25 research outputs found
Analysis of glycoproteins expressed by isolates of herpes simplex virus causing different forms of keratitis in man
An in vitro analysis of glycoprotein produced by nine human ocular isolates of HSV-1 is reported. The source of the isolates was; three patients with recurrent dendritic keratitis, three with chronic stromal disease and three with primary keratoconjunctivitis. Virus strains were labelled with the radioactive precursors (35s) methionine and (14c) glucosamine. Radiolabelled viral glycoproteins were subsequently analysed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), followed by autoradiography. Viral glycoproteins were further characterised by immuno-precipitation with polyclonal and monoclonal antibodies to HSV. The stromal isolates excrete larger amounts of 'soluble' precursor glycoprotein D than those in the other two disease categories. It is possible that the immune response to glycoprotein D is in part responsible for the severity of stromal disease.</p
Herpesvirus simplex in chronic human stromal keratitis
A method is described for the isolation of Herpesvirus simplex (HSV) from corneal discs of patients suffering from chronic stromal keratitis. The discs were removed during penetrating keratoplasty. Virus was successfully isolated from 2 out of 8 discs maintained in vitro.</p
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.</p