A simple method for immune analysis of proteins following sodium dodecyl sulphate polyacrylamide gel electrophoresis.

Immune analysis of polypeptides separated by SDS-PAGE provides a technique useful in the characterisation of complex mixtures. We describe a simple procedure involving immune analysis of fractionated PAGE gels. Polypeptides of purified herpesvirus simplex, HSV 'excreted antigen' and cell culture 'control' preparations were separated by SDS-PAGE. Following electrophoresis, protein was eluted from gels, adsorbed to plastic and analysed using an RIA technique. When using a mouse anti-HSV1 serum, 7 peaks of immune reactivity were observed with pure virus and 6 with HSV 'excreted antigen'. Only 1 peak of reactivity was observed with a 'control' antigen.

Production of herpes simplex virus fluorescein labelled typing reagents.

29 herpes simplex virus monoclonal antibodies (MAbs) were produced and shown by indirect immunofluorescence assay (IIFA) to be HSV type specific or HSV cross reactive. Two HSV-1 and two HSV-2 specific MAbs were selected as HSV typing reagents on the basis of their specificity and the fluorescence pattern produced. When directly conjugated to fluorescein both reagents detected and correctly identified HSV isolates previously typed by restriction endonuclease analysis (REA). No cross reactions were observed with a panel of human viral pathogens. Assessment of these reagents in parallel with the Syva Microtrak Culture Identification/Typing Test indicated 100% concordance for the detection and typing of 80 clinical isolates.

Herpesvirus simiae (B virus) antibody response and virus shedding in experimental primary infection of cynomolgus monkeys.

Four cynomolgus monkeys (Macaca fascicularis) were inoculated in the lips and tongues with B virus. Virus shedding and antibody responses were monitored for up to 50 days postinfection. Virus was isolated from the oral cavities of all monkeys at 6 days postinfection despite the absence of observable lesions. Virus was not isolated from genital swabs or serum. Antibodies to both B virus and herpes simplex virus were detected by neutralization between days 8 and 12. Virus-specific IgM and IgG antibodies were measured by antibody capture radioimmunoassay. IgM was first detected on day 6; by contrast, IgG did not appear until day 12. Antibodies reactive in a competitive radioimmunoassay appeared by day 12 and peaked at 30 to 40 days postinfection. This study provides data on which to base the diagnosis of primary B virus infection in cynomolgus monkeys.

Monoclonal antibodies for the identification of herpesvirus simiae (B virus).

To differentiate between B virus and HSV isolates from monkeys and man monoclonal antibodies (mabs) were produced to herpesvirus simiae (B virus) and herpes simplex type 1 and 2 (HSV-1 and HSV-2). Mabs were tested by indirect immunofluorescence (IFAT) for reactivity against herpesviruses from Asiatic monkeys (B virus), African monkeys (SA 8 virus), and man (HSV-1, HSV-2, varicella-zoster virus, cytomegalovirus, and Epstein-Barr virus). Mabs could be divided into groups A-E displaying specific reactivity for B virus (A); reactivity with both B virus and SA 8 but not HSV (B); reactivity with B virus, SA 8 virus and HSV strains (C); specific reactivity with HSV-1 (D); and specific reactivity with HSV-2 (E). Two of the B virus specific mabs were able to differentiate between cynomolgus and rhesus strains of B virus. None of the mabs reacted with human varicella-zoster virus, cytomegalovirus, or Epstein-Barr virus. A panel of mabs for the unequivocal identification of B virus isolates from monkey or man is proposed.