beckman Coulter) antibodies, conjugated to a fluorochrome.
  • the mature dendritic cells (DCs; 5 ⁇ 10 5 cells in 1 ml) thus obtained were incubated with a mixture of peptides (10 ⁇ g of each peptide) in IMDM medium (Invitrogen) supplemented with glutamine (24 mM, Sigma), asparagines (55 mM, Sigma), arginine (150 mM, Sigma), penicillin (50 IU/ml, Invitrogen), streptomycin (50 mg/ml, Invitrogen) and 10% of human serum, hereinafter referred to as complete IMDM medium, for 4 hours at 37° C.
  • IMDM medium Invitrogen
  • glutamine 24 mM, Sigma
  • asparagines 55 mM, Sigma
  • arginine 150 mM
  • penicillin 50 IU/ml, Invitrogen
  • streptomycin 50 mg/ml, Invitrogen
  • 10% of human serum hereinafter referred to as complete IMDM medium
  • the mature dendritic cells were subsequently washed and then incubated, in the presence of previously thawed autologous CD4+ T lymphocytes (10 4 DCs and 10 5 CD4+ T), in 200 ⁇ l of complete IMDM medium containing 1000 U/ml of IL-6 (R&D systems) and 10 ng/ml of IL-12 (R&D systems), After 7 days (D7), the culture was stimulated a first time, by means of mature dendritic cells previously thawed and loaded with the mixture of peptides, in the medium containing IL-2 (10 U/ml) and IL-7 (5 ng/ml).
  • IL-2 10 U/ml
  • IL-7 5 ng/ml
  • HLA-DP4-restricted survivin-specific CD4+ T lymphocyte lines were produced using the mixture of peptides 84-98, 93-107, 90-104 and 19-33 for the stimulation of the autologous dendritic cells, according to a protocol similar to that use for the production of T lymphocyte lines restricted to the set of HLA-DR molecules predominant in the Caucasian population.
  • the anti-IFN- ⁇ human monoclonal antibody 1-D1K (MABTECH), diluted to 10 ⁇ g/ml in PBS buffer, was adsorbed onto nitrocellulose plates (Multiscreen HA;
  • the antigen-presenting cells are autologous PBMCs, immature autologous dendritic cells prepared as specified above, or a mouse fibroblast line (line L), transfected with the cDNA encoding one of the HLA-DR or HLA-DP4 molecules to be tested (Yu et al. Hum. Immunol. 1990, 27, 132-135), so as to verify the specificity of the lines with respect to the HLA-DR and HLA-DP4 molecules.
  • the antigen-presenting cells (10 5 autologous PBMCs, 3 ⁇ 10 4 transfected L cells or 2 ⁇ 10 4 immature autologous dendritic cells) and the CD4+ T lymphocytes to be tested (10 4 CD4+ T lymphocytes) were subsequently added to the plates and incubated for 24 h at 37° C., in the presence or absence of a single peptide (2 ⁇ g), of a mixture of peptides (2 ⁇ g of each peptide) or of a protein (Bir5 or Nef).
  • the proteins (1 ⁇ M) are incubated for 4 h at 37° C. in the presence of immature dendritic cells, and then washed.
  • the peptides are added directly to the plates.
  • the peptides are used at various concentrations ranging from 10 ⁇ 5 to 10 ⁇ 10 M. After three successive washes with water, and then with PBS buffer-0.05% Tween and, finally, with PBS alone, 100 ⁇ l of biotin-conjugated anti-IFN- ⁇ secondary antibody (7-B6-1-biotin, MABTECH), diluted to 0.25 ⁇ g/ml in PBS containing 1% BSA, were added to each well. After incubation for one hour, the plates were washed again and incubated with 100 ⁇ l/well of extravidin-phosphatase (E-2636, SIGMA), diluted to 1/6000.
  • E-2636 extravidin-phosphatase
  • survivin-specific CD4+ T lymphocyte lines were obtained from seven normal donors (table VI). The specificity of the lines for the survivin peptides was analyzed by IFN- ⁇ ELISPOT using autologous PBMCs as antigen-presenting cells.
  • T lymphocyte lines No. of T lymphocyte lines peptide Individual 17-31 20-34 84-98 90-104 93-107 96-110 128-142 D169 0 0 0 1 1 12 0 D171 2 1 3 3 0 0 7 D174 10 0 0 3 2 1 1 D179 1 0 2 2 0 2 4 D187 4 6 0 1 0 2 1 D188 1 2 3 7 3 1 0 D180 0 0 3 2 2 1 1 Total 18 9 11 19 8 19 14 Frequency 5/7 3/7 4/7 7/7 4/7 6/7 5/7 of responders
