Preparation of immune serum to immediate early and early polypeptides specified by herpes simplex virus type 1.

An antiserum to the immediate early (IE) ana early (E) polypeptides specified in SIRC cells within 4 hr post infection (p.i.) with herpes simplex virus type 1 (HSV1) was prepared in rabbits by sequential injections of two different antigens. The first antigen, which contained the alpha polypeptides, was prepared from infected cells treated for 5 hr with cycloheximide and incubated in the presence of actinomycin D. The second antigen contained all polypeptides, synthesized for 4 hr after removal of cycloheximide. The 14C amino acid-labelled antigen extracts, pulsed for different intervals p.i., were precipitated with the serum to IE and E antigens, with a serum to purified viral particles and control (preimmune) serum. The antigen extracts and their precipitates were analyzed by polyacrylamide gel electrophoresis. The serum to IE and E antigens reacted at least with 3 alpha polypeptides (ICP 4, 0, 11) and several beta polypeptides (ICP 6, 8, 18, 26, 36 and 39). It also reacted with the capsid polypeptide ICP 5 (155,000) which was synthesized from 3 hr p.i. Anticomplementary immunofluorescence revealed bright granular staining in nuclei of VERO and SIRC cells between 3-6 hr p.i.