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Splenic Suppressing Factor: Purification and Characterization of a Factor Suppressing Thymidine Incorporation into Activated Lymphocytes¹

From the Department of Medical Biology and Parasitology, Jichi Medical School, Tochigi-ken 329-04, Japan

Abstract

Suppressing activity upon the mitogen-activated lymphocytes was found in the supernatant (SUP) from the culture of mouse spleen, high-density subpopulation of thymocytes, and peritoneal exudate cells. Suppressing factor was obtained from the non-stimulated lymphocytes cultured for 24 to 36 hr with or without serum. Suppressing activity in the SUP was observed in the incorporation of ³H-thymidine, ³H-uridine, and ³H-leucine into Con A-activated lymphocytes or in the proliferation of L cells. Suppressing factor partially purified by Sephadex G-25 column chromatography was a heat-stable and dialyzable substance(s). Further purification and isolation of this factor by two-dimensional thin layer chromatography revealed that this was thymidine and thymidine monophosphate. The suppression in ³H-thymidine incorporation was attributed to the dilution effect of cold thymidine released from cultured lymphocytes.

Footnotes

¹ This investigation was supported in part by Research Grants 957051 and 911203 from the Ministry of Education of Japan.