Theses and Dissertations

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Defense Date

6-1986

Document Type

Dissertation - NSU Access Only

Department

Oceanographic Center

First Advisor

Peter Andreotti

Second Advisor

Claire Ann Thuning

Third Advisor

Josephine Hurst

Fourth Advisor

Joel Warren

Abstract

The objective of these studies was to produce monoclonal antibodies against P388Dl cells, a lymphocytic leukemia of murine origin, to evaluate the efficacy of immunotherapy in tumor bearing DBA/2 mice.

Hybridomas producing anti-P388Dl MAbs were obtained by fusion between Sp2/0 myeloma and irrununized BALB/c spleen cells. Monoclonal antibodies were screened by enzyme linked immunoabsorbent and radioimmunoassays, and positive hybridomas were cloned and further subcloned by limiting dilution in fluid phase.

Four subclones designated G10, C9, C7, and D11 showed significant binding to both solubilized and live P388Dl cells but not to U937, Ehrlich, Ependymoblastoma, or L1210 tumor cells, or normal DBA/2 kidney, liver, heart, spleen, and thymus cells.

In complement dependent cytotoxicity assays, lysis of the P388Dl tumor cells was not observed. However, significant killing of P388Dl target cells was observed in antibody dependent cell-mediated cytotoxicity assays using DBA/2 spleen effector cells.

Two parameters, survival time and tumor volume were used to determine the efficacy of anti-P388Dl MAbs in irrununotherapy of tumor bearing DBA/2 mice. A significant prolongation of survival time and reduction of tumor size compared to untreated controls were observed in animals treated with MAbs G10 or C7. In mice treated with MAb C9 or a mixture of the antiP388Dl MAbs, a significant increase in survival time but no decrease in tumor size was observed. Mice injected with P388D1 tumor cells pretreated in vitro with the MAb mixture or MAb C7 survived longer than the untreated animals with no decrease in tumor size.

This study indicates the possibility of using MAbs G10, C7, and P388Dl leukemia cells as a model system for studying the effi6acy of MAb immunotherapy and the mechanism(s) involved in tumor cell destruction in vivo.

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