The Journal of Medical Microbiology, Vol 25, Issue 1 49-57, Copyright © 1988 by Society for General Microbiology
The polyspecific immunoglobulin response to HSV-1 viral proteins: determination of immunogenic proteins and relative antibody titres to individual polypeptides by immunoblotting
R. R. McKendall, M. Pettit and W. Woo
Department of Neurology, University of Texas Medical Branch, Galveston 77550.
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.
