WO2013186178A1 - Stimulation de la réponse immunitaire cellulaire contre le virus epstein-barr (ebv) - Google Patents

Stimulation de la réponse immunitaire cellulaire contre le virus epstein-barr (ebv) Download PDF

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WO2013186178A1
WO2013186178A1 PCT/EP2013/061929 EP2013061929W WO2013186178A1 WO 2013186178 A1 WO2013186178 A1 WO 2013186178A1 EP 2013061929 W EP2013061929 W EP 2013061929W WO 2013186178 A1 WO2013186178 A1 WO 2013186178A1
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seq
peptides
ebv
amino acid
different
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PCT/EP2013/061929
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German (de)
English (en)
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Stefan Stevanovic
Christina KYZIRAKOS
Christoph GRABENBAUER
Elina BARSAUME
Stefan AUDEHM
Hans-Georg Rammensee
Thomas Feger
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Eberhard-Karls-Universitaet Universitaetsklinikum
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Publication of WO2013186178A1 publication Critical patent/WO2013186178A1/fr

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    • 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
    • A61K39/12—Viral antigens
    • A61K39/245—Herpetoviridae, e.g. herpes simplex virus
    • 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
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A61P31/22—Antivirals for DNA viruses for herpes viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • 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
    • A61K2039/70—Multivalent vaccine
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/16011—Herpesviridae
    • C12N2710/16211—Lymphocryptovirus, e.g. human herpesvirus 4, Epstein-Barr Virus
    • C12N2710/16222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/16011—Herpesviridae
    • C12N2710/16211—Lymphocryptovirus, e.g. human herpesvirus 4, Epstein-Barr Virus
    • C12N2710/16234—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • the present invention relates to a pharmaceutical composition for stimulating the cellular immune response against the Epstein-Barr virus (EBV), a process for the preparation of such a pharmaceutical composition and peptides contained therein.
  • EBV Epstein-Barr virus
  • Epstein-Barr virus also called human herpes virus 4 (HHV 4)
  • HHV 4 human herpes virus 4
  • the main transmission path of the virus is a droplet infection or a contact or smear infection. Transmissions may also be involved in transplantations or blood transfusions.
  • the infection with the virus takes place mostly in childhood. From the age of 40, approximately 98% of people are infected with EBV. In most cases, chronic infections are asymptomatic. In 30% to 60% of all cases, however, the so-called Pfeiffer glandular fever or infectious mononucleosis may occur.
  • EBV is also causally related to lymph node cancer, Burkitt's lymphoma, Hodgkin's lymphoma and other lymphomas. Especially in immunocompromised individuals there is an increased occurrence of EBV-associated diseases.
  • EBV plays an important role in patients undergoing organ transplantation. These are immunosuppressed to prevent the rejection of the foreign organ. This promotes the multiplication of EBV and the development of the so-called lymphoproliferative disease after transplantation (English, "post-transplant lymphoproliferative disorder", PTLD).
  • PTLD post-transplant lymphoproliferative disorder
  • a targeted therapy against the Pfeiffer glandular fever does not exist so far. In most cases, there is a purely symptomatic treatment. Accordingly, the treatment methods of Hodgkin's or Burkitt's lymphoma consist of classic chemotherapy and radiation therapies. The therapy of PTLD has not been satisfactorily resolved. Often it comes here to the use of chemo and radiation therapies and various methods of immune stimulation.
  • the adoptive transfer of polyclonal T cell lines for the treatment and prevention of PTLD in hematopoietic stem cell transplantation is described, for example, in Heslop et al.
  • This object is achieved by providing a pharmaceutical composition for stimulating the cellular immune response against the Epstein-Barr virus (EBV), which has at least three different peptides each having different amino acid sequences, wherein the amino acid sequences are each selected from the group from SEQ ID No. 1 to 78.
  • EBV Epstein-Barr virus
  • the inventors have surprisingly discovered that the administration of at least three different peptides having three different amino acid sequences from the group found by the inventors leads to stimulation of the cellular immune response to EBV in almost all EBV-infected individuals.
