The polyspecific immunoglobulin response to HSV-1 viral proteins: determination of immunogenic proteins and relative antibody titres to individual polypeptides by immunoblotting

We developed an immunoblotting procedure to characterise the polyspecific immunoglobulin response to the proteins of herpes simplex virus (HSV)-1. We found that 8-20% gradient polyacrylamide gels provided no advantage over fixed 8.5% gels for preparing Western blots for use in immunoblotting. The amount of protein loaded on the gels markedly influenced which proteins were detected by immune serum. The presence of Triton-X-100 0.5% in washes and buffers improved band clarity on immunoblots. In optimal conditions, immune mouse serum reacted with up to 33 HSV-1 lysate proteins. Six bands or regions appeared to be of major immunogenic reactivity, including the (122-130) x 10(3)-mol. wt region, a 75 x 10(3)-mol. wt protein, the gD region of approximately (56-64) x 10(3)-mol. wt and two non-glycosylated bands at mol. wts(10(3)) 42 and 44. Minor proteins, more weakly reactive, were detected at 27 other areas. The relative antibody titres in immune mouse serum to the different major regions showed antibody titre to gD greater than gB/gC greater than (42/44) x 10(3) much greater than P75 much greater than VP154. Most human sera reacted with all of the major and many of the minor immunogenic proteins but individual sera varied markedly in the proteins recognised. We conclude that immunoblotting is valuable for evaluating immunoglobulin responses to major and minor immunogenic proteins of HSV-1.