  • FIG. 1 shows the results obtained with 12 CD4+ T lymphocyte lines derived from normal donors 171 and 174. These lines are stimulated with the peptide mixture presented by autologous dendritic cells but in the absence of the mixture. Among the 60 wells seeded for each donor, 12 and 13 peptide-specific T lymphocyte lines were obtained, respectively for donor 171 and donor 174, suggesting that these lines derive from a few T lymphocyte precursors which were amplified by the weekly stimulations with the peptides. In fact, the T lymphocyte lines are generally specific for a single peptide or for two overlapping peptides, suggesting that they are specific for the region common to the two peptides ( FIG. 1 ). All the peptides induced at least one line (table VI and FIG. 1 ).
  • the seven donors cover the HLA-DR haplotypes predominant in the population (table VI), namely: HLA-DR1, -DR4, -DR7, -DR1, -DR13 and -DR15 and the corresponding second DR molecule (DRB3, DRB4 and DRB5).
  • HLA-DR molecules predominant in the Caucasian population, only the HLA-DR3 molecule is absent.
  • Five donors have an HLA-DP4 molecule, in accordance with the frequency of this molecule in the Caucasian population.
  • the peptide mixture induces specific CD4+ T lymphocyte lines in each donor, although the donor sampling was selected so as to include multiple HLA II haplotypes.
  • Peptide 17-31 induces 18 T lymphocyte lines
  • peptide 20-34 induces 9 lines
  • peptide 84-98 induces 11 lines
  • peptide 90-104 induces 19 lines
  • peptide 93-107 induces 8 lines
  • peptide 96-110 induces 19 lines
  • peptide 128-142 induces 14 lines.
  • Peptide 90-104 is immunogenic in all the donors and peptides 17-31, 96-110 and 128-142 induced T lymphocyte lines in five or six of the seven donors.
  • the restriction elements of the HLA II molecules responsible for the specific stimulation of the CD4+ T lymphocytes by the peptides was evaluated for 19 different CD4+ T lymphocyte lines, by IFN- ⁇ 0 ELISPOT, in the presence of L cells trarsfected with one of the HLA-DR molecules or with the HLA-DP4 molecule, or in the absence of presenting cells.
  • the lines are induced specifically by the presentation of the peptide by the antigen-presenting cells, given that no IFN- ⁇ production is detected in the absence of peptide ( FIG. 2 ) and that a modest stimulation is observed in the absence of presenting cells.
  • FIG. 2 illustrates the results obtained for 12 of the lines tested.
  • Most of the T lymphocyte lines are stimulated by their peptide presented by a single HLA II molecule. However, a degeneration of the recognition is observed, for example for the 174.31 and 187.34 lines. The stimulation does not result from the presentation of the peptides by activated neighboring T lymphocytes, since only a modest stimulation is observed in the absence of L cells.
  • DR7, DR15, DP4 and DRB4 are restriction elements for peptides 17-31; DP4 is a restriction element for peptide 20-34, DR7, DR11, DRB5 and DP4 are restriction elements for peptide 90-104, DR11 and DP4 are restriction elements for peptide 93-107, DR4, DR7 and DRB4 are restriction elements for peptide 96-110, and DR15 is a restriction element for peptide 128-142.
  • the ability of seven peptide-specific T lymphocyte lines derived from two different donors, to recognize the native protein was also analyzed. The results given in FIG. 3 are representative of these experiments.
  • the two T lymphocyte lines (187.54 and 187.34) specific for peptides 17-31 and 20-34 specifically recognize the autologous dendritic cells, preloaded with recombinant survivin. They are not stimulated in the presence of dendritic cells that are not loaded or dendritic cells that are loaded with the HIV Nef protein.
  • the T lymphocyte lines 188.19 and 188.57 are stimulated by peptides 90-104 and 93-107, alone or as a mixture, and specifically recognize the survivin protein.
  • HLA-DP4-restricted lines specific for a survivin peptide were demonstrated; these lines are stimulated by the peptide mixture presented by HLA-DP4, but in the absence of the mixture (negative control) or of HLA-DP4 (control without APCs, table VII).
  • Each line responds to one of the peptides of the mixture.
  • three are immunogenic in an HLA-DP4 context (peptides 93-107, 90-104 and 19-33).

