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HBV VACCINE

20200113998 ยท 2020-04-16

    Inventors

    Cpc classification

    International classification

    Abstract

    The invention relates to a multi-HBV immunogen viral vector vaccine comprising: a viral vector comprising an immunogen expression cassette, wherein the expression of a protein encoded by the expression cassette is arranged to be driven by a promoter, wherein the immunogen expression cassette encodes: a) HBV Core; b) a modified HBV polymerase (P.sub.mut), wherein the modification is a mutation to wild-type HBV polymerase to substantially remove polymerase function; c) HBV surface antigen (HbsAg); and d) an intergenic sequence that is arranged to cause expression of at least the HBV surface antigen (HbsAg) as a separate protein from the HBV core and the modified HBV polymerase (P.sub.mut), wherein the intergenic sequence is downstream (3) of the sequences encoding the HBV core and the modified HBV polymerase (P.sub.mut) and upstream (5) of the sequence encoding the HBV surface antigen (HbsAg); and related compositions, vaccination methods and methods of treatment or prophylaxis of HBV infection.

    Claims

    1. A multi-HBV immunogen viral vector vaccine comprising: a viral vector comprising an immunogen expression cassette, wherein the expression of a protein encoded by the expression cassette is arranged to be driven by a promoter, wherein the immunogen expression cassette encodes: a) HBV Core; b) a modified HBV polymerase (P.sub.mut), wherein the modification is a mutation to wild-type HBV polymerase to substantially remove polymerase function; c) HBV surface antigen (HbsAg); and d) an intergenic sequence that is arranged to cause expression of at least the HBV surface antigen (HbsAg) as a separate protein from the HBV core and the modified HBV polymerase (P.sub.mut), wherein the intergenic sequence is downstream (3) of the sequences encoding the HBV core and the modified HBV polymerase (P.sub.mut) and upstream (5) of the sequence encoding the HBV surface antigen (HbsAg).

    2. The multi-HBV immunogen viral vector vaccine according to claim 1, wherein the intergenic sequence comprises a cleavage domain, an IRES (Internal Ribosomal Entry Site), a splicing signal, or a secondary promoter.

    3. The multi-HBV immunogen viral vector vaccine according to claim 1, wherein the intergenic sequence comprises a cleavage domain; optionally wherein the cleavage domain comprises a ribosome skipping cleavage domain; further optionally, wherein the cleavage domain comprises or consists of Furin-2A (F2A) peptide sequence or a functional variant thereof.

    4. The multi-HBV immunogen viral vector vaccine according to claim 1, wherein the intergenic sequence comprises a secondary promoter to promote expression of at least the surface antigen (HbsAg).

    5. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the immunogen expression cassette further encodes HBV Pre-Core (PreC).

    6. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the immunogen expression cassette further encodes HBV PreS1, and/or a truncated form thereof.

    7. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the immunogen expression cassette further encodes HBV PreS2.

    8. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the immunogen expression cassette encodes HBV Pre-Core (PreC) and HBV PreS1, and a truncated form of PreS1.

    9. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the immunogen expression cassette is capable of expressing HBV e-Antigen.

    10. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the HBV Core and modified polymerase (Pmut) are arranged to be expressed as a fusion protein.

    11. The multi-HBV immunogen viral vector vaccine according to claims 5 to 10, wherein the HBV Pre-core, HBV Core and modified polymerase (Pmut) are arranged to be expressed as a fusion protein.

    12. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the immunogen expression cassette does not encode HBV X protein.

    13. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the immunogen expression cassette comprises nucleic acid comprising the sequence of: SEQ ID NO: 46 (SIi-HBV-CPmutS) or a variant thereof; SEQ ID NO: 47 (SIi-HBV-SCPmut) or a variant thereof; SEQ ID NO: 48 (SIi-HBV-CPmutPreS-S(sh)) or a variant thereof; SEQ ID NO: 49 (SIi-HBV-CPmutPreS-TPA-S(sh)) or a variant thereof; SEQ ID NO: 24 (MVA-SIi-HBV-PreS-Pmut-C-S(sh)) or a variant thereof; or SEQ ID NO: 27 or SEQ ID NO: 58 (MVA-SIi-HBV-PreS-Pmut-C-TPA-S(sh)) or a variant thereof.

    14. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the viral vector encodes the amino acid sequence of: SEQ ID NO: 3 (SIi-HBV-CPmutS) or a variant thereof. SEQ ID NO: 11 (SIi-HBV-SCPmut) or a variant thereof. SEQ ID NO: 13 (SIi-HBV-CPmutPreS-S(sh)) or a variant thereof. SEQ ID NO: 25 (SIi-HBV-CPmutPreS-TPA-S(sh)) or a variant thereof. SEQ ID NO: 23 (MVA-SIi-HBV-PreS-Pmut-C-S(sh)) or a variant thereof. SEQ ID NO: 26 (MVA-SIi-HBV-PreS-Pmut-C-TPA-S(sh)) or a variant thereof.

    15. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the viral vector comprises an adenovirus vector or Modified Vaccinia Ankara (MVA) vector.

    16. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the viral vector may comprise a group E simian adenovirus vector.

    17. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the promoter is encoded in the immunogen expression cassette, for example the promoter may be encoded at, or adjacent to, the 5 end of the immunogen expression cassette; or wherein the promoter may be encoded as part of the viral vector nucleic acid outside of the immunogen expression cassette.

    18. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the promoter promotes the expression of all the encoded protein of the immunogen expression cassette.

    19. The multi-HBV immunogen viral vector vaccine according to claims 1 to 17, wherein the immunogen expression cassette comprises a secondary promoter, wherein the promoter is a primary promoter that is arranged to promote expression of at least the HBV core and modified polymerase (Pmut), and not the HBV surface antigen (HbsAg) which is arranged to be promoted separately by the secondary promoter.

    20. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the promoter comprises a CMV promoter or a pox viral promoter.

    21. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the HBV Core comprises or consists of a full length wild-type HBV Core sequence.

    22. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the modified HBV polymerase (P.sub.mut) is not a truncated form of HBV polymerase (i.e. it is full length relative to wildtype HBV polymerase).

    23. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the modified HBV polymerase (P.sub.mut) comprises or consists of the sequence of SEQ ID NO: 8 or a variant thereof.

    24. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the HbsAg comprises or consists of a full length wild-type HbsAg sequence, or a variant thereof.

    25. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the immunogen expression cassette encodes a truncated form of HBV PreS1 and the truncated PreS1 is arranged to be expressed a fusion protein with the HBV surface antigen (S/HbsAg).

    26. The multi-HBV immunogen viral vector vaccine according to claim 25, further comprising a linker sequence provided between the truncated PreS1 and surface antigen (S/HbsAg).

    27. The multi-HBV immunogen viral vector vaccine according to claim 25 or 26, wherein the truncated PreS1 with a fused surface antigen is encoded downstream (3) of the intergenic sequence.

    28. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the expression cassette encodes a NPreS1 and PreS2 fusion sequence.

    29. The multi-HBV immunogen viral vector vaccine according to claim 28, wherein the encoded NPreS1 and PreS2 fusion sequence is encoded upstream (5) of the intergenic sequence.

    30. The multi-HBV immunogen viral vector vaccine according to claim 29, wherein the encoded NPreS1 and PreS2 fusion sequence is further fused with the modified polymerase (Pmut).

    31. The multi-HBV immunogen viral vector vaccine according to claim 30, wherein a linker sequence is provided between the PreS2 and modified polymerase (Pmut).

    32. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein the immunogen expression cassette further encodes a peptide adjuvant.

    33. The multi-HBV immunogen viral vector vaccine according to claim 32, wherein the peptide adjuvant comprises TPA (tissue plasminogen activator) or a human or non-human invariant chain (Ii), or a fragment thereof.

    34. The multi-HBV immunogen viral vector vaccine according to any preceding claim, wherein linker residues are encoded between one or more, or all, of the protein/antigen sequences that are provided in a fusion protein.

    35. A nucleic acid comprising or consisting of an HBV immunogen expression cassette, wherein the immunogen expression cassette encodes: a) HBV Core; b) a modified HBV polymerase (P.sub.mut), wherein the modification is a mutation to wild-type HBV polymerase to substantially remove polymerase function; c) HBV surface antigen (HbsAg); and d) an intergenic sequence that is arranged to cause expression of at least the HBV surface antigen (HbsAg) as a separate protein from the HBV core and the modified HBV polymerase (P.sub.mut), wherein the intergenic sequence is downstream (3) of the sequences encoding the HBV core and the modified HBV polymerase (P.sub.mut) and upstream (5) of the sequence encoding the HBV surface antigen (HbsAg).

    36. The nucleic acid according to claim 35, wherein the immunogen expression cassette further encodes a promoter.

    37. The nucleic acid according to claim 35 or 36, wherein the immunogen expression cassette is isolated or provided in a non-viral vector.

    38. A composition comprising the viral vector according to any of claims 1 to 34 or the nucleic acid according to any of claims 35 to 37, optionally wherein the composition is a pharmaceutically acceptable composition.

    39. The composition according to any of claim 38, further comprising another therapeutically or prophylactically active ingredient and/or an adjuvant.

    40. The composition according to claim 38 or 39, wherein the composition comprises a pharmaceutically acceptable carrier and/or wherein the composition is a vaccine composition.

    41. The composition according to any of claims 38 to 40, the viral vector according to any of claims 1 to 34 or the nucleic acid according to any of claims 35 to 37, for use in the prophylaxis or treatment of HBV infection in a subject; optionally wherein the use is as a vaccine.

    42. A method of treatment or prophylaxis of HBV infection comprising the administration of the composition according to any of claims 38 to 40, the viral vector according to any of claims 1 to 34 or the nucleic acid according to any of claims 35 to 37, to a subject.

    43. The composition for the use according to claim 41 of the method according to claim 42, wherein the use or administration is in combination with the use or administration of another therapeutically or prophylactically active ingredient.

    44. A prime boost vaccination kit comprising a prime vaccination comprising the composition according to any of claims 38 to 40, the viral vector according to any of claims 1 to 34 or the nucleic acid according to any of claims 35 to 37; and a boost vaccination comprising the composition according to any of claims 38 to 40, the viral vector according to any of claims 1 to 34 or the nucleic acid according to any of claims 35 to 37.

    45. The prime and boost vaccination kit according to claim 44, wherein the prime and boost vaccinations comprise different viral vectors.

    Description

    [0175] Embodiments of the invention will now be described in more detail, by way of example only, with reference to the accompanying drawings.

