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.