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Influence of Antisera from Histoincompatible Mice on Electrophoretic Mobility of EL4 Tumor Cells¹

Department of Biochemistry, City of Hope Medical Center, Duarte, California 91010

Ascitic fluid containing antibodies to normal tissue cells and neoplastic cells (EL4 lymphoma) of C57BL/10J (H-2^b) mice was produced in a coisogenic B10.D2 (H-2^d) strain of mice by intraperitoneal injection of the tissue preparations together with adjuvant. -Globulins were precipitated by ammonium sulfate and subfractions separated on Sephadex G-200. These were examined by microelectrophoresis for effects on surface charges of EL4 lymphoma cells and for hemolytic potency in tests with histoincompatible erythrocytes. The neoplastic cells elicited production of relatively large quantities of a tumor-specific antibody, which in the presence of active complement reduced the electrophoretic mobility of EL4 tumor cells, but which was inactive in the hemolytic test. This antibody could not be removed by repeated treatment with normal tissue cells, but was absorbed specifically by the tumor cells. In the mice given neoplastic cells there was produced a relatively small quantity of (presumably) H-2 isoantibodies of which one subfraction did not influence the mobility of tumor cells but showed hemolytic activity, and of which another subfraction was active in both tests. The latter subfraction lost its mobility-reducing effect when heated to 58°C for 10 min. However, injection of normal tissues resulted in production of isoantibodies which were detectable only in the hemolytic test. A preliminary study was made of the different immunoglobulins in the analytic ultracentrifuge.

¹ This investigation was supported by Grant C-2568 from the National Cancer Institute, NIH.

² Present address: Aichi Cancer Center, Research Institute, Chikusa-Ku, Nagoya, Japan.

Received 8/15/66. Accepted 1/11/67.