    [0176] FIG. 1: (A) Phlyogenitic relationship of 1447 HBV genotype C sequences used to generate consensus sequence (B) Comparison of HBV genotype C consensus (SEQ ID NO: 1) and KP017269.1 HBV isolate JP-02 (SEQ ID NO: 2). 1447 HBV genotype C nucleotide sequences, downloaded from HBV data base, HBVdb: https://hbvdb.ibcp.fr/HBVdb/HBVdbIndex, were aligned using MAFFT (a multiple sequence alignment program), to generate the HBV genotype C consensus sequence and a phylogenetic tree and. Alignment of 3215 nucleotide sequences of the consensus and chosen patient's sequence (KP017269.1 HBV isolate JP-02) showed three nucleotide differences (at positions 52, 1053 and 2699), which are highlighted in grey colour. The sequence of the HBV genotype C consensus sequence generated from 1447 genotype c isolate jp-02 is provided below.

    [0177] FIG. 2: (A) HBV viral genome and codon layout. The HBV virion has a partial double stranded DNA (having a full length negative strand DNA and a partially synthesised positive strand DNA attached to the polymerase protein). The genome is approximately 3.2 Kb in length and has four major codons: core (including the precore region), polymerase, surface (including the preS1 and preS2 region) and X. (B) HBV Immunogen Layout. Two immunogens SIi-HBV-CPmutS and SIi-HBV-SCPmut were designed. Both immunogens encodes HBV codons (encompassing precore, core, polymerase [Pmut], preS1, preS2 and surface proteins) and non HBV codons (comprising of a truncated Shark Invariant chain [SIi], two linkers [represented in Turquoise blue colour] and a Furin 2A [F2A] peptide sequence). Within the mammalian expression cassette, the immunogen codon sequences are placed next to CMV promoter. (C) Invitro expression analysis. Plasmids encoding SIi-HBV-CPmutS and SIi-HBV-SCPmut were transfected into HEK293A cells. 24 hours post-transfection, cells were lysed and the lysates were analysed in Western blot experiments using mouse anti-HBV-PreS1 and mouse anti-HBV-Polymerase antibodies. Blots probed with mouse anti-GAPDH served as loading controls. (D) ChAdOX2-SIi-HBV-CPmutS. The mammalian expression cassette with SIi-HBV-CPmutS immunogen codons was inserted into the replication deficient ChAdOx2 vector. Recombinant ChAdOX2-SIi-HBV-CPmutS virus was generated by transfecting the ChAdOX2-SIi-HBV-CPmutS vector into T-REx-293 cells (Thermo Fisher Scientific) using standard methods as previously described.

    [0178] FIG. 3: Testing ChAdOx2-SIi-HBV-CPmutS vaccine in naive mice models. (A) Spleenocyte responses in BALB/c mice (B) Intra Hepatic

    [0179] Lymphocyte responses in BALB/c mice (C) Spleenocyte responses in CD1 mice (D) Intra Hepatic Lymphocyte responses in CD1 mice. BALB/c mice (4 mice at the age of 7 weeks) and CD1 (2 mice at age of 18 weeks and 5 mice at age of 13 weeks) were vaccinated by intramuscular injections with 410.sup.7 IU and 510.sup.7 IU per mice respectively of ChAdOx2-SIi-HBV-CPmutS vaccine. 14 days post-vaccination, mice were sacrificed, spleenocytes and intra hepatic lymphocytes (IHL) were isolated from spleen and liver according to standard protocol. 210.sup.5 spleenocytes and 110.sup.5 IHL's were plated on to ELSIPOT plates (pre-coated with Anti-mouse INFy monoclonal antibody, by overnight incubation) along with DMSO (1%) or non-HBV peptide pool (A,I, L at a concentration of 3 g/ml) or HBV peptide pool (Core, Pol-1, Pol-2, Pol-3, Pol-4, PreS1/S2 and Surface at a concentration of 3 g/ml) or a positive control mitogen (PHA or Concanavalin A at a concentration of 10 /ml and 12.5 /ml respectively). After an overnight 15 hours incubation at 37 C., the plates were washed 7 with PBS and incubated with Biotin conjugated mouse anti-INF for 2 hours at room temperature, followed by 4 wash with PBS and AP conjugated anti-biotin incubation for 2 hours at room temperature. The plates were then washed 4 with PBS and developed with BCIP/NBT substrate until spots appeared on the wells. After a final wash with water and drying the spot forming units (SFU) per million cells from individual wells were counted on a automated ELISpot plate reader. ELISPOT responses for each peptide pool stimulus with spleenocytes and IHL's from BALB/c and CD1 mice are represented in separate charts. Total responses from HBV peptide stimulus were also calculated by combining ELISPOT responses to all HBV peptide pools (Core, Pol-1, Pol-2, Pol-3, Pol-4, PreS1/S2 and Surface).

    [0180] FIG. 4: Comparison of the immunogenicity of HBV immunogen SIi-HBV-CPmutS with and without Shark Invariant chain (SIi). (A) HBV-CPmutS immunogen layout. The immunogens HBV-CPmutS was generated by deleting the SIi and first linker present in SIi-HBV-CPmutS. HBV-CPmutS has exactly the same amino acids, except that it lacks the SIi and first linker sequence that is present in the SIi-HBV-CPmutS immunogen. (B) Spleenocyte responses in CD1 mice. (C) Intra Hepatic Lymphocyte responses in CD1 mice. ChAdOX2-HBV-CPmutS was generated as described earlier and CD1 mice (10 mice at age of 7 weeks) were vaccinated by intramuscular injections with 510.sup.7 IU per mice ChAdOx2-HBV-CPmutS vaccine and 14 days post-vaccination mice were sacrificed and spleenocytes and intra hepatic lymphocytes response data were generated, as described earlier. Data from 5 and 10 CD1 mice vaccinated with ChAdOx2-SIi-HBV-CPmutS and ChAdOx2-HBV-CPmutS respectively were compared and represented in charts for ease of comparision.

    [0181] FIG. 5A: Schematic layout of SIi-HBV-CPmutPreS-S(sh).

    [0182] FIG. 5B: Schematic layout of SIi-HBV-CPmutPreS-TPA-S(sh).

    [0183] FIG. 6A: Schematic layout of MVA-SIi-HBV-PreS-P.sub.mut-C-S(sh).

    [0184] FIG. 6B: Schematic layout of MVA-SIi-HBV-PreS-Pmut-C-TPA-S(sh).

    [0185] FIG. 7: Testing ChAdOx1-SIi-HBV-CPmutS vaccine in BALB/c mice, splenocyte response: BALB/c mice (8 mice at the age of 8 weeks) were vaccinated by intramuscular injections with 510.sup.7 IU per mice of ChAdOx1-SIi-HBV-CPmutS vaccine. 14 days post-vaccination, mice were sacrificed; splenocyte response data were generated, as described earlier. Splenocyte T-cell response for each peptide pool stimulus and pooled total response from all HBV-peptide pools are represented in the bar chart.

    [0186] FIG. 8: Comparison of the immunogenicity of HBV immunogen SIi-HBV-CPmutS encoded via ChAdOx1 and ChAdOx2. HBV-CPmutS immunogen encoding ChAdOx1 and ChAdOx2 viral vectored vaccines were generated, as previously described (FIG. 2D). 10 CD1 mice at age of 7 weeks and 8 CD1 mice at age of 8 weeks were vaccinated by intramuscular injections with 510.sup.7 IU per mice of ChAdOx2-HBV-CPmutS and ChAdOx1-HBV-CPmutS vaccine, respectively, and 14 days post-vaccination mice were sacrificed and splenocyte response data were generated, as described earlier. Data from 10 and 8 CD1 mice vaccinated with ChAdOx2-SIi-HBV-CPmutS and ChAdOx1-HBV-CPmutS were compared and represented in charts for ease of comparison. A statistically significant higher magnitude of total splenocyte T-cell response was observed in mice vaccinated with ChAdOx2-HBV-CPmutS vaccine, compared to the ChAdOx1-HBV-CPmutS vaccine.

    [0187] FIG. 9: Comparison of the immunogenicity of HBV immunogen SIi-HBV-CPmut-PreS-S(sh) (termed as HBV-v2) encoded via ChAdOx1 and ChAdOx2 viral vectors using either a short CMV promoter or long CMV promoter. Two short CMV promoter and one long CMV promoter based SIi-HBV-CPmutPreS-S(sh) immunogen (termed as HBV-v2) encoding ChAdOx1 (ChAdOx1-LP-HBV-v2 and ChAdOx1-SP-HBV-v2) and ChAdOx2 (ChAdOx2-SP-HBV-v2) vaccines were generated as previously described. 15 Balbc mice at age of 8 weeks were split into 3 groups (5 mice per group) and vaccinated by intramuscular injections with 510.sup.7 1U of ChAdOx1-SP-HBV-v2 or ChAdOx2-SP-HBV-v2 or ChAdOx1-LP-HBV-v2 vaccine. 14 days post-vaccination mice were sacrificed and splenocyte response data were generated as previously described. Comparative data from all three HBV-v2 vaccines are represented in the same chart for ease of comparison. A statistically significant higher magnitude of total splenocyte T-cell response was observed in mice vaccinated with ChAdOx1-SP-HBV-v2 vaccine, compared to the other two HBV-v2 vaccines. In addition, the ChAdOx2-SP-HBV-v2 vaccine showed a statistically significant higher magnitude of total splenocyte T-cell response compared to the ChAdOx1-LP-HBV-v2 vaccine.

    [0188] FIG. 10A-G: show immunogen layout for (A) SIi-HBV-CPmutS; (B) SIi-HBV-SCP.sub.mut (C) HBV-CP.sub.mutS (D) SIi-HBV-CP.sub.mutPreS-S(sh) (E) MVA-SIi-HBV-PreS-P.sub.mut-C-S(sh) (F) SIi-HBV-CP.sub.mutPreS-TPA-S(sh), and (G) MVA-SIi-HBV-PreS-P.sub.mut-C-TPA-S(sh).

    [0189] A HBV vaccine was generated based on the HBV genotype C, one of the commonest HBV genotypes in South East Asia that has more frequent association with chronic HBV infection. To design the HBV immunogen, a patient's HBV genotype C sequence (GeneBank: KP017269.1) was selected that was closest to the consensus, this was generated by aligning 1447 HBV-genotype-C sequences from HBVdb, a Hepatitis Virus B database (FIGS. 1A and 1B). The chosen HBV genotype C sequence had only three nucleotide changes compared to the consensus, of which two were silent mutations and one had a mutation in the polymerase protein (FIG. 1C).