  • the peptides provided by the inventors detect so-called "frequently recognized epitopes" (FREP) from EBV, that is epitopes of EBV, which are frequently recognized by the cellular immune system.
  • FREP frequently recognized epitopes
  • the peptides provided by the inventors capture both MHC class I and MHC class II restricted EBV specific epitopes.
  • the particular advantage of the pharmaceutical composition according to the invention is that the MHC or HLA restriction is practically overcome.
  • the pharmaceutical composition according to the invention triggers stimulation of the cellular immune response in EBV-infected individuals, regardless of which MHC or HLA profile is present. It is therefore not necessary to perform MHC or HLA typing of the affected individual prior to administration.
  • the treatment with the composition according to the invention is therefore particularly cost-effective.
  • the pharmaceutical composition may be administered directly to the individual and thus promptly induce immune stimulation.
  • the pharmaceutical composition may comprise, in addition to the peptides, a pharmaceutically acceptable carrier.
  • a pharmaceutically acceptable carrier include, for example, binders, disintegrants, fillers, lubricants and buffers, salts and other substances suitable for the formulation of medicaments; see. Rowe et al. (2006), Handbook of Pharmaceutical Excipients, 5th Edition, Pharmaceutical Press, or Bauer et al. (1999), Textbook of Pharmaceutical Technology, 6th Edition, Academictincturethane, or others.
  • the inventors have recognized that already three peptides are sufficient to capture a majority of EBV-infected individuals. This is especially true in the case of MHC class II peptides. These are not very strong in their ability to bind restrictive and can sometimes bind across alleles. The inventors were able to generate MHC class II peptides, which led to recognition rates of more than 75% among randomly selected blood donors. When using at least three MHC class II peptides according to the invention, recognition rates of approximately 100% are achieved.
  • the pharmaceutical composition requires in addition to the peptides of the invention no further active ingredients. However, it may contain other agents or potentiators.
  • adjuvants are preferred in order to non-specifically increase the immune response.
  • adjuvants include aluminum hydrochloride, MF59, AS03, monophosphoryl lipid A, etc.
  • the pharmaceutical composition has at least five different peptides according to the invention.
  • Table 1 Theoretical population coverage with increasing number of peptides at a recognition rate of the peptides of 100%; Calculation according to Schipper et al. (1996), Minimal phenotype panels. A method for achieving maximum population coverage with a minimum of HLA antigens, human immunology 51 (2), pages 95-98; Allele frequencies of the German population from www.allelefrequencies.net
  • the pharmaceutical composition has at least eight different peptides.
  • composition according to the invention with even greater safety in any EBV-infected individual triggers a cellular immune response.
  • the pharmaceutical composition according to the invention at least 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73 may have different peptides, wherein the Amino acid sequences of the various peptides are each selected from the group consisting of SEQ ID Nos. 1 to 73.
  • the pharmaceutical composition comprises at least three different MHC class II peptides each having different amino acid sequences and at least five different MHC class I peptides each having different amino acid sequences, wherein the amino acid sequences of the MHC class Il peptides are each selected from the group consisting of SEQ ID Nos. 1 to 38, and wherein the amino acid sequences of the MHC class I peptides are each selected from the group consisting of: SEQ ID Nos. 39 to 78.
  • This measure has the advantage that the MHC class I equipment and at the same time the MHC class II equipment of a human individual are detected with very high reliability, so that an optimal stimulation of the cellular immune response is ensured.
  • the pharmaceutical composition according to the invention for the treatment and / or prophylaxis of an EBV-associated disease is formed.
  • EBV-associated disease may be Pfeiffer glandular fever (infectious mononucleosis), lymph node cancer (Hodgkin's disease), Burkitt's lymphoma, or preferably the lymphoproliferative disorder after transplantation (PTLD) act.
  • PTLD lymphoproliferative disorder after transplantation
  • Another object of the present invention relates to the use of at least three, preferably at least five, more preferably at least eight different peptides, each with different amino acid sequences, for stimulating the cellular immune response against Epstein-Barr virus (EBV), wherein the amino acid sequences are each selected from the group consisting of SEQ ID Nos. 1 to 78.
