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Mechanism of Mastocytoma-Mediated Suppression of Lymphocyte Reactivity¹

From the Department of Microbiology, State University of New York, Upstate Medical Center, Syracuse, New York 13210

Abstract

Previously we have shown that mitomycin-treated P-815 (H-2^d) cells (P-815_m) inhibit the in vitro response of C57BL/6 spleen cells to mitogens and DBA/2 alloantigens. The present data indicate that P-815_m cells also inhibit the response of C57BL/6 spleen cells to AKR (H-2^k)-stimulating cells and that the inhibition does not appear to be the result of crowding. Subpopulations of spleen cells obtained by density gradient centrifugation or after removal of glass-adherent cells are all sensitive to the inhibitory effects of P-815_m. We also show that P-815_m can inhibit the in vivo generation of graft-versus-host reactivity. Thus the inhibition due to P-815_m cells appears to operate both in vitro and in vivo.

The experiments characterize the suppression of MLC reactivity by cell-free preparations of sonicated P-815 mastocytoma cells. The results suggest that the inhibition is not removed by ultracentrifugation, ultra-violet irradiation, or dialysis of the sonicate. P-815 sonicates subjected to chloroform extraction, heating at 56°C, 0.1 µ filtration, or treatment with anti-minute virus of mice serum are still inhibitory. This inhibition is resistant to RNAase treatment but destroyed by pronase. P-815 sonicate suppresses the response of C57BL/6 spleen cells to both DBA/2 and AKR-stimulating spleen. Inhibition of MLC reactivity is only seen if P-815 sonicate is added within the first 48 hr after the initiation of culture. These results indicate that the inhibition of MLC by sonicates of P-815 cells is due to a nondialyzable protein whose effects are not H-2 restricted and are acting on the early phase of sensitization.

Footnotes

¹ This investigation was supported by United States Public Health Service Research Grant CA 15462 from the National Cancer Institute.

² Part of these studies were submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy.

³ Send all correspondence to: Dr. Bertie F. Argyris, Department of Microbiology, State University of New York, Upstate Medical Center, Syracuse, New York 13210.