    [0190] A strong and multi-antigen specific T cell response against different HBV proteins is believed to play a major role in viral clearance of a resolving HBV infection. Based on this HBV immunogens have been designed that encode all major proteins of the virus, namely the core (including the Pre-core region), a non-functional polymerase Pmut (Pmut: HBV polymerase with functional mutations, aiming to discourage the vaccine encoded polymerase's ability to participate in HBV viral replication) and the surface protein (including its PreS1 and PreS2 regions). FIG. 2A and 2B shows the codon layout of HBV genome and a schematic representation of the first two HBV vaccine immunogen designs respectively. The layout has been designed to encode pre-core, core and Pmut as a fusion protein and a separate surface protein using a furin 2A (F2A) peptide cleavage mechanism, which by causing ribosomal skipping events, helps to encode two proteins from a single open reading frame.

    [0191] Encoding multiple proteins within a single transgene cassette requires careful design, where the proximity of codon sequence to CMV promoter plays an important role in the level of expression of the encoded proteins. To analyse this two immunogens (FIG. 2B), SIi-HBV-CPmutS and SIi-HBV-SCPmut, we generated which encodes the surface protein either at the end or the beginning of the immunogen cassette respectively and tested them in Western blot expression studies. Western blot expression analysis showed that layout SIi-HBV-CPmutS has the best F2A cleavage possibility, which generates large amounts of the expected CPmut and S proteins when compared to the SIi-HBV-SCPmut (FIG. 2c).

    [0192] Based on this observation, we decided to take forward SIi-HBV-CPmutS immunogen layout for the generation of chimpanzee adenovirus (ChAdOx2) based T-cell inducing HBV vaccine (FIG. 2D). A small scale batch of ChAdOx2-SIi-HBV-CPmutS was manufactured at the viral vector core facility of the Jenner Institute, University of Oxford, following good manufacturing practices, and used in mice immunogenicity studies.

    [0193] The ability of ChAdOx2-SIi-HBV-CPmutS vaccine to generate a T-cell immune response was tested using naive mice models. Naive BALB/c and CD1 mice were immunized with 410.sup.7 IU and 510.sup.7 IU per mice respectively of ChAdOx2-SIi-HBV-CPmutS vaccine by intra-muscular injections. 14 days post vaccination mice were sacrificed and IFN- ELISPOT assays were performed with spleenocytes and intra hepatic lymphocytes (IHL) isolated from spleen and liver. Synthetic peptides of 15mers, overlapping by 11 amino acids, generated across the whole of SIi-HBV-CPmutS immunogen, were combined into specific pools (representing different regions of the SIi-HBV-CPmutS immunogen) and used as a stimulant in IFN- ELISPOT assays. Results showed that the ChAdOx2-SIi-HBV-CPmutS vaccine generated a good IFN- ELISPOT response to SIi-HBV-CPmutS immunogen (FIG. 3). Both BALB/c and CD1 mice showed stronger responses to the polymerase and surface proteins, a weak response to the core protein and negligible or no response to the non-HBV proteins (Shark Invariant chain [SIi], F2A and linkers) in the immunogen.

    [0194] Previous studies have shown that the shark invariant chain (SIi), when placed at the N-terminus of the immunogen, functions as a molecular adjuvant and increases the overall T-cell immune response to the immunogen. To test this, we generated a HBV immunogen without SIi (HBV-CPmutS vaccine) (FIG. 4A), and then generated a ChAdOx2-HBV-CPmutS vaccine and tested them in similar CD1 mice immunogenicity experiments. Results showed that the SIi vaccine (ChAdOx2-Sli-HBV-CPmutS) generates a higher magnitude of IFN- ELISPOT response (both Splenocyte and Intra Hepatic Lymphocyte IFN- ELISPOT response, represented in FIGS. 4B, 4C) compared to the Non-SIi vaccine (ChAdOx2-HBV-CPmutS).

    [0195] The main aim of the viral vectored HBV vaccine design is to generate both T-cell and antibody response to the HBV immunogen. In order to generate a successful antibody response, the immunogen encoded protein has to fold into its native conformation. To provide this, HBV immunogens, SIi-HBV-CPmutPreS-S(sh) and SIi-HBV-CPmutPreS-TPA-S(sh) (FIG. 5A and 5B) were designed, which encode the N-terminal half of PreS1 domain fused with a linker to the S domain of surface protein, as the antibody inducing immunogen component, and the remaining peptide sequences of the surface protein (the C-terminal half of PreS1 domain and the whole PreS2 domain) were fused to the PreCore/Core/Pmut, in order to preserve the T-cell epitopes that would be required for generation of T-cell immune response. FIG. 5A and 5B shows a schematic layout of SIi-HBV-CPmutPreS-S(sh).

    [0196] MVA-HBV immunogens were also designed, which followed designs similar to SIi-HBV-CPmutPreS-S(sh) and and SIi-HBV-CPmutPreS-TPA-S(sh). However, the T-cell component of HBV immunogen SIi-HBV-CPmutPreS is encoded by the early promoter F11 and the antibody inducing component S(sh) or TPA-S(sh) is encoded by the early/late promoter mH5. The cloning cassette also has F11-left flank and F11-right flank sequences, to allow insertion into the F 11 locus, by homologous recombination. FIG. 6A and 6B shows schematic layouts of MVA-SIi-HBV-PreS-Pmut-C-S(sh) and MVA-SIi-HBV-CPmutPreS-TPA-S(sh) respectively.

    [0197] Conclusion: The novel multi-antigen HBV immunogen based ChAdOx2-SIi-HBV-CPmutS are highly immunogenic in naive mice models. These studies pave way for future studies in chronic HBV mice models, to assess their ability to overcome chronic HBV infection through immunotherapeutic approaches.

    [0198] Sequences:

    [0199] HBV genotype C consensus sequence generated from 1447 genotype C sequences (3215 base pairs) is provided as SEQ ID NO: 1. The Sequence of KP017269.1 HBV isolate JP-02 is provided as SEQ ID NO: 2.

    [0200] Example Cassettes

    [0201] 1. SIi-HBV-CP.sub.mutS [0202] 1.1. SIi-HBV-CP.sub.mutS: Immunogen layout is shown in FIG. 10A

    TABLE-US-00002 1.2.SIi-HBV-CP.sub.mutS:Aminoacidsequence (SEQIDNO:3) MSLLWGGVTVLAAMLIAGQVASVVFLVKGGGPGGGMQLFHLCLIISCS CPTVQASKLCLGWLWGMDIDPYKEFGASVELLSFLPSDFFPSIRDLLD TASALYREALESPEHCSPHHTALRQAILCWGELMNLATWVGSNLEDPA SRELVVSYVNVNMGLKIRQLLWFHISCLTFGRETVLEYLVSFGVWIRT PPAYRPPNAPILSTLPETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRS QSRESQCGGGSGGGMPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLN RRVAEDLNLGNLNVSIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHI HLQEDIINRCQQYVGPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIK PYYPEHAVNHYFKTRHYLHTLWKAGILYKRETTRSASFCGSPYSWEQE LQHGRLVFQTSTRHGDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQ PQQGSLARGKSGRSGSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSS CLHQSAVRKTAYSHLSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFS AWWLQFRNSKPASDYALTHIVNLLEDWGPATEHGEHNIRIPRTPARVT GGVFLVDKNPHNTTESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNL LSSNLSWLSLDVSAAFYHIPLHPAAMPHLLVGSSGLPRYVARLSSTSR NINYQHGTMQDLHDSCSRNLYVSLLLLYKTFGRKLHLYSHPIILGFRK IPMGVGLSPFLLAQFTSAICSVVRRAFPHCLAFSYMDDVVLGAKSVQH LESLFTSITNFLLSLGIHLNPNKTKRWGYSLNFMGYVIGSWGTLPQEH IVLKIKQCFRKLPVNRPIDWKVCQRIVGLLGFAAPFTQCGYPALMPLY ACIQSKQAFTFSPTYKAFLCKQYLNLYPVARQRSGLCQVFADATPTGW GLAIGHRAMRGTFVAPLPIHTAELLAACFARSRSGAKLIGTDNSVVLS RKYTSFPWLLGCAANWILRGTSFVYVPSALNPAADPSAGRLGLYRPLL HLPFRPTTGRTSLYAVSPSVPSHLPDRVHFASPLHVAWRPPRKRRAPV KQTLNFDLLKLAGDVESNPGPMGGWSSKPRQGMGTNLSVPNPLGFFPD HQLDPAFGANSNNPDWDFNPNKDHWPEANQVGAGAFGPGFTPPHGGLL GWSPQAQGILTTVPAAPPPASTNRQSGRQPTPISPPLRDSHPQAMQWN STTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSRTGDP APNMENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGA PTCPGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIF LLVLLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTK PSDGNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPT VWLSVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI 1.3.SIi-HBV-CP.sub.mutS:Descriptionforaminoacid sequencesofHBVimmunogen 1.3.1.Firstaminoacidofpolypeptide=M 1.3.2.SharkInvariantchain(SIi)= (SEQIDNO:4) SLLWGGVTVLAAMLIAGQVASVVFLV 1.3.3.Linker= (SEQIDNO:5) KGGGPGGG 1.3.4.C= 1.3.4.1.PreC= (SEQIDNO:16) MQLFHLCLIISCSCPTVQASKLCLGWLWG 1.3.4.2.Core= (SEQIDNO:6) MDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYREALESPEHC SPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSYVNVNMGLK IRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPNAPILSTLP ETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQC 1.3.5.Linker= (SEQIDNO:7) GGGSGGG 1.3.6.Pmut=(mutations:Y63A,C323A,C334A, C338A,C352A,R714A,D788A,R792A) (SEQIDNO:8) MPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLNRRVAEDLNLGNLNV SIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHIHLQEDIINRCQQYV GPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIKPYYPEHAVNHYFKT RHYLHTLWKAGILYKRETTRSASFCGSPYSWEQELQHGRLVFQTSTRH GDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQPQQGSLARGKSGRS GSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSSCLHQSAVRKTAYSH LSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFSAWWLQFRNSKPASD YALTHIVNLLEDWGPATEHGEHNIRIPRTPARVTGGVFLVDKNPHNTT ESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNLLSSNLSTFGRKLHL YSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVRRAFPHCLAFSYMD DVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKTKRWGYSLNFMGYV IGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQRIVGLLGFAAPFT QCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYLNLYPVARQRSGLC QVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAELLAACFARSRSGAK LIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFVYVPSALNPAADPS AGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHLPDRVHFASPLHVA WRPP 1.3.7.Furin2A(F2A)= (SEQIDNO:9) RKRRAPVKQTLNFDLLKLAGDVESNPGP 1.3.8.Surfaceproteins(S)= 1.3.8.1.PreS1= (SEQIDNO:52) MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGILTTVPAAPPPAS TNRQSGRQPTPISPPLRDSHPQA 1.3.8.2.PreS2= (SEQIDNO:53) MQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSR TGDPAPN 1.3.8.3.Surface(S)= (SEQIDNO:10) MENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTC PGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLV LLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSD GNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWL SVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI

    [0203] The SIi-HBV-CPmutS nucleotide sequence is provided as SEQ ID NO: 46.