  • EBV Epstein-Barr virus
  • Another object of the present invention relates to a process for the preparation of a pharmaceutical composition, the following steps (1) providing at least three, preferably at least five, more preferably at least eight different peptides, (2) formulation of the peptides into a pharmaceutically acceptable carrier, wherein the at least three, preferably at least five, more preferably at least eight different peptides each have different Amino acid sequences each selected from the group consisting of SEQ ID Nos. 1 to 78.
  • Another object of the present invention relates to a peptide for stimulating the cellular immune response against Epstein-Barr virus (EBV) or the use of a peptide for stimulating the cellular immune response against Epstein-Barr virus (EBV), wherein the peptide has an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 78.
  • Another object of the present invention relates to a peptide for stimulating the cellular immune response against the Epstein-Barr virus (EBV) or the use of a peptide for stimulating the cellular immune response to the Epstein-Barr virus (EBV), wherein the peptide has an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 4, 6, 8, 10 to 14, 16, 18, to 20, 22, 24 to 38, 48, 49, 54 , 56, 61, 62, 66, 68, 69.
  • Another object of the present invention relates to a nucleic acid molecule, which may be DNA or RNA or a genetic vector or plasmid encoding the peptide of the invention.
  • the invention also relates to a host, for example.
  • a microorganism such as a bacterium containing the nucleic acid molecule of the invention.
  • Another object of the present invention relates to a method for stimulating the cellular immune response of a living organism against EBV, which comprises the administration of the pharmaceutical composition according to the invention and / or of the peptide or peptides according to the invention in the living being.
  • a further subject of the invention is a method for the diagnostic examination of a living being for an infection with EBV, which comprises the following steps: (1) administration of the pharmaceutical composition according to the invention and / or the peptide according to the invention and / or the peptides according to the invention into the animal , (2) measuring the immune response of the livestock, (3) correlating an immune reaction of the livestock with a positive diagnosis of EBV infection.
  • the immune reaction of the living organism can be measured, for example, via the secretion of cytokine, with IFN- ⁇ being a particularly suitable marker which can be determined, for example, in the context of the so-called ELISpot assay.
  • FIG. 1 A Amplification of peptide-specific reactions in vitro leads to detectable reactions. Spot-forming units of PBMCs from eight healthy donors, measured by IFN- ⁇ ELISpot after stimulation with 10 ⁇ g ml of the peptide BLLF1 167 ex vivo and after in vitro amplification. The background (stimulation with the control peptide FlnA) was subtracted.
  • PBMCs from 16 randomly selected blood donors respond to the
  • Peptide mix 1 by IFN- ⁇ production after amplification in vitro.
  • the cells were amplified and stimulated with 10 ⁇ g ml of the peptide mix (2 g / ml per peptide) and 2 and 10 ⁇ g ml of FlnA (control peptide). Spot values of ⁇ 10, which were at least three times higher than the spot values of the control peptides, were defined as positive reactions.
  • FlnA control peptide
  • 0 medium control
  • PHA positive control phytohemagglutinin.
  • B) Stimulation with peptide mix 1 induces a multifunctional CD4 + T cell response in seropositive but not seronegative donors.
  • PBMCs from five seropositive and two seronegative donors were stimulated with 10 ⁇ g ml of peptide mix 1 after preamplification and stained intracellularly for IFN- ⁇ , TNF, granzyme B, CD154 and IL2.
  • Gating strategy live lymphocytes were selected and selected in CD4 + and CD8 + T cells. Duplets were excluded.
  • the responsive cells of the seropositive individuals are highly functional in comparison to the responsive cells of the seronegative individuals.
  • the arcs shown visualize the various functions that are simultaneously expressed by the CD4 + cells of five seropositive and two seronegative individuals. In all seropositive individuals, one-sixth to about one-third of the cells (white to mid-gray) express at least three functions. In the seronegative individuals, the reacting cells mainly show a function (black parts).
