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Immunity to Virus-Free Syngeneic Tumor Cell Transplantation in the BALB/c Mouse after Immunization with Homologous Tumor Cells Infected with Type C Virus¹

From the Microbiological Associates, Walkersville, Maryland 21793; Department of Microbiology, School of Medicine, George Washington University, Washington, D.C. 20006; and the Viral Immunology Section, Laboratory of RNA Tumor Viruses, National Cancer Institute, National Institutes of Health, Public Health Service, United States Department of Health, Education, and Welfare, Bethesda, Maryland 20014

Abstract

Syngeneic tumor cell lines free of endogenous type C virus or viral antigen expression were derived from spontaneously occurring tumors of the BALB/cCr mouse. Two cell lines free of endogenous type C virus were examined and found to be highly tumorigenic in tumor growth kinetic studies. In vitro inoculation of these cell lines with Rauscher-murine leukemia virus (R-MuLV) resulted in their chronic infection in which 95 to 100% of the cells were scored as virus positive. These infected lines showed a highly significant increase in their immunogenicity as compared to their uninfected controls. Animals in which these virus-positive tumors regressed were then shown to be highly resistant to challenge with the uninfected tumor cell lines as well as to live R-MuLV. This observed resistance to uninfected tumor cell lines could not be induced by immunization of the mouse with uninfected tumor cells and R-MuLV simultaneously at the same injection site, nor could it be induced with lethally irradiated virus-infected tumor cells, subtumorigenic doses of uninfected cells, or inactivated R-MuLV or Gross leukemia virus (G-MuLV). Cell-mediated cytotoxicity studies revealed that spleen cells obtained from animals whose virus-infected tumors regressed were cytotoxic to homologous infected and uninfected tumor cells as well as to other uninfected tumor cell lines syngeneic to the BALB/c mouse. Correlation of in vitro cytotoxicity with in vivo immunity was provided by the Winn assay, by inoculation into susceptible mice of immune and nonimmune spleen cells premixed with uninfected tumor cells. The immune cells were highly effective in preventing this tumor cell transplantation. It was concluded that type-C virus infection of these syngeneic tumor cells resulted in their acquiring strong transplantation antigens that were in part due to the virion, but were at least in part due to alterations of antigens or haptens that are present in a less immunogenic form on the uninfected tumor cell.

Footnotes

¹ Supported by Public Health Service Contract N01-CP-33248 from the Division of Cancer Cause and Prevention within the Virus Cancer Program of the National Cancer Institute.

² This paper was part of a dissertation submitted to the Graduate School of Arts and Sciences, The George Washington University, in partial fulfillment of the requirement for the Doctor of Philosophy degree.

³ Present address: Microbiological Associates, Building 1, Walkersville, Maryland 21793.