    [0204] 2. SIi-HBV-SCP.sub.mut [0205] 2.1. SIi-HBV-SCP.sub.mut: Immunogen layout is shown in FIG. 10B

    TABLE-US-00003 2.2.SIi_HBV-SCP.sub.mut:Aminoacidsequence (SEQIDNO:11) MSLLWGGVTVLAAMLIAGQVASVVFLVKGGSMGGWSSKPRQGMGTNLS VPNPLGFFPDHQLDPAFGANSNNPDWDFNPNKDHWPEANQVGAGAFGP GFTPPHGGLLGWSPQAQGILTTVPAAPPPASTNRQSGRQPTPISPPLR DSHPQAMQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTASPI SSIFSRTGDPAPNMENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSW WTSLNFLGGAPTCPGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIF LFILLLCLIFLLVLLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGT SMFPSCCCTKPSDGNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPF VQWFVGLSPTVWLSVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYIR KRRAPVKQTLNFDLLKLAGDVESNPGPMQLFHLCLIISCSCPTVQASK LCLGWLWGMDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYRE ALESPEHCSPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSY VNVNMGLKIRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPN APILSTLPETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQCG GGSGGGMPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLNRRVAEDLN LGNLNVSIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHIHLQEDIIN RCQQYVGPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIKPYYPEHAV NHYFKTRHYLHTLWKAGILYKRETTRSASFCGSPYSWEQELQHGRLVF QTSTRHGDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQPQQGSLAR GKSGRSGSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSSCLHQSAVR KTAYSHLSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFSAWWLQFRN SKPASDYALTHIVNLLEDWGPATEHGEHNIRIPRTPARVTGGVFLVDK NPHNTTESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNLLSSNLSWL SLDVSAAFYHIPLHPAAMPHLLVGSSGLPRYVARLSSTSRNINYQHGT MQDLHDSCSRNLYVSLLLLYKTFGRKLHLYSHPIILGFRKIPMGVGLS PFLLAQFTSAICSVVRRAFPHCLAFSYMDDVVLGAKSVQHLESLFTSI TNFLLSLGIHLNPNKTKRWGYSLNFMGYVIGSWGTLPQEHIVLKIKQC FRKLPVNRPIDWKVCQRIVGLLGFAAPFTQCGYPALMPLYACIQSKQA FTFSPTYKAFLCKQYLNLYPVARQRSGLCQVFADATPTGWGLAIGHRA MRGTFVAPLPIHTAELLAACFARSRSGAKLIGTDNSVVLSRKYTSFPW LLGCAANWILRGTSFVYVPSALNPAADPSAGRLGLYRPLLHLPFRPTT GRTSLYAVSPSVPSHLPDRVHFASPLHVAWRPP 2.3.SIi-HBV-SCP.sub.mut:Descriptionforaminoacid sequencesofHBVimmunogen 2.3.1.Firstaminoacidofpolypeptide=M 2.3.2.SharkInvariantchain(SIi)= (SEQIDNO:4) SLLWGGVTVLAAMLIAGQVASVVFLV 2.3.3.Linker= (SEQIDNO:14) KGGS 2.3.4.Surfaceproteins(S)= 2.3.4.1.PreS1= (SEQIDNO:52) MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGILTTVPAAPPPAS TNRQSGRQPTPISPPLRDSHPQA 2.3.4.2.PreS2= (SEQIDNO:53) MQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSR TGDPAPN 2.3.4.3.Surface(S)= (SEQIDNO:10) MENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTC PGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLV LLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSD GNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWL SVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI 2.3.5.Furin2A(F2A)= (SEQIDNO:9) RKRRAPVKQTLNFDLLKLAGDVESNPGP 2.3.6.C= 2.3.6.1.PreCore= (SEQIDNO:16) MQLFHLCLIISCSCPTVQASKLCLGWLWG 2.3.6.2.Core= (SEQIDNO:6) MDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYREALESPEHC SPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSYVNVNMGLK IRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPNAPILSTLP ETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQC 2.3.7.Linker= (SEQIDNO:7) GGGSGGG 2.3.8.Pmut=(mutations:Y63A,C323A,C334A, C338A,C352A,R714A,D788A,R792A) (SEQIDNO:8) MPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLNRRVAEDLNLGNLNV SIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHIHLQEDIINRCQQYV GPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIKPYYPEHAVNHYFKT RHYLHTLWKAGILYKRETTRSASFCGSPYSWEQELQHGRLVFQTSTRH GDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQPQQGSLARGKSGRS GSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSSCLHQSAVRKTAYSH LSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFSAWWLQFRNSKPASD YALTHIVNLLEDWGPATEHGEHNIRIPRTPARVTGGVFLVDKNPHNTT ESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNLLSSNLSTFGRKLHL YSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVRRAFPHCLAFSYMD DVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKTKRWGYSLNFMGYV IGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQRIVGLLGFAAPFT QCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYLNLYPVARQRSGLC QVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAELLAACFARSRSGAK LIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFVYVPSALNPAADPS AGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHLPDRVHFASPLHVA WRPP

    [0206] The SIi-HBV-SCPmut nucleotide sequence is provided as SEQ ID NO: 47:

    [0207] 3. HBV-CP.sub.mutS [0208] 3.1. HBV-CP.sub.mutS: Immunogen layout is shown in FIG. 10C