  • FIG. 4 Reactions of 16 blood donors to the peptide mix 3.
  • FIG. 5 Reactions of 4 blood donors to the peptide mixtures 4 to 13.
  • the candidate epitopes were analyzed using the database SYFPEITHI, University of Tübingen, Interfaculty Institute of Cell Biology, Germany (www.syfpeithi.de). The protein sequences were added to the database
  • PBMCs Peripheral Blood Mononuclear Cells
  • Buffy Coats or Leukapheresis products were provided by the Institute for Clinical and Experimental Transfusion Medicine, University of Tübingen, Germany.
  • the PBMCs were isolated by standard gradient separation with Ficoll (PAA) and cryopreserved in fetal calf serum (PAA) with 10% DMSO (Merck) at -80 ° C until use. Pre-amplification of specific reactions
  • PBMCs Frozen PBMCs were thawed on day 0 and incubated at high density (0.5-1 * 10 7 cells / ml) in medium (Iscove's modified Dulbecco's medium (Lonza) containing 5% heat-inactivated human serum, 50 ⁇ M ⁇ -mercaptoethanol (Roth), 1 x penicillin / streptomycin (PAA) and 25 ug ml gentamicin sulfate (Lonza)). On day 1, the peptides were added together with medium at 2-10 ⁇ g ml each. For epitope screening, the PBMCs were probed with a pool of peptides including the control peptide (Gag_H IV 296-313 or
  • IL-2 Proleukin S, Novartis
  • medium 20 U / ml The PBMCs were used on day 12 for ELISpot, intracellular cytokine stains or IFN- ⁇ secretion assays (Miltenyi Biotec). IFN-y ELISpot assays
  • Each peptide was analyzed by two- or three-fold measurements. The responses to the individual peptides were rated positive when the mean number of spots per well was at least 10 and the levels were over 3 times the mean of the negative control spots. Intracellular cytokine staining
  • the inventors determined over one hundred epitope candidates of EBV antigens from the latent and lytic phases of the virus. These were screened on T memory cells present in healthy blood donors to identify frequently recognized EBV epitopes ("frequently recognized epitopes", freps).
  • CD4 + T cells which are specific for cytomegalovirus (CMV)
  • CMV cytomegalovirus
  • the incidence of CD4 + T cells specific for epitopes of EBV is less than 1 in 10,000 PBMCs present in the ELISpot react to an epitope, usually very low.
  • preexisting specific T cells were amplified in vitro in a 12 day culture prior to reading. This resulted in an amplification of the reaction to a detectable level.
  • Each epitope candidate was screened in a first round against PBMCs from healthy blood donors by an IFN- ⁇ ELISpot. Spot values of ⁇ 10, which were at least three times higher than the spot values of the control peptides, were defined as positive reactions. Peptides that elicited positive IFN- ⁇ responses were further tested, resulting in at least 15 tested donors per peptide.
  • An exemplary ELISpot result of a tested donor with respect to a reaction against 10 different epitope candidates of the antigen BLLF1 is shown in Figure 1B.
  • Table 2 Frequently recognized EBV-specific peptides. In each case at least 16 healthy blood donors (exceptions: three each with asterisk, 12 each with two stars) with matching HLA restriction (MHC class I epitopes) or randomly selected blood donors (MHC class II epitopes) for their reactivity to the given peptide with the ELISpot assay.
  • MHC class I epitopes HLA restriction
  • MHC class II epitopes randomly selected blood donors
  • a peptide mix is recognized by over 90% of all EBV infected people.
  • Table 3 Reaction of 16 blood donors to 9 different peptides; the highlighted values correspond to positive immune reactions.
  • the epitopes EBNA3A 381 (SEQ ID NO: 13), EBNA1 514 (SEQ ID NO: 21), BMRF1 261 (SEQ ID NO: 19), BLLF1 268 (SEQ ID NO: 1) and BLLF1 167 (SEQ ID NO: 17 ) were selected because, taken together, they induced IFN- ⁇ production in all reacting donors (12/16):
  • IFN- ⁇ producing cells could be successfully stimulated in 23 of 24 randomly selected neither HLA-typed nor EBV serum status specific healthy blood donors, as detected by ELISpot. Data from 16 donors are shown in Figure 2A after amplification in vitro. The stimulated cells expressed TNF, CD154, IFNy, IL2 and granzyme B, whereas in the PBMCs of seronegative donors L13 and CG this was not detected; see. Fig. 2B.
  • the cytokine expression pattern among the responding cells was significantly more multifunctional in the seropositive donors than in the seronegative donors; see. Fig. 2C. While in the seropositive individuals tested a substantial proportion of cells (0.3 to 0.7% of CD4 + ) expressed all five activation markers, such cells could not be detected in the seronegative individuals.
  • a third mix of seven epitopes namely EBNA3A 381 (SEQ ID NO: 13), BMRF1 261 (SEQ ID NO: 19), EBNA1 514 (SEQ ID NO: 21), BLLF1 167 (SEQ ID NO: 17), BRLF1 1 19 (SEQ ID NO: 25), EBNA2 276 (SEQ ID NO: 3) and BXLF2 126 (SEQ ID NO: 7) (peptide mix 3) was recognized by 16/16 individuals tested; see. Fig. 4.
  • Peptide mix 4 BMLF1 280 (SEQ ID NO: 40), EBNA3 471 (SEQ ID NO: 47), EBNA3 247 (SEQ ID NO: 55), EBNA4 657 (SEQ ID NO: 72): Most frequent HLAs, each best peptide. Detection: 10/14, 71, 4%
  • Peptide mix 5 BRLF1 109 (SEQ ID NO: 39), BMLF1 280 (SEQ ID NO: 40), EBNA3 471 (SEQ ID NO: 47), BRLF1 148 (SEQ ID NO: 78), EBNA3 247 (SEQ ID NO: 47). 55), EBNA4 657 (SEQ ID NO: 72), EBNA6 162 (SEQ ID NO: 73): Most frequent HLAs, two best peptides each. Detection 9/14, 64.3%
  • Peptide mix 6 BRLF1 109 (SEQ ID NO: 39), EBNA3 471 (SEQ ID NO: 47), LMP2 419 (SEQ ID NO: 53), EBNA3 247 (SEQ ID NO: 55), BZLF1 190 (SEQ ID NO: 55). 57), EBN2 430 (SEQ ID NO: 66), EBNA6 162 (SEQ ID NO: 73): 7 most abundant HLAs, each best peptide. Detection 10/14, 71, 4%
  • Peptide mix 7 EBNA3 471 (SEQ ID NO: 47), EBNA4 399 (SEQ ID NO: 51), LMP2 419 (SEQ ID NO: 53), BALF2 573 (SEQ ID NO: 54), EBNA3 247 (SEQ ID NO: 53). 55), BZLF1 190 (SEQ ID NO: 57), BZLF1 173 (SEQ ID NO: 64), EBNA6 258 (SEQ ID NO: 65), EB2 430 (SEQ ID NO: 66), LMP2 200 (SEQ ID NO: 65). 71), EBNA6 162 (SEQ ID NO. 73), LMP1 256 (SEQ ID NO: 77): all but A * 02, each best peptide. Detection 13/14, 92.8%
  • Peptide mix 8 BRLF1 109 (SEQ ID NO: 39), BMLF1 280 (SEQ ID NO: 40), EBNA3 471 (SEQ ID NO: 47), BRLF1 148 (SEQ ID NO: 78), EBNA4 399 (SEQ ID NO: 47). 51), EBNA4 416 (SEQ ID NO: 52), LMP2 419 (SEQ ID NO: 53), BALF2 573 (SEQ ID NO: 54). All HLA-A, up to two best peptides. Detection 1 1/14, 78.5%.