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Immunology (AREA)
  • Organic Chemistry (AREA)
  • Epidemiology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Oncology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Cell Biology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
US12/088,602 2005-09-30 2006-09-28 Cd4+ Survivin Epitopes and Uses Thereof Abandoned US20090004214A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
FR0510013 2005-09-30
FR0510013A FR2891462B1 (fr) 2005-09-30 2005-09-30 Epitopes t cd4+ de la survivine et leurs applications
PCT/FR2006/002196 WO2007036638A1 (fr) 2005-09-30 2006-09-28 Epitopes t cd4+ de la survivine et leurs applications

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/FR2006/002196 A-371-Of-International WO2007036638A1 (fr) 2005-09-30 2006-09-28 Epitopes t cd4+ de la survivine et leurs applications

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US14/510,550 Continuation US9687539B2 (en) 2005-09-30 2014-10-09 CD4+ T survivin epitopes and uses thereof

Publications (1)

Publication Number Publication Date
US20090004214A1 true US20090004214A1 (en) 2009-01-01

Family

ID=36570895

Family Applications (2)

Application Number Title Priority Date Filing Date
US12/088,602 Abandoned US20090004214A1 (en) 2005-09-30 2006-09-28 Cd4+ Survivin Epitopes and Uses Thereof
US14/510,550 Active US9687539B2 (en) 2005-09-30 2014-10-09 CD4+ T survivin epitopes and uses thereof

Family Applications After (1)

Application Number Title Priority Date Filing Date
US14/510,550 Active US9687539B2 (en) 2005-09-30 2014-10-09 CD4+ T survivin epitopes and uses thereof

Country Status (8)

Country Link
US (2) US20090004214A1 (fr)
EP (1) EP1931377B1 (fr)
JP (1) JP5452923B2 (fr)
CA (1) CA2624498C (fr)
DK (1) DK1931377T3 (fr)
ES (1) ES2510493T3 (fr)
FR (1) FR2891462B1 (fr)
WO (1) WO2007036638A1 (fr)

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100029571A1 (en) * 2008-05-14 2010-02-04 Immatics Biotechnologies Gmbh Novel and powerful mhc-class ii peptides derived from survivin
US20110027303A1 (en) * 2008-03-31 2011-02-03 Bioimmulance Co., Ltd. Partial peptide of survivin presented on mhc class ii molecule and use therof
US20160218021A1 (en) * 2015-01-27 2016-07-28 Advanced Semiconductor Engineering, Inc. Semiconductor package and method of manufacturing the same
CN109600991A (zh) * 2016-03-21 2019-04-09 万可思乐研究股份有限公司 包含存活素肽的免疫原性组合物
US10738278B2 (en) 2014-07-15 2020-08-11 Juno Therapeutics, Inc. Engineered cells for adoptive cell therapy
US10786533B2 (en) 2015-07-15 2020-09-29 Juno Therapeutics, Inc. Engineered cells for adoptive cell therapy
CN113329761A (zh) * 2018-09-04 2021-08-31 特雷斯生物有限公司 肽疫苗

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2830940B1 (fr) 2001-10-17 2007-06-15 Commissariat Energie Atomique Procede de selection de ligands d'hla-dp4 et ses applications
US20140341939A1 (en) * 2011-12-14 2014-11-20 National University Corporation Kochi University Modification of helper t cell-inducing polypeptide
FR3008099B1 (fr) 2013-07-05 2020-08-07 Commissariat Energie Atomique Peptides immunogenes de l'antigene tumoral cycline b1
EP3129034A4 (fr) * 2014-03-14 2017-11-22 Brian J. Czerniecki Méthodes de suivi de la réponse lymphocytaire impliquant des lymphocytes t auxiliaires cd4+ de type 1 dans le cancer et la reconstitution immunitaire
WO2016153761A1 (fr) * 2015-03-13 2016-09-29 Czerniecki Brian J Méthodes de suivi de la réponse lymphocytaire impliquant des lymphocytes t auxiliaires cd4+ de type 1 dans le cancer et la reconstitution immunitaire
US10414810B2 (en) 2015-05-07 2019-09-17 H. Lee Moffitt Cancer Center And Research Institute, Inc. Double mutant survivin vaccine
MA47678A (fr) 2017-03-03 2021-05-26 Treos Bio Ltd Plateforme personnalisée d'identification de peptide immunogène
FR3087448B1 (fr) 2018-10-23 2023-10-13 Pdc Line Pharma Lignee pdc modifiee pour secreter une cytokine
FR3090319A1 (fr) 2018-12-21 2020-06-26 Commissariat A L'energie Atomique Et Aux Energies Alternatives Melanges d’epitopes t cd8+ immunogenes de la cycline b1

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5840839A (en) * 1996-02-09 1998-11-24 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Alternative open reading frame DNA of a normal gene and a novel human cancer antigen encoded therein