    TABLE-US-00004 3.2.HBV-CP.sub.mutS:Aminoacidsequence (SEQIDNO:12) MQLFHLCLIISCSCPTVQASKLCLGWLWGMDIDPYKEFGASVELLSFL PSDFFPSIRDLLDTASALYREALESPEHCSPHHTALRQAILCWGELMN LATWVGSNLEDPASRELVVSYVNVNMGLKIRQLLWFHISCLTFGRETV LEYLVSFGVWIRTPPAYRPPNAPILSTLPETTVVRRRGRSPRRRTPSP RRRRSQSPRRRRSQSRESQCGGGSGGGMPLSYQHFRKLLLLDDEAGPL EEELPRLADEGLNRRVAEDLNLGNLNVSIPWTHKVGNFTGLASSTVPV FNPEWQTPSFPHIHLQEDIINRCQQYVGPLTVNEKRRLKLIMPARFYP NLTKYLPLDKGIKPYYPEHAVNHYFKTRHYLHTLWKAGILYKRETTRS ASFCGSPYSWEQELQHGRLVFQTSTRHGDESFCSQSSGILSRSPVGPC VRSQLKQSRLGLQPQQGSLARGKSGRSGSIRARVHPTTRRSFGVEPSG SGHIDNSASSTSSCLHQSAVRKTAYSHLSTSKRQSSSGHAVELHNIPP SSARSQSEGPIFSAWWLQFRNSKPASDYALTHIVNLLEDWGPATEHGE HNIRIPRTPARVTGGVFLVDKNPHNTTESRLVVDFSQFSRGSTHVSWP KFAVPNLQSLTNLLSSNLSWLSLDVSAAFYHIPLHPAAMPHLLVGSSG LPRYVARLSSTSRNINYQHGTMQDLHDSCSRNLYVSLLLLYKTFGRKL HLYSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVRRAFPHCLAFSY MDDVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKTKRWGYSLNFMG YVIGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQRIVGLLGFAAP FTQCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYLNLYPVARQRSG LCQVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAELLAACFARSRSG AKLIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFVYVPSALNPAAD PSAGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHLPDRVHFASPLH VAWRPPRKRRAPVKQTLNFDLLKLAGDVESNPGPMGGWSSKPRQGMGT NLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPNKDHWPEANQVGAGA FGPGFTPPHGGLLGWSPQAQGILTTVPAAPPPASTNRQSGRQPTPISP PLRDSHPQAMQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTA SPISSIFSRTGDPAPNMENTTSGFLGPLLVLQAGFFLLTRILTIPQSL DSWWTSLNFLGGAPTCPGQNSQSPTSNHSPTSCPPICPGYRWMCLRRF IIFLFILLLCLIFLLVLLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPA QGTSMFPSCCCTKPSDGNCTCIPIPSSWAFARFLWEWASVRFSWLSLL VPFVQWFVGLSPTVWLSVIWMMWYWGPSLYNILSPFLPLLPIFFCLWV YI 3.3.HBV-CP.sub.mutS:Descriptionforaminoacid sequencesofHBVimmunogen 3.3.1.C= 3.3.1.1.PreCore= (SEQIDNO:16) MQLFHLCLIISCSCPTVQASKLCLGWLWG 3.3.1.2.Core= (SEQIDNO:6) MDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYREALESPEHC SPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSYVNVNMGLK IRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPNAPILSTLP ETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQC 3.3.2.Linker= (SEQIDNO:7) GGGSGGG 3.3.3.Pmut=(mutations:Y63A,C323A,C334A, C338A,C352A,R714A,D788A,R792A) (SEQIDNO:8) MPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLNRRVAEDLNLGNLNV SIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHIHLQEDIINRCQQYV GPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIKPYYPEHAVNHYFKT RHYLHTLWKAGILYKRETTRSASFCGSPYSWEQELQHGRLVFQTSTRH GDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQPQQGSLARGKSGRS GSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSSCLHQSAVRKTAYSH LSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFSAWWLQFRNSKPASD YALTHIVNLLEDWGPATEHGEHNIRIPRTPARVTGGVFLVDKNPHNTT ESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNLLSSNLSTFGRKLHL YSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVRRAFPHCLAFSYMD DVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKTKRWGYSLNFMGYV IGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQRIVGLLGFAAPFT QCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYLNLYPVARQRSGLC QVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAELLAACFARSRSGAK LIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFVYVPSALNPAADPS AGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHLPDRVHFASPLHVA WRPP 3.3.4.Furin2A(F2A)= (SEQIDNO:9) RKRRAPVKQTLNFDLLKLAGDVESNPGP 3.3.5.Surfaceproteins(S)= 3.3.5.1.PreS1= (SEQIDNO:52) MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGILTTVPAAPPPAS TNRQSGRQPTPISPPLRDSHPQA 3.3.5.2.PreS2= (SEQIDNO:53) MQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSR TGDPAPN 3.3.5.3.Surface(S) (SEQIDNO:10) MENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTC PGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLV LLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSD GNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWL SVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI 4.SIi-HBV-CP.sub.mutPreS-S(sh) 4.1.SIi-HBV-CP.sub.mutPreS-S(sh):Immunogenlayoutis showninFIG.10D 4.2.SIi-HBV-CP.sub.mutPreS-S(sh):Aminoacidsequence (SEQIDNO:13) MSLLWGGVTVLAAMLIAGQVASVVFLVKGGGPGGGMQLFHLCLIISCS CPTVQASKLCLGWLWGMDIDPYKEFGASVELLSFLPSDFFPSIRDLLD TASALYREALESPEHCSPHHTALRQAILCWGELMNLATWVGSNLEDPA SRELVVSYVNVNMGLKIRQLLWFHISCLTFGRETVLEYLVSFGVWIRT PPAYRPPNAPILSTLPETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRS QSRESQCGGGSGGGMPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLN RRVAEDLNLGNLNVSIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHI HLQEDIINRCQQYVGPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIK PYYPEHAVNHYFKTRHYLHTLWKAGILYKRETTRSASFCGSPYSWEQE LQHGRLVFQTSTRHGDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQ PQQGSLARGKSGRSGSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSS CLHQSAVRKTAYSHLSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFS AWWLQFRNSKPASDYALTHIVNLLEDWGPATEHGEHNIRIPRTPARVT GGVFLVDKNPHNTTESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNL LSSNLSTFGRKLHLYSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVV RRAFPHCLAFSYMDDVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNK TKRWGYSLNFMGYVIGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVC QRIVGLLGFAAPFTQCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQY LNLYPVARQRSGLCQVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAE LLAACFARSRSGAKLIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSF VYVPSALNPAADPSAGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSH LPDRVHFASPLHVAWRPPKSPNSNNPDWDFNPNKDHWPEANQVGAGAF GPGFTPPHGGLLGWSPQAQGILTTVPAAPPPASTNRQSGRQPTPISPP LRDSHPQAMQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTAS PISSIFSRTGDPAPNGSKGKRKRRAPVKQTLNFDLLKLAGDVESNPGP MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVGLEGGSGGMENTTSGFLGPLLVLQAGFFLLTRILTIPQ SLDSWWTSLNFLGGAPTCPGQNSQSPTSNHSPTSCPPICPGYRWMCLR RFIIFLFILLLCLIFLLVLLDYQGMLPVCPLLPGTSTTSTGPCKTCTI PAQGTSMFPSCCCTKPSDGNCTCIPIPSSWAFARFLWEWASVRFSWLS LLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYNILSPFLPLLPIFFCL WVYI 4.3.SIi-HBV-CP.sub.mutPreS-S(sh):Descriptionfor aminoacidsequencesofHBVimmunogen 4.3.1.Firstaminoacidofpolypeptide=M 4.3.2.SharkInvariantchain(SIi)= (SEQIDNO:4) SLLWGGVTVLAAMLIAGQVASVVFLV 4.3.3.Linker= (SEQIDNO:5) KGGGPGGG 4.3.4.C= 4.3.4.1.PreCore= (SEQIDNO:16) MQLFHLCLIISCSCPTVQASKLCLGWLWG 4.3.4.2.Core= (SEQIDNO:6) MDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYREALESPEHC SPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSYVNVNMGLK IRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPNAPILSTLP ETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQC 4.3.5.Linker= (SEQIDNO:7) GGGSGGG 4.3.6.Pmut=(mutations:Y63A,C323A,C334A, C338A,C352A,R714A,D788A,R792A) (SEQIDNO:8) MPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLNRRVAEDLNLGNLNV SIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHIHLQEDIINRCQQYV GPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIKPYYPEHAVNHYFKT RHYLHTLWKAGILYKRETTRSASFCGSPYSWEQELQHGRLVFQTSTRH GDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQPQQGSLARGKSGRS GSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSSCLHQSAVRKTAYSH LSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFSAWWLQFRNSKPASD YALTHIVNLLEDWGPATEHGEHNIRIPRTPARVTGGVFLVDKNPHNTT ESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNLLSSNLSTFGRKLHL YSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVRRAFPHCLAFSYMD DVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKTKRWGYSLNFMGYV IGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQRIVGLLGFAAPFT QCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYLNLYPVARQRSGLC QVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAELLAACFARSRSGAK LIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFVYVPSALNPAADPS AGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHLPDRVHFASPLHVA WRPP 4.3.7.Linker= KSP 4.3.8.NPreS1andPreS2= (SEQIDNO:15) NSNNPDWDFNPNKDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGI LTTVPAAPPPASTNRQSGRQPTPISPPLRDSHPQAMQWNSTTFHQALL DPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSRTGDPAPN 4.3.9.Linker= (SEQIDNO:20) GSKGK 4.3.10.Furin2A(F2A)= (SEQIDNO:9) RKRRAPVKQTLNFDLLKLAGDVESNPGP 4.3.11.S(sh)= 4.3.11.1.CPreS1= (SEQIDNO:21) MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVG 4.3.11.2.Linker= (SEQIDNO:22) LEGGSGG 4.3.11.3.Surface(S)= (SEQIDNO:18) MENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTC PGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLV LLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSD GNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWL SVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI

    [0209] The SIi-HBV-CPmutPreS-S(sh) nucleotide sequence is provided as SEQ ID NO: 48.

    [0210] 5. MVA-SIi-HBV-PreS-P.sub.mut-C-S(sh) [0211] 5.1. MVA-SIi-HBV-PreS-P.sub.mut-C-S(sh): Immunogen layout is shown in FIG. 10E

    TABLE-US-00005 5.2.MVA-SIi-HBV-PreS-P.sub.mut-C-S(sh):Aminoacid sequence (SEQIDNO:23) MSLLWGGVTVLAAMLIAGQVASVVFLVSKSGPPSGKSNSNNPDWDFNP NKDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGILTTVPAAPPPA STNRQSGRQPTPISPPLRDSHPQAMQWNSTTFHQALLDPRVRGLYFPA GGSSSGTVNPVPTTASPISSIFSRTGDPAPNGSKSGSKMPLSYQHFRK LLLLDDEAGPLEEELPRLADEGLNRRVAEDLNLGNLNVSIPWTHKVGN FTGLASSTVPVFNPEWQTPSFPHIHLQEDIINRCQQYVGPLTVNEKRR LKLIMPARFYPNLTKYLPLDKGIKPYYPEHAVNHYFKTRHYLHTLWKA GILYKRETTRSASFCGSPYSWEQELQHGRLVFQTSTRHGDESFCSQSS GILSRSPVGPCVRSQLKQSRLGLQPQQGSLARGKSGRSGSIRARVHPT TRRSFGVEPSGSGHIDNSASSTSSCLHQSAVRKTAYSHLSTSKRQSSS GHAVELHNIPPSSARSQSEGPIFSAWWLQFRNSKPASDYALTHIVNLL EDWGPATEHGEHNIRIPRTPARVTGGVFLVDKNPHNTTESRLVVDFSQ FSRGSTHVSWPKFAVPNLQSLTNLLSSNLSWLSLDVSAAFYHIPLHPA AMPHLLVGSSGLPRYVARLSSTSRNINYQHGTMQDLHDSCSRNLYVSL LLLYKTFGRKLHLYSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVR RAFPHCLAFSYMDDVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKT KRWGYSLNFMGYVIGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQ RIVGLLGFAAPFTQCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYL NLYPVARQRSGLCQVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAEL LAACFARSRSGAKLIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFV YVPSALNPAADPSAGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHL PDRVHFASPLHVAWRPPSKSPGSGPPMQLFHLCLIISCSCPTVQASKL CLGWLWGMDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYREA LESPEHCSPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSYV NVNMGLKIRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPNA PILSTLPETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQC MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVGASKGGKSGMENTTSGFLGPLLVLQAGFFLLTRILTIP QSLDSWWTSLNFLGGAPTCPGQNSQSPTSNHSPTSCPPICPGYRWMCL RRFIIFLFILLLCLIFLLVLLDYQGMLPVCPLLPGTSTTSTGPCKTCT IPAQGTSMFPSCCCTKPSDGNCTCIPIPSSWAFARFLWEWASVRFSWL SLLVPFVQWFVGLSPTVWLSVIWMMWYWGPSLYNILSPFLPLLPIFFC LWVYI 5.3.MVA-SIi-HBV-PreS-P.sub.mut-C-S(sh):Description foraminoacidsequencesofHBVimmunogen 5.3.1.1.Firstaminoacidofpolypeptide=M 5.3.1.2.SharkInvariantchain(SIi)= (SEQIDNO:4) SLLWGGVTVLAAMLIAGQVASVVFLV 5.3.1.3.Linker= (SEQIDNO:31) SKSGPPSGKS 5.3.1.4.NPreS1andPreS2= (SEQIDNO:15) NSNNPDWDFNPNKDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGI LTTVPAAPPPASTNRQSGRQPTPISPPLRDSHPQAMQWNSTTFHQALL DPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSRTGDPAPN 5.3.1.5.Linker= (SEQIDNO:32) GSKSGSK 5.3.1.6.Pmut=(mutations:Y63A,C323A,C334A, C338A,C352A,R714A,D788A,R792A) (SEQIDNO:8) MPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLNRRVAEDLNLGNLNV SIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHIHLQEDIINRCQQYV GPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIKPYYPEHAVNHYFKT RHYLHTLWKAGILYKRETTRSASFCGSPYSWEQELQHGRLVFQTSTRH GDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQPQQGSLARGKSGRS GSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSSCLHQSAVRKTAYSH LSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFSAWWLQFRNSKPASD YALTHIVNLLEDWGPATEHGEHNIRIPRTPARVTGGVFLVDKNPHNTT ESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNLLSSNLSTFGRKLHL YSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVRRAFPHCLAFSYMD DVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKTKRWGYSLNFMGYV IGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQRIVGLLGFAAPFT QCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYLNLYPVARQRSGLC QVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAELLAACFARSRSGAK LIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFVYVPSALNPAADPS AGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHLPDRVHFASPLHVA WRPP 5.3.1.7.Linker= (SEQIDNO:33) SKSPGSGPP 5.3.1.8.C= 5.3.1.8.1.PreCore (SEQIDNO:16) MQLFHLCLIISCSCPTVQASKLCLGWLWG 5.3.1.8.2.Core= (SEQIDNO:6) MDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYREALESPEHC SPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSYVNVNMGLK IRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPNAPILSTLP ETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQC 5.3.1.9.S(sh) 5.3.1.9.1.CPreS1= MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVG 5.3.1.9.2.Linker= (SEQIDNO:34) ASKGGKSG 5.3.1.9.3.Surface= (SEQIDNO:18) MENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTC PGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLV LLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSD GNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWL SVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI 5.4.NucleotidesequenceofMVA-SIi-HBV-PreS- P.sub.mut-C-S(sh)isprovidedasSEQIDNO:24. 5.5.Descriptionfornucleotidesequencesof MVA-SIi-HBV-PreS-P.sub.mut-C-S(sh): 5.5.1.F11-L-Flank=bases1-1097 (SEQIDNO:35) 5.5.2.SIi-HBV-PreS-P.sub.mut-C=bases1098-4838 (SEQIDNO:37) 5.5.3.Transcriptionterminatorsequence= bases4839-4845 TTTTTGT 5.5.4.mH5promoter=bases4846-4942 (SEQIDNO:28) 5.5.5.S(sh)=bases4943-5824(SEQIDNO:38) 5.5.6.F11-R-Flank=bases5825-7143 (SEQIDNO:36)

    [0212] 6. SIi-HBV-CP.sub.mutPreS-TPA-S(sh) [0213] 6.1. SIi-HBV-CP.sub.mutPreS-TPA-S(sh): Immunogen layout is shown in FIG. 10F

    TABLE-US-00006 6.2.SIi-HBV-CP.sub.mutPreS-TPA-S(sh):Aminoacid sequence (SEQIDNO:25) MSLLWGGVTVLAAMLIAGQVASVVFLVKGGGPGGGMQLFHLCLIISCS CPTVQASKLCLGWLWGMDIDPYKEFGASVELLSFLPSDFFPSIRDLLD TASALYREALESPEHCSPHHTALRQAILCWGELMNLATWVGSNLEDPA SRELVVSYVNVNMGLKIRQLLWFHISCLTFGRETVLEYLVSFGVWIRT PPAYRPPNAPILSTLPETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRS QSRESQCGGGSGGGMPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLN RRVAEDLNLGNLNVSIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHI HLQEDIINRCQQYVGPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIK PYYPEHAVNHYFKTRHYLHTLWKAGILYKRETTRSASFCGSPYSWEQE LQHGRLVFQTSTRHGDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQ PQQGSLARGKSGRSGSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSS CLHQSAVRKTAYSHLSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFS AWWLQFRNSKPASDYALTHIVNLLEDWGPATEHGEHNIRIPRTPARVT GGVFLVDKNPHNTTESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNL LSSNLSTFGRKLHLYSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVV RRAFPHCLAFSYMDDVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNK TKRWGYSLNFMGYVIGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVC QRIVGLLGFAAPFTQCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQY LNLYPVARQRSGLCQVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAE LLAACFARSRSGAKLIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSF VYVPSALNPAADPSAGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSH LPDRVHFASPLHVAWRPPKSPNSNNPDWDFNPNKDHWPEANQVGAGAF GPGFTPPHGGLLGWSPQAQGILTTVPAAPPPASTNRQSGRQPTPISPP LRDSHPQAMQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTAS PISSIFSRTGDPAPNGSKGKRKRRAPVKQTLNFDLLKLAGDVESNPGP MDAMKRGLCCVLLLCGAVFVSPSQEIHARFRRMGGWSSKPRQGMGTNL SVPNPLGFFPDHQLDPAFGANSNNPDWDFNPNKDHWPEANQVGLEGGS GGMENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAP TCPGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFL LVLLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKP SDGNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTV WLSVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI 6.3.SIi-HBV-CP.sub.mutPreS-TPA-S(sh):Descriptionfor aminoacidsequencesofHBVimmunogen 6.3.1.Firstaminoacidofpolypeptide=M 6.3.2.SharkInvariantchain(SIi)= (SEQIDNO:4) SLLWGGVTVLAAMLIAGQVASVVFLV 6.3.3.Linker= (SEQIDNO:5) KGGGPGGG 6.3.4.C= 6.3.4.1.PreCore (SEQIDNO:16) MQLFHLCLIISCSCPTVQASKLCLGWLWG 6.3.4.2.Core= (SEQIDNO:6) MDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYREALESPEHC SPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSYVNVNMGLK IRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPNAPILSTLP ETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQC 6.3.5.Linker= (SEQIDNO:7) GGGSGGG 6.3.6.Pmut=(mutations:Y63A,C323A,C334A, C338A,C352A,R714A,D788A,R792A) (SEQIDNO:8) MPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLNRRVAEDLNLGNLNV SIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHIHLQEDIINRCQQYV GPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIKPYYPEHAVNHYFKT RHYLHTLWKAGILYKRETTRSASFCGSPYSWEQELQHGRLVFQTSTRH GDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQPQQGSLARGKSGRS GSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSSCLHQSAVRKTAYSH LSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFSAWWLQFRNSKPASD YALTHIVNLLEDWGPATEHGEHNIRIPRTPARVTGGVFLVDKNPHNTT ESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNLLSSNLSTFGRKLHL YSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVRRAFPHCLAFSYMD DVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKTKRWGYSLNFMGYV IGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQRIVGLLGFAAPFT QCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYLNLYPVARQRSGLC QVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAELLAACFARSRSGAK LIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFVYVPSALNPAADPS AGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHLPDRVHFASPLHVA WRPP 6.3.7.Linker= KSP 6.3.8.NPreS1andPreS2= (SEQIDNO:15) NSNNPDWDFNPNKDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGI LTTVPAAPPPASTNRQSGRQPTPISPPLRDSHPQAMQWNSTTFHQALL DPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSRTGDPAPN 6.3.9.Linker= (SEQIDNO:20) GSKGK 6.3.10.Furin2A(F2A)= (SEQIDNO:9) RKRRAPVKQTLNFDLLKLAGDVESNPGP 6.3.11.TPA= (SEQIDNO:30) MDAMKRGLCCVLLLCGAVFVSPSQEIHARFRR 6.3.12.S(sh)= 6.3.12.1.CPreS1= (SEQIDNO:21) MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVG 6.3.12.2.Linker= (SEQIDNO:22) LEGGSGG 6.3.12.3.Surface(S)= (SEQIDNO:18) MENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTC PGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLV LLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSD GNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWL SVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI

    [0214] The SIi-HBV-CPmutPreS-TPA-S(sh) nucleotide sequence is provided as SEQ ID NO: 49.

    [0215] 7. MVA-SIi-HBV-PreS-P.sub.mut-C-TPA-S(sh) [0216] 7.1. MVA-SIi-HBV-PreS-P.sub.mut-C-TPA-S(sh): Immunogen layout is shown in FIG. 10G