  • Peptide mix 9 EBNA3 247 (SEQ ID NO: 55), BZLF1 190 (SEQ ID NO: 57), EBNA3 193 (SEQ ID NO: 58), EBNA4 831 (SEQ ID NO: 63), BZLF1 173 (SEQ ID NO: 58). 64), EBNA6 258 (SEQ ID NO: 65), EB2 430 (SEQ ID NO: 66), BZLF1 54 (SEQ ID NO: 67), LMP2 200 (SEQ ID NO: 71), EBNA4 657 (SEQ ID NO: 67). 72), EBNA6 162 (SEQ ID NO: 73), LMP1 156 (SEQ ID NO: 77). All HLA-B, up to two best peptides. Detection 13/14, 92.8%.
  • Peptide mix 10 BRLF1 109 (SEQ ID NO: 39), EBNA3 471 (SEQ ID NO: 47), EBNA3 247 (SEQ ID NO: 55), BZLF1 190 (SEQ ID NO: 57), BZLF1 173 (SEQ ID NO: 55). 64), EBN2 430 (SEQ ID NO: 66), EBNA1 407 (SEQ ID NO: 70). Random Mix I. Detection 13/14, 92.8%.
  • Peptide mix 1 1 BRLF1 109 (SEQ ID NO: 39), EBNA3 471 (SEQ ID NO: 47), EBNA4 399 (SEQ ID NO: 51), LMP2 419 (SEQ ID NO: 53), BALF2 573 (SEQ ID NO: 51) 54), EBNA3 247 (SEQ ID NO: 55), BZLF1 190 (SEQ ID NO: 57), BZLF1 173 (SEQ ID NO: 64), EBNA6 258 (SEQ ID NO: 65), EB2 430 (SEQ ID NO: 54) 66), LMP2 200 (SEQ ID NO: 71), EBNA6 162 (SEQ ID NO: 73), LMP1 165 (SEQ ID NO: 77). All HLAs, each best peptide. Detection 12/14, 85.7%
  • Peptide mix 12 BRLF1 109 (SEQ ID NO: 39), BMLF1 280 (SEQ ID NO: 40), LMP2 356 (SEQ ID NO: 41), LMP2 426 (SEQ ID NO: 42), LMP1 125 (SEQ ID NO: 41). 45), EBNA6 284 (SEQ ID NO: 46), EBNA3 471 (SEQ ID NO: 47), BRLF1 148 (SEQ ID NO: 78), EBNA4 399 (SEQ ID NO: 51), EBNA4 416 (SEQ ID NO: 47).
  • Peptide mix 13 BMLF1 280 (SEQ ID NO: 40), EBNA4 399 (SEQ ID NO: 51), BZLF1 (190 (SEQ ID NO: 57), EBNA4 831 (SEQ ID NO: 63), BZLF1 54 (SEQ ID NO 67), LMP2 200 (SEQ ID NO: 71). Random Mix II. Detection: 8/14, 57.1%
  • FIG. 5 shows representative ELISPOT results of donors 1755-1761 on the various mix combinations 4 to 13 after in vitro amplification. Each double row corresponds to a donor, each mix was measured as a duplicate. 1 10056: HIV-specific peptide, negative control. Row 12: one well positive control PHA and one well medium per donor.
  • the inventors provide peptides that can be used to induce stimulation of the EBV-infected individual's EBV-immune cellular immune response.
  • the peptides can be used in a peptide mixture that induces a cellular immune response to EBV in virtually any individual without prior MHC or HLA typing. This is demonstrated by the inventors using various peptide mixtures.

Abstract

Composition pharmaceutique destinée à la stimulation de la réponse immunitaire cellulaire contre le virus Epstein-Barr (EBV), comportant au moins trois peptides différents présentant chacun différentes séquence d'acides aminés, les séquence d'acides aminés étant chacune sélectionnées dans le groupe composé de SEQ ID N° 1 à 78.
PCT/EP2013/061929 2012-06-15 2013-06-10 Stimulation de la réponse immunitaire cellulaire contre le virus epstein-barr (ebv) WO2013186178A1 (fr)

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JP2022504624A (ja) * 2018-10-12 2022-01-13 ドイチェス クレブスフォルシュンクスツェントルム 拡大された抗原性スペクトルを有するエプスタイン・バーウイルス様粒子
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