Hepatitis B virus proteins expressed by recombinant vaccinia viruses: influence of preS2 sequence on expression surface and nucleocapsid proteins in human diploid cells

Abstract

Fifteen vaccinia virus (VV) recombinants derived from VV strains Praha, LIVP and DD (i.e. Dryvax Wyeth vaccine-derived) and expressing genes for S, preS2-S or c antigens of hepatitis B virus (HBV) were tested in monkey CV-1 cells and human diploid LEP cells. The production of infectious virus was found to be alike in all the recombinants and parental viruses as well. However, several recombinants produced markedly lesser amounts of S and preS2 antigens in LEP cells than in CV-1 cells. This reduction was independent of the parental virus used. There was, however, a relationship between the production of preS2 in CV-1 cells and the production of S and preS2 antigens in LEP cells; in general, recombinants efficiently inducing preS2 antigen formation in CV-1 cells produced markedly reduced amounts of S and preS2 antigens in LEP cells. Reduction of HBV antigen production in LEP cells was not apparent in recombinants expressing only S or c antigens of HBV, and the production of c antigen by double recombinants was not influenced by simultaneous expression of preS2 and S. The various recombinants also differed in the ratio of S:preS2 antigen formation. This difference seemed to be associated with the length of the untranslated leader sequence preceding preS2 but not with the parental virus or cell type used. The titers of antibodies against S and preS2 antigens induced in mice immunized with different recombinants differed markedly. The differences in the ratio of S:preS2 antigen production in vitro were not reflected in vivo by S:preS2 antibody ratio.

Effect of virulence on immunogenicity of single and double vaccinia virus recombinants expressing differently immunogenic antigens: antibody-response inhibition induced by immunization with a mixture of recombinants differing in virulence

“…To prepare a plasmid for Table 1. Recombinants used and some of their properties S and gE antigens were quantified by ELISA as described previously (Kutinova! et al, 1994 ;Ludvı!…”

“… S and gE antigens were quantified by ELISA as described previously (Kutinová et al , 1994 ; Ludvíkov á et al , 1991 ); the absorbances for gE antigen from viruses 13 or 20 were subtracted as background from the absorbances of their recombinants. …”

Antitumor activity and immunogenicity of recombinant vaccinia virus expressing HPV 16 E7 protein SigE7LAMP is enhanced by high-level coexpression of IGFBP-3

“…The virus P13-preS2S with expression of hepatitis B virus envelope antigen has been described previously. 52 Viruses were propagated in BSC40 cells in Dulbecco's modified Eagle's medium (PAA, Pasching, Austria) supplemented with 10% fetal bovine serum (PAA). Virus (IMV) was purified by sucrose gradient centrifugation.…”

“…All three recombinant viruses induced cytotoxic T lymphocyte responses against the HPV16 E7 nonapeptide. [49][50][51][52][53][54][55][56][57] Coexpression of IGFBP-3 enhanced the E7-specific response only subtly. The enhancement of the T-cell response by IGFBP-3 coexpression was more apparent against the VACV E3-derived peptide in mice receiving P13-SigE7Lamp-H5-IGFBP-3 and P13-SigE7Lamp-E/L-IGFBP-3.…”

The expression of the soluble isoform of hFlt3 ligand by recombinant vaccinia virus enhances immunogenicity of the vector

“…The control virus P13-SigE7LAMP TK-was prepared by recombination with plasmid p11K which contains the TK gene disrupted by the 11k promoter and the multiple cloning site. The virus P13-pS2S with expression of hepatitis B virus envelope antigen was described previously (16). Recombinant P13 viruses were grown in BSC40 cells.…”