    TABLE-US-00007 7.2.MVA-SIi-HBV-PreS-P.sub.mut-C-TPA-S(sh):Amino acidsequence (SEQIDNO:26) MSLLWGGVTVLAAMLIAGQVASVVFLVSKSGPPSGKSNSNNPDWDFNP NKDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGILTTVPAAPPPA STNRQSGRQPTPISPPLRDSHPQAMQWNSTTFHQALLDPRVRGLYFPA GGSSSGTVNPVPTTASPISSIFSRTGDPAPNGSKSGSKMPLSYQHFRK LLLLDDEAGPLEEELPRLADEGLNRRVAEDLNLGNLNVSIPWTHKVGN FTGLASSTVPVFNPEWQTPSFPHIHLQEDIINRCQQYVGPLTVNEKRR LKLIMPARFYPNLTKYLPLDKGIKPYYPEHAVNHYFKTRHYLHTLWKA GILYKRETTRSASFCGSPYSWEQELQHGRLVFQTSTRHGDESFCSQSS GILSRSPVGPCVRSQLKQSRLGLQPQQGSLARGKSGRSGSIRARVHPT TRRSFGVEPSGSGHIDNSASSTSSCLHQSAVRKTAYSHLSTSKRQSSS GHAVELHNIPPSSARSQSEGPIFSAWWLQFRNSKPASDYALTHIVNLL EDWGPATEHGEHNIRIPRTPARVTGGVFLVDKNPHNTTESRLVVDFSQ FSRGSTHVSWPKFAVPNLQSLTNLLSSNLSWLSLDVSAAFYHIPLHPA AMPHLLVGSSGLPRYVARLSSTSRNINYQHGTMQDLHDSCSRNLYVSL LLLYKTFGRKLHLYSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVR RAFPHCLAFSYMDDVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKT KRWGYSLNFMGYVIGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQ RIVGLLGFAAPFTQCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYL NLYPVARQRSGLCQVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAEL LAACFARSRSGAKLIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFV YVPSALNPAADPSAGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHL PDRVHFASPLHVAWRPPSKSPGSGPPMQLFHLCLIISCSCPTVQASKL CLGWLWGMDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYREA LESPEHCSPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSYV NVNMGLKIRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPNA PILSTLPETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQCMD AMKRGLCCVLLLCGAVFVSPSQEIHARFRRMGGWSSKPRQGMGTNLSV PNPLGFFPDHQLDPAFGANSNNPDWDFNPNKDHWPEANQVGASKGGKS GMENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPT CPGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLL VLLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPS DGNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVW LSVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI 7.3.MVA-SIi-HBV-PreS-P.sub.mut-C-TPA-S(sh): DescriptionforaminoacidsequencesofHBV immunogen 7.3.1.1.Firstaminoacidofpolypeptide=M 7.3.1.2.SharkInvariantchain(SIi)= (SEQIDNO:4) SLLWGGVTVLAAMLIAGQVASVVFLV 7.3.1.3.Linker= (SEQIDNO:31) SKSGPPSGKS 7.3.1.4.NPreS1andPreS2= (SEQIDNO:15) NSNNPDWDFNPNKDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGI LTTVPAAPPPASTNRQSGRQPTPISPPLRDSHPQAMQWNSTTFHQALL DPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSRTGDPAPN 7.3.1.5.Linker= (SEQIDNO:32) GSKSGSK 7.3.1.6.Pmut=(mutations:Y63A,C323A,C334A, C338A,C352A,R714A,D788A,R792A) (SEQIDNO:8) MPLSYQHFRKLLLLDDEAGPLEEELPRLADEGLNRRVAEDLNLGNLNV SIPWTHKVGNFTGLASSTVPVFNPEWQTPSFPHIHLQEDIINRCQQYV GPLTVNEKRRLKLIMPARFYPNLTKYLPLDKGIKPYYPEHAVNHYFKT RHYLHTLWKAGILYKRETTRSASFCGSPYSWEQELQHGRLVFQTSTRH GDESFCSQSSGILSRSPVGPCVRSQLKQSRLGLQPQQGSLARGKSGRS GSIRARVHPTTRRSFGVEPSGSGHIDNSASSTSSCLHQSAVRKTAYSH LSTSKRQSSSGHAVELHNIPPSSARSQSEGPIFSAWWLQFRNSKPASD YALTHIVNLLEDWGPATEHGEHNIRIPRTPARVTGGVFLVDKNPHNTT ESRLVVDFSQFSRGSTHVSWPKFAVPNLQSLTNLLSSNLSTFGRKLHL YSHPIILGFRKIPMGVGLSPFLLAQFTSAICSVVRRAFPHCLAFSYMD DVVLGAKSVQHLESLFTSITNFLLSLGIHLNPNKTKRWGYSLNFMGYV IGSWGTLPQEHIVLKIKQCFRKLPVNRPIDWKVCQRIVGLLGFAAPFT QCGYPALMPLYACIQSKQAFTFSPTYKAFLCKQYLNLYPVARQRSGLC QVFADATPTGWGLAIGHRAMRGTFVAPLPIHTAELLAACFARSRSGAK LIGTDNSVVLSRKYTSFPWLLGCAANWILRGTSFVYVPSALNPAADPS AGRLGLYRPLLHLPFRPTTGRTSLYAVSPSVPSHLPDRVHFASPLHVA WRPP 7.3.1.7.Linker= (SEQIDNO:33) SKSPGSGPP 7.3.1.8.C= 7.3.1.8.1.PreCore= (SEQIDNO:16) MQLFHLCLIISCSCPTVQASKLCLGWLWG 7.3.1.8.2.Core= (SEQIDNO:6) MDIDPYKEFGASVELLSFLPSDFFPSIRDLLDTASALYREALESPEHC SPHHTALRQAILCWGELMNLATWVGSNLEDPASRELVVSYVNVNMGLK IRQLLWFHISCLTFGRETVLEYLVSFGVWIRTPPAYRPPNAPILSTLP ETTVVRRRGRSPRRRTPSPRRRRSQSPRRRRSQSRESQC 7.3.1.9.TPA= (SEQIDNO:30) MDAMKRGLCCVLLLCGAVFVSPSQEIHARFRR 7.3.1.10.S(sh)_= 7.3.1.10.1.CPreS1= (SEQIDNO:21) MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVG 7.3.1.10.2.Linker= (SEQIDNO:34) ASKGGKSG 7.3.1.10.3.Surface(S)= (SEQIDNO:18) MENTTSGFLGPLLVLQAGFFLLTRILTIPQSLDSWWTSLNFLGGAPTC PGQNSQSPTSNHSPTSCPPICPGYRWMCLRRFIIFLFILLLCLIFLLV LLDYQGMLPVCPLLPGTSTTSTGPCKTCTIPAQGTSMFPSCCCTKPSD GNCTCIPIPSSWAFARFLWEWASVRFSWLSLLVPFVQWFVGLSPTVWL SVIWMMWYWGPSLYNILSPFLPLLPIFFCLWVYI 7.4.NucleotidesequencesofMVA-SIi-HBV-PreS- P.sub.mut-C-TPA-S(sh)isprovidedasSEQIDNO:27 7.5.Descriptionfornucleotidesequencesof MVA-SIi-HBV-PreS-P.sub.mut-C-TPA-S(sh): 7.5.1.F11-L-Flank=bases1-1097 (SEQIDNO:35) 7.5.2.SIi-HBV-PreS-P.sub.mut-C=bases1098-4838 (SEQIDNO:37) 7.5.3.Transcriptionterminatorsequence=bases 4839-4845 TTTTTGT 7.5.4.mH5promoter=bases4846-4942 (SEQIDNO:28) 7.5.5.TPA=bases4943-5038(SEQIDNO:29) 7.5.6.S(sh)=bases5039-5920(SEQIDNO:38) 7.5.7.F11-R-Flank=bases5921-7239 (SEQIDNO:36) 8.NucleotidesequencesoflowGCcontent versionMVA-SIi-HBV-PreS-Pmut-C-TPA-S(sh) (SEQIDNO:58): gtaatctattcgatataccgttgctaacagtatactggcccaataact gtggatggaaaatctataataatacattaatatcatccgatggtgcta gggttatttggatggatgcgtataaattttcttgcggtttatctttac aagactattgttatcattggggtagcaaaccagagagccgaccattcg atttaataaaaaaatcagatgctaaacgcaattctaaatcgttggtca aagaatctatggcatccttgaaatccttgtacgaggcattcgagacac aatcaggagcgttagaagttttaatgagtccatgtaggatgttttcgt tttctagaatagaagacatgttcttaactagtgtcattaatagagtat ccgagaatactggaatggggatgtattatcctaccaacgatatacctt ctctatttatcgaatcatctatctgtctagattatattatagtaaata atcaggaatccaacaaatatcgtatcaaatctgttctcgatatcattt cttcaaaacaataccctgcaggacgtcccaactacgttaaaaatggta caaaaggaaagttatatatcgcgttgtgtaaagttaccgtacctacta acgaccatattccagtagtttatcacgatgatgacaatactaccacct ttattacagtattgacgtccgtcgatattgaaactgctatcagagcag gatattcgatagtcgaattaggggctttacaatgggataataatattc cagaacttaaaaacggtttactggatagtatcaagatgatttatgact tgaacgcagttacaacaaataatttattggaacagctcatagaaaata ttaactttaacaactctagtataatttcgttgttttatacatttgcca ttagttattgccgagcattcatttactcaattatggaaaccatagatc cggtgtatatatctcagttcagttataaagaattatacgttagtagct cttataaagatattaatgaatccatgagtcagatggtaaaattataaa aagtgaaaaacaatattatttttatcgttggttgttacactATGTCAC TTCTTTGGGGCGGAGTTACAGTTCTTGCTGCTATGCTTATTGCTGGAC AAGTTGCTTCTGTTGTGTTTCTTGTTTCTAAATCTGGACCTCCTTCTG GAAAATCTAATTCTAACAATCCTGATTGGGATTTCAATCCTAACAAAG ATCATTGGCCTGAAGCTAATCAAGTTGGAGCTGGTGCTTTTGGACCTG GTTTTACACCTCCTCATGGTGGATTGCTTGGATGGTCACCTCAAGCTC AGGGAATTCTTACAACAGTTCCAGCTGCTCCTCCTCCTGCTTCTACAA ATAGACAATCTGGTAGACAACCTACACCTATTTCTCCACCTCTTAGAG ATTCTCATCCTCAAGCTATGCAATGGAATTCTACTACATTTCATCAAG CTTTGCTTGATCCTAGAGTTAGAGGACTTTATTTTCCTGCTGGTGGAA GTTCTTCTGGAACAGTTAATCCTGTTCCTACAACAGCTTCTCCAATTT CTTCTATATTTTCTAGAACAGGCGATCCTGCTCCTAATGGATCAAAAT CTGGATCAAAAATGCCTCTGTCTTATCAACACTTTAGAAAATTGCTGC TTCTTGATGATGAAGCTGGACCTCTTGAAGAAGAATTGCCTAGACTTG CTGATGAAGGACTTAATAGAAGAGTTGCTGAAGATCTTAATCTGGGAA ATCTTAATGTTTCTATTCCTTGGACACACAAAGTTGGAAATTTCACAG GACTTGCATCTTCTACAGTGCCTGTTTTTAATCCTGAATGGCAAACAC CTTCTTTTCCACATATTCATCTGCAAGAGGATATCATCAATAGATGTC AACAATATGTTGGACCACTGACAGTTAATGAGAAGAGAAGGCTTAAAC TTATTATGCCTGCTAGATTCTATCCTAATCTTACAAAGTATTTGCCTC TGGATAAGGGAATCAAACCTTATTATCCTGAACATGCTGTGAATCACT ACTTTAAAACAAGACATTATCTGCATACACTGTGGAAAGCTGGTATTC TTTACAAAAGAGAAACAACAAGATCTGCTTCATTTTGTGGATCTCCAT ATTCTTGGGAACAAGAACTTCAACATGGTAGACTTGTTTTTCAAACAT CTACAAGACATGGGGATGAATCATTTTGTTCTCAAAGTTCTGGAATTC TTTCTAGATCTCCTGTTGGACCTTGTGTTAGATCTCAACTTAAACAAT CTAGACTTGGACTTCAACCTCAACAAGGATCTCTTGCTAGAGGAAAAA GTGGAAGATCTGGATCTATTAGAGCTAGAGTTCATCCTACAACTAGAA GATCTTTTGGAGTTGAACCTTCTGGATCTGGACATATTGATAATTCTG CCTCTTCTACATCTTCTTGTCTGCATCAATCTGCTGTTAGAAAGACAG CTTATTCTCATTTGTCTACTTCTAAGAGACAATCATCTTCTGGACATG CTGTTGAACTTCATAATATTCCTCCAAGTAGTGCTAGAAGTCAATCTG AAGGACCAATATTTTCAGCTTGGTGGCTTCAATTCAGAAATTCTAAAC CTGCTTCTGATTATGCTCTGACACATATAGTTAATTTGCTTGAAGATT GGGGACCTGCTACAGAACATGGCGAACACAATATTAGAATACCTAGAA CTCCTGCTAGAGTTACAGGCGGAGTCTTTTTGGTTGATAAGAATCCTC ATAATACCACAGAATCAAGACTTGTTGTTGATTTTTCACAGTTTTCTA GAGGATCTACACATGTTTCTTGGCCTAAATTTGCTGTTCCAAATCTTC AATCTCTTACAAATTTGCTTTCATCTAATCTTTCTTGGCTGTCTCTTG ATGTTTCTGCTGCCTTTTATCATATTCCTCTTCATCCTGCTGCAATGC CTCATTTGCTTGTTGGATCATCTGGACTTCCAAGATATGTTGCTAGAC TTAGCTCTACATCTAGAAATATCAATTATCAGCATGGAACAATGCAGG ATCTTCACGATTCTTGTAGTAGGAATCTGTATGTTTCTTTGCTTCTGC TGTATAAGACATTTGGAAGAAAACTTCATCTGTATTCTCACCCTATTA TTCTGGGTTTTAGAAAGATTCCTATGGGAGTTGGACTTTCTCCTTTTT TGCTTGCTCAATTCACATCTGCTATTTGTTCTGTTGTTAGAAGGGCTT TTCCTCATTGTCTTGCATTTTCTTATATGGATGATGTTGTTCTTGGAG CTAAATCTGTTCAACATCTTGAAAGTCTGTTTACCTCTATTACTAATT TTCTGCTTTCTCTGGGAATTCATCTGAATCCAAACAAAACAAAGAGAT GGGGATATTCTCTTAATTTCATGGGATATGTTATTGGATCTTGGGGAA CACTTCCTCAAGAACATATCGTTTTGAAAATCAAGCAATGTTTCAGAA AACTGCCTGTGAATAGACCTATTGATTGGAAAGTTTGTCAAAGAATTG TGGGACTTCTTGGATTTGCTGCTCCTTTTACACAATGTGGATATCCTG CTCTTATGCCACTTTATGCTTGTATTCAATCTAAACAGGCTTTTACAT TTTCTCCAACATACAAAGCTTTTCTGTGTAAACAGTATCTGAATCTTT ATCCTGTGGCTAGACAAAGATCTGGTCTTTGTCAAGTTTTTGCTGATG CTACACCAACAGGATGGGGACTTGCTATTGGACATAGAGCTATGAGAG GAACATTTGTTGCTCCATTGCCTATTCATACAGCTGAATTGCTTGCTG CTTGTTTTGCTAGATCTAGAAGCGGAGCAAAACTTATTGGTACAGATA ATAGTGTTGTCCTGAGTAGAAAGTACACATCTTTTCCATGGTTGTTGG GATGTGCTGCTAATTGGATTCTTAGAGGAACTTCTTTTGTTTATGTTC CTTCTGCTCTTAATCCTGCAGCTGATCCATCTGCTGGTAGATTGGGAC TGTATAGACCACTTCTTCATTTGCCTTTTAGACCAACAACTGGAAGAA CATCTCTTTATGCTGTTTCTCCTTCTGTTCCATCTCATTTGCCTGATA GAGTTCATTTTGCTTCTCCACTTCATGTTGCTTGGAGGCCACCATCTA AATCTCCAGGTTCTGGACCACCTATGCAACTTTTTCATTTGTGTTTGA TCATTAGCTGTTCTTGTCCTACAGTTCAAGCTTCTAAACTTTGTCTTG GATGGCTTTGGGGAATGGATATTGATCCATACAAAGAATTTGGAGCTA GTGTTGAATTGCTGTCATTTCTTCCATCTGATTTTTTCCCTTCTATTC GTGATCTTCTTGATACAGCATCTGCTCTGTATAGAGAAGCTCTTGAAT CTCCTGAACACTGTTCTCCACATCATACAGCACTTAGACAAGCTATTC TTTGTTGGGGAGAACTTATGAATCTTGCTACATGGGTTGGATCTAATT TGGAAGATCCAGCTTCTAGAGAATTGGTGGTTTCTTATGTTAATGTGA ATATGGGACTGAAAATTAGACAACTGCTTTGGTTTCATATCTCTTGTC TTACATTTGGTAGAGAAACAGTTTTGGAATATTTGGTTTCTTTTGGCG TTTGGATTAGAACACCTCCAGCTTATAGACCTCCTAATGCTCCTATTT TGTCTACACTTCCTGAAACAACAGTCGTTAGAAGAAGAGGAAGATCTC CAAGAAGAAGAACACCAAGTCCTAGAAGAAGAAGATCTCAATCACCAA GAAGAAGAAGAAGTCAATCTAGAGAATCTCAATGTTGATTTTTGTTCG ACATTAAAAATTGAAAATAAATACAAAGGTTCTTGAGGGTTGTGTTAA ATTGAAAGCGAGAAATAATCATAAATATGATTCAGGTGACGGATCCAT GGATGCTATGAAGCGAGGACTTTGTTGTGTTTTGCTTCTTTGTGGTGC TGTGTTTGTTTCTCCATCTCAAGAAATTCATGCCAGATTCAGAAGAAT GGGAGGCTGGTCATCTAAACCTAGACAAGGCATGGGAACAAATCTTTC TGTTCCTAATCCTTTGGGATTCTTTCCTGATCATCAATTGGATCCAGC ATTTGGAGCAAATAGTAACAATCCAGATTGGGACTTTAACCCAAACAA AGATCATTGGCCAGAAGCTAATCAAGTTGGAGCATCTAAAGGTGGAAA AAGTGGAATGGAAAACACTACATCTGGATTTCTTGGACCTTTGCTTGT TCTTCAAGCTGGATTTTTCCTGTTGACAAGAATACTTACAATTCCTCA ATCACTGGATTCTTGGTGGACAAGTCTTAATTTTCTTGGAGGTGCTCC TACATGTCCTGGACAAAATTCTCAATCTCCAACTTCTAATCATTCTCC TACATCTTGTCCTCCAATTTGTCCTGGATATAGATGGATGTGTCTTAG AAGATTCATTATCTTTCTTTTCATACTGCTGCTGTGTCTGATTTTCCT TCTTGTTTTGTTGGATTATCAGGGAATGCTTCCTGTTTGTCCTTTGCT TCCTGGAACTTCTACAACAAGTACAGGACCTTGTAAAACATGTACAAT TCCAGCACAGGGAACATCTATGTTTCCAAGTTGTTGTTGTACAAAACC TTCTGATGGAAATTGCACATGTATTCCTATTCCAAGTTCTTGGGCATT TGCTAGATTTCTTTGGGAATGGGCTTCTGTTAGATTCAGTTGGTTGTC TCTTTTGGTTCCATTTGTTCAGTGGTTTGTTGGATTGTCTCCTACAGT TTGGCTTTCTGTTATTTGGATGATGTGGTATTGGGGACCTTCTCTTTA CAATATTTTGAGTCCTTTTCTCCCTTTGCTGCCAATTTTCTTTTGTCT TTGGGTTTACATTTGAttaaccgagtttctgcattattgtaattcgta tgctggcaccatcaaagaatcacttctaaaagatatcaatatcacaca tacaaatattactaccctattgaatgagacagccaaggttatcaagtt agtaaaatctctggtagataaagaagatactgatattgtgaataattt cattaccaaagaaattaaaaacagagacaaaatagttaatagtttgtc tctatcaaacctggactttcgtttgtaaattggggctTtttgtacaat aaatgggtgttgccaatgattcatcccctgaatatcaatggatgtctc cccatagattatcagatactgttatattaggagactgtttgtatttta acaatataatgtcccaattagatttacaccaaaattgggctccatcag ttagattgttaaattattttaagaattttaataaggaaacactactaa agatagaagagaatgattacattaattcatcctttttccaacaaaagg ataaacgattttatcctataaacgacgatttttatcacatatctacag gaggatatggtatagtctttaagatagataactatgtagtaaaatttg tattcgaggccacaaaattatatagtcccatggaaactacggcggagt tcacagtacccaaatttctatacaacaatctaaagggagatgaaaaaa aattaatcgtgtgtgcgtgggccatgggattaaactataaattaacat ttttacatactctgtataaacgtgttcttcatatgttgctattattga tacaaactatggatggtcaggaactatcattgagatattcttctaaag tttttttaaaggcgtttaacgagagaaaggacagtatcaaattcgtga aattactatcccacttttatccggcagttattaacagtaatattaatg ttataaactattttaaccgcatgtttcactttttcgaacatgaaaaga gaactaactacgaatacgaaagaggaaatattataatttttcccctag cactgtattcggcagataaagtagataccgagctagctatcaaattag gatttaaatctttggtacaatacataaagtttatctttttacagatgg ctctgttatacattaaaatttacgaactaccatgctgcgacaactttt tacacgcagatcttaaacccgataatatcttactttttgattccaatg aaccaataataattcatctaaaggataaaaagtttgtttttaatgaac gtattaaatcggcattaaacgactttgacttttcccaag