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000003693A1 (fr) * 1998-07-14 2000-01-27 Jenner Biotherapies, Inc. Survivine et certains de ses peptides envisages comme vaccin anticancereux
FR2830940B1 (fr) * 2001-10-17 2007-06-15 Commissariat Energie Atomique Procede de selection de ligands d'hla-dp4 et ses applications
US7892559B2 (en) * 2002-01-30 2011-02-22 Survac Aps Survivin-derived peptides and use thereof
WO2004055183A2 (fr) * 2002-12-17 2004-07-01 Fondazione Centro San Raffaele Del Monte Tabor Epitope immunodominant specifique d'un antigene carcino-embryonnaire reconnu par les lymphocytes t cd4+ et utilisations dudit epitope
US20070104689A1 (en) * 2005-09-27 2007-05-10 Merck Patent Gmbh Compositions and methods for treating tumors presenting survivin antigens

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5840839A (en) * 1996-02-09 1998-11-24 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Alternative open reading frame DNA of a normal gene and a novel human cancer antigen encoded therein

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10172925B2 (en) 2008-03-31 2019-01-08 tella, Inc. Uses of partial peptides of survivin and variations thereof
US20110027303A1 (en) * 2008-03-31 2011-02-03 Bioimmulance Co., Ltd. Partial peptide of survivin presented on mhc class ii molecule and use therof
AU2009232774B2 (en) * 2008-03-31 2014-05-29 Tella Inc. Partial peptide of Survivin presented on MHC class II molecule and use thereof
US8647629B2 (en) 2008-05-14 2014-02-11 Immatics Biotechnologies Gmbh Powerful MHC-class II peptides derived from survivin
US9498512B2 (en) 2008-05-14 2016-11-22 Immatics Biotechnologies Gmbh Powerful MHC-class II peptides derived from survivin
US20100029571A1 (en) * 2008-05-14 2010-02-04 Immatics Biotechnologies Gmbh Novel and powerful mhc-class ii peptides derived from survivin
US10434136B2 (en) 2008-05-14 2019-10-08 Immatics Biotechnologies Gmbh Powerful MHC-class II peptides derived from survivin
US11957729B2 (en) 2008-05-14 2024-04-16 Immatics Biotechnologies Gmbh Powerful MHC-class II peptides derived from survivin
US10738278B2 (en) 2014-07-15 2020-08-11 Juno Therapeutics, Inc. Engineered cells for adoptive cell therapy
US20160218021A1 (en) * 2015-01-27 2016-07-28 Advanced Semiconductor Engineering, Inc. Semiconductor package and method of manufacturing the same
US10786533B2 (en) 2015-07-15 2020-09-29 Juno Therapeutics, Inc. Engineered cells for adoptive cell therapy
CN109600991A (zh) * 2016-03-21 2019-04-09 万可思乐研究股份有限公司 包含存活素肽的免疫原性组合物
CN113329761A (zh) * 2018-09-04 2021-08-31 特雷斯生物有限公司 肽疫苗

Also Published As

Publication number Publication date
FR2891462B1 (fr) 2009-10-16
EP1931377B1 (fr) 2014-07-02
US20150050304A1 (en) 2015-02-19
EP1931377A1 (fr) 2008-06-18
FR2891462A1 (fr) 2007-04-06
ES2510493T3 (es) 2014-10-21
JP2009509518A (ja) 2009-03-12
WO2007036638A1 (fr) 2007-04-05
JP5452923B2 (ja) 2014-03-26
CA2624498C (fr) 2016-07-05
DK1931377T3 (da) 2014-10-13
US9687539B2 (en) 2017-06-27
CA2624498A1 (fr) 2007-04-05

Similar Documents

Publication Publication Date Title
US9687539B2 (en) CD4+ T survivin epitopes and uses thereof
CA2546794C (fr) Proteines appartenant a la famille bcl-2, fragments associes, et utilisation therapeutique de ces proteines et fragments sur des patients atteints du cancer
CA2881594C (fr) Composition de vaccin contre le cancer
US20110159022A1 (en) Immunogenic Peptides Derived from the Midkine Protein, as an Anticancer Vaccine
EP2234635A2 (fr) Immunothérapie basée sur la rhoc
JP2003512057A (ja) Ｍａｇｅ−ａ１２抗原ペプチド及びその利用
WO2007026078A2 (fr) Epitopes t cd4+ des antigenes mage-a restreints a hla-dp4 et leurs applications
US10550166B2 (en) Immunogenic peptides of the cyclin B1 tumor antigen
EP3532487A1 (fr) Nouveaux composés pdl2

Legal Events

Date Code Title Description
AS Assignment

Owner name: COMMISSARIAT A L'ENERGIE ATOMIQUE, FRANCE

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:WANG, XIAOFEI;MUNIER, GAETAN;MORATILLE, SANDRA;AND OTHERS;REEL/FRAME:021343/0389;SIGNING DATES FROM 20080529 TO 20080610

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION