Characterization of glycoprotein C of HSZP strain of herpes simplex virus 1

Sequences of UL44 genes of strains HSZP, KOS and 17 of herpes simplex virus 1 (HSV-1) were determined and the amino acid sequences of corresponding glycoproteins (gC) were deduced. In comparison with the 17 strain, the HSZP strain showed specific changes in 3 nucleotides and in 2 amino acids (aa 139...

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Published in:	Acta virologica (Anglickâa verze) Vol. 44; no. 2; p. 99
Main Authors:	Oravcová, I, Kúdelová, M, Mlcuchová, J, Matis, J, Bystrická, M, Westra, D F, Welling-Wester, S, Rajcáni, J
Format:	Journal Article
Language:	English
Published:	Slovakia 01-04-2000
Subjects:	Animals Antibodies, Monoclonal - immunology Antibodies, Viral - immunology Baculoviridae Base Sequence Cell Line Cercopithecus aethiops DNA, Viral Enzyme-Linked Immunosorbent Assay - methods Gene Expression Genetic Vectors Herpesvirus 1, Human - chemistry Herpesvirus 1, Human - immunology Herpesvirus 1, Human - isolation & purification Humans Molecular Sequence Data Spodoptera Vero Cells Viral Envelope Proteins - chemistry Viral Envelope Proteins - genetics Viral Envelope Proteins - immunology
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Summary:	Sequences of UL44 genes of strains HSZP, KOS and 17 of herpes simplex virus 1 (HSV-1) were determined and the amino acid sequences of corresponding glycoproteins (gC) were deduced. In comparison with the 17 strain, the HSZP strain showed specific changes in 3 nucleotides and in 2 amino acids (aa 139 and 147, both from Arg to Trp) in the antigenic locus LII. The change at aa 147 was situated within the GAG-binding epitope. In a similar comparison, KOS strain had changes in 3 nucleotides and 3 amino acids (aa 3, 14, and 300). The UL44 genes of HSZP and KOS strains were expressed in insect Sf-21 cells by means of the baculovirus (Bac-to-Bac) expression system. As shown by immunoblot analysis, both the recombinant baculoviruses (B1-HSZP and B6-KOS) expressed a glycosylated gC, the M(r) of which (116 K) was lower than that of gC synthesized in Vero cells (129 K) infected with strains HSZP or KOS. In addition, smaller gC-specific proteins (of apparent M(r) of 50-58 K and 98 K) corresponding to a non-glycosylated precursor polypeptide and/or incomplete forms of the partially glycosylated gC were found. When Balb/c mice were immunized with Sf-21 cells expressing gC, the recombinant gC-HSZP represented a more efficient immunogen possibly due to its stronger expression in these cells. The corresponding gC-HSZP antiserum reacted in enzyme-linked immunosorbent assay (ELISA) equally well with HSZP and KOS virion antigens and neutralized HSZP strain at a low titer. Both gC-HSZP and gC-KOS antisera detected the homologous as well as the heterologous gC antigens in Vero cells regardless whether infected with strains HSZP, KOS or 17, revealing the presence of gC from 6 to 16 hrs post infection (p.i.) in the cytoplasm, on the nuclear membrane and at the cell surface.
ISSN:	0001-723X