Inducible expression of herpes simplex virus type 1 glycoprotein C in NIH 3T3 cells

Antibodies directed against the glycoprotein C (gC) of herpes simplex virus type 1 (HSV-1) are known to be mainly HSV-1-specific, whereas antibodies against other major HSV-1 glycoproteins cross-react with HSV-2 antigens. To investigate the immunological features of gC-1, the gene encoding gC-1 was isolated and cloned from DNA of cells infected with HSV-1. The 3.6 kbp SalI fragment R of HSV-1 DNA was modified in order to place the gene under transcriptional control of the glucocorticoid dependent promoter of the MMTV-LTR. NIH 3T3 cells were transfected with the resulting plasmid. Cell lines established by selection for the vector-encoded marker gene were tested for the ability to synthesize gC-1 after addition of dexamethasone to the growth medium. Glycoprotein-enriched cell extracts of several clones were shown to contain gC-1 by immunoblotting using a gC-1-specific monoclonal antibody. One cell line was used to show the presence of gC-1 also in the culture supernatant. gC-1 synthesis decreased after several passages of the cells but could be restimulated by the addition of 5-azacytidine to the cultures.