    [0217] 8.1 Description for nucleotide sequences of low GC content version of MVA-SIi-HBV-PreS-Pmut-C-S(sh): [0218] 8.1.1 F11-L-Flank=bases 1-1097 (SEQ ID NO: 35) [0219] 8.1.2 SIi-HBV-PreS-Pmut-C=bases 1098-4838 (SEQ ID NO: 59) [0220] 8.1.3 Transcription terminator sequence=bases 4839-4845 TTTTTGT [0221] 8.1.4 mH5 promoter=bases 4846-4942 (SEQ ID NO: 28) [0222] 8.1.5 TPA=bases 4943-5038 (SEQ ID NO: 60) [0223] 8.1.6 S(sh)=bases 5039-5920 (SEQ ID NO: 61) [0224] 8.1.7 F11-R-Flank=bases 5921-7239 (SEQ ID NO: 36)

    [0225] The Sequence of wild-type HBV Polymerase is provided as SEQ ID NO: 19.

    [0226] ChAdOx1 Sequence

    [0227] ChAdOx1 sequence 5 to the immunogen cassette is provided as SEQ ID NO: 39.

    [0228] ChAdOx1 sequence 3 to the immunogen cassette is provided as SEQ ID NO: 40.

    [0229] ChAdOx2 Sequence

    [0230] ChAdOx2 sequence 5 to the immunogen cassette is provided as SEQ ID NO: 41.

    [0231] ChAdOx2 sequence 3 to the immunogen cassette is provided as SEQ ID NO: 42.

    [0232] MVA Sequence

    [0233] MVA sequence 5 to the immunogen cassette is provided as SEQ ID NO: 44. MVA sequence 3 to the immunogen cassette is provided as SEQ ID NO: 45

    [0234] The CMV long promoter with Tetron Operator sequence is provided as SEQ ID NO: 50. The CMV short promoter with Tetron Operator sequence is provided as SEQ ID NO: 51.

    TABLE-US-00008 PreS1sequence(SEQIDNO:52): MGGWSSKPRQGMGTNLSVPNPLGFFPDHQLDPAFGANSNNPDWDFNPN KDHWPEANQVGAGAFGPGFTPPHGGLLGWSPQAQGILTTVPAAPPPAS TNRQSGRQPTPISPPLRDSHPQA PreS2sequence(SEQIDNO:53): MQWNSTTFHQALLDPRVRGLYFPAGGSSSGTVNPVPTTASPISSIFSR TGDPAPN