Epstein-barr-virus vaccine

Abstract

The present invention relates to a vaccine comprising a particle, said particle comprising (i) at least one Epstein-Barr virus (EBV) structural polypeptide, (ii) at least one EBV lytic polypeptide, (iii) membrane lipids, said particle being devoid of EBV DNA, wherein (a) the B-cell transformation capacity of one or more EBV polypeptides required for B-cell transformation as comprised in said particle is disabled while their immunogenicity is maintained; and/or (b) said particle is devoid of one or more EBV polypeptides required for B-cell transformation. Furthermore, the invention relates to a method for generating a particle, to a cell obtained in the method of the invention, a kit comprising the vaccine or the particle generated according to the method of the invention. Also, the invention relates to the use of the vaccine or the particle generated according to the method of the invention for generating CD8+ cells specific for an EBV antigen.

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         5 . The vaccine of any one of  claims 1 ,  2  or  4  or the method of  claim 3  or  4 , wherein the one or more EBV polypeptides required for B-cell transformation which are lacking according to (b) of  claim 1  or (ab) of  claim 3  are selected from the group consisting of EBNA-2, EBNA-3a, EBNA-3b and EBNA-3c. 
     
     
         6 . (canceled) 
     
     
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         12 . (canceled) 
     
     
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         15 . A vaccine comprising,
 (a) at least one Epstein-Barr virus (EBV) polypeptide selected from the group consisting of EBV structural polypeptides, EBV lytic polypeptides and EBV latent polypeptides, and   (b) at least one membrane lipid, wherein the vaccine lacks any detectable EBV DNA.   
     
     
         16 . The vaccine of  claim 15 , comprising at least two EBV polypeptides. 
     
     
         17 . The vaccine of  claim 16 , wherein said at least two EBV polypeptides are selected independently from the group consisting of EBV structural polypeptides and EBV lytic polypeptides. 
     
     
         18 . The vaccine of  claim 17  wherein the B-cell transformation capacity of any of said at least two EBV polypeptides is disabled while their immunogenicity is maintained. 
     
     
         19 . The vaccine of  claim 18 , which is devoid of EBV polypeptides required for B-cell transformation. 
     
     
         20 . The vaccine of  claim 18 , wherein said EBV structural polypeptides are selected from the group consisting of membrane polypeptides, tegument polypeptides, capsid polypeptides, and combinations thereof. 
     
     
         21 . The vaccine of  claim 20 , wherein said membrane polypeptides are selected from the group consisting of BALF4, BLLF1 (also termed gp350), BDLF2, BDLF3, BKRF2, BLRF1, BNLF1 (also termed LMP-1), TP (also termed LMP-2a), BXLF2, BZLF2 and any combination thereof. 
     
     
         22 . The vaccine of  claim 21 , wherein said lytic polypeptides are selected from the group comprising the immediate early genes, the early genes and the late lytic genes BZLF1, BRLF1, BMRF1, BMLF1, BALF2, BALF5, BGL2, BHRF1, BALF4, BDLF3 and any combination thereof. 
     
     
         23 . The vaccine of  claim 22 , wherein said EBV structural polypeptide is BLLF1 (also termed gp350) and said EBV lytic polypeptide is BZLF1. 
     
     
         24 . The vaccine of  claim 16 , comprising at least three EBV polypeptides, wherein at least one of said at least three EBV polypeptides is selected from each of the groups consisting of EBV structural polypeptides, EBV lytic polypeptides and EBV latent polypeptides. 
     
     
         25 . The vaccine of  claim 18 , wherein one of the at least two EBV polypeptides whose transformation capacity is disabled is LMP-1. 
     
     
         26 . The vaccine of  claim 19 , wherein said vaccine lacks any member selected from the group consisting of EBNA-2, EBNA-3a, EBNA-3b and EBNA-3c. 
     
     
         27 . The vaccine of  claim 15  further comprising at least one member selected from the group consisting of non-EBV viral polypeptides, non-viral polypeptides, non-EBV viral nucleic acid sequences, non-viral nucleic acid sequences and vaccine adjuvants. 
     
     
         28 . A method for generating a vaccine particle, said method comprising the steps of:
 (a) transfecting a cell with a modified EBV genome, wherein said modified EBV genome in comparison to a wildtype EBV genome:
 i. lacks one or more sequences that are required for the packaging of said wildtype EBV genome, lacks one or more sequences encoding EBV polypeptides required for said packaging, or comprises one or more sequences encoding EBV polypeptides whose packaging capacity is disabled, 
 ii. lacks one or more sequences encoding EBV polypeptides that are required for B-cell transformation or comprises one or more sequences encoding EBV polypeptides whose B-cell transformation capacity is disabled; and 
 iii. lacks one or more sequences encoding EBV polypeptides that are required for inducing replication of an EBV and/or comprises one or more sequences encoding EBV polypeptides whose capacity for inducing EBV replication is disabled; 
   (b) culturing the cell obtained in step (a) under conditions that allow expression of said modified EBV genome;   (c) inducing the replicative phase of EBV; and   (d) isolating said vaccine particle.   
     
     
         29 . The method of  claim 28 , wherein the vaccine comprises at least one polypeptide selected from the group consisting of BZLF 1, BRLF 1, and BMLF 1 and wherein the replicative phase is induced by providing to said cell at least one polypeptide selected from the group consisting of BZLF 1, BRLF 1, and BMLF 1. 
     
     
         30 . The method of  claim 29 , wherein the polypeptide is BZLF1. 
     
     
         31 . The method of  claim 29 , wherein the cells are transfected with a stable vector. 
     
     
         32 . The method of  claim 31 , wherein expression of any of said EBV polypeptides is inducibly regulated. 
     
     
         33 . A cell obtained by any of the steps of the method of  claim 29 . 
     
     
         34 . A kit comprising the vaccine of  claim 15  and instructions for use thereof. 
     
     
         35 . A method for generating CD8+ T cells specific for an EBV antigen comprising contacting a cell with the vaccine of  claim 15 .