Herpes Simplex Virus Type 1 Infected Cells Research Articles

Adeno-associated virus DNA replication complexes in herpes simplex virus or adenovirus-infected cells.

A complex which is active in in vitro synthesis of adeno-associated virus (AAV) DNA was solubilized from Vero cells that were co-infected with AAV and either adenovirus (Ad5) or a herpes simplex virus type 1 (HSV-1) as the helper virus. The complexes from the Ad5 and HSV-1-infected cells sedimented at 23 S and 28 S, respectively. The optimal conditions for in vitro DNA synthesis for the two types of complex using the endogenous AAV template and the endogenous DNA polymerase, differed with respect to the effect of KCl and K2SO4 concentration. In addition the complex from HSV-1-infected cells, but not that from Ad5-infected cells, was inhibited by phosphonoacetic acid. Thus, the two complexes appear to contain different DNA polymerase activities. This was verified by phosphocellulose chromatography of the DNA polymerases solubilized from the isolated complexes. The major activity in the complex from HSV-1 infected cells was the HSV-induced DNA polymerase with lesser amounts of cellular DNA polymerase alpha and gamma or both. The complex from the Ad5-infected cells contained mainly a cellular DNA polymerase gamma.

Crossed immunoelectrophoresis of a herpes simplex virus type 1-specific antigen: immunological and biochemical characterization.

Crossed immunoelectrophoresis of herpes simplex virus (HSV) antigens obtained from infected cells solubilized with the nonionic detergent Triton X-100 has shown that antigen number 6 (Ag-6) is found in preparations from HSV type 1-infected cells but not in HSV type 2 preparations. A monospecific antiserum was produced against Ag-6 by intracutaneous inoculation of the corresponding immunoprecipitate into rabbits. This antiserum neutralized HSV type 1 but not HSV type 2. The polypeptide composition of Ag-6 (molecular weight, 131,000 daltons) was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the immunoprecipitate, using radiolabeled antigen. The multitude of information obtained demonstrates that crossed immunoelectrophoresis can be used preparatively, since specific immunoprecipitates represent a short cut for the attainment of immunoreactive viral protein entities useful for further immunological and biochemical analyses.

Complement‐dependent cytotoxic antibodies in the course of cervical carcinoma

Complement-dependent cytotoxic antibodies to three cervical carcinoma cell lines (Me-180, SW-732 and HeLa) and to Herpes simplex virus type 2 (HSV-2)-infected cells were determined in a long-term study of women with cervical carcinoma. Cytolysis of surface antigens differed significantly between the cervical carcinoma cell lines and HSV-2. Regression of the tumor during treatment was accompanied by decreasing cytolysis in the cervical carcinoma lines, while tumor bearers and patients who became severely ill had high or increasing cytotoxic antibodies, The opposite was noted for the antibody response to HSV-2 infected cells: patients with less advanced cancer had significantly higher cytolytic activity than those who were severely ill or had advanced cancer; long-term survivors demonstrated high, stable lysis of HSV-2 infected cells. As a control of the cervical carcinoma cell lines, cytolytic activity was tested also on a lung carcinoma cell line, A-549. No significant differences were found between the patients with cervical cancer and the control women. Tumor bearers and patients treated for advanced cervical cancer showed a slight but non-significant increase in cytolytic activity on the A-549 line.

DNA Polymerase in Pseudorabies Virus Infected Cells

The DNA polymerase activity in BHK 21/C13 cells infected with pseudorabies virus is inhibited by incubation with antiserum to pseudorabies but not by incubation with pre-immune serum or by antiserum to herpes simplex virus type 1 (HSV-1) or herpes simplex virus type 2 (HSV-2). It also differs from the cell enzyme and the enzymes in HSV-1 or HSV-2 infected cells in its requirement for KCl in the in vitro assay. It seems likely, therefore, that pseudorabies virus specifies its own DNA polymerase.