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Requirements for Induction of Specific Suppressor T Cells and Detection of their H‐2 Antigen‐binding Receptors by Fractionation on Target Cell Monolayers

The MC‐resistant specific suppressor T cells that inhibit DNA synthesis and CTL generation in MLC were induced in vitro by γ‐irradiated allogeneic lymphoid cells in a large dose. MLRs were inhibited only slightly when triggered by third‐party cells, even neighbouring with the corresponding sponding stimulators. Unlike the irradiated cells, intact allogeneic lymphoid cells induced a mixture of macrophage‐like and T‐cell suppressors with a pronounced non‐specific component of the action. Syngeneic cells induced low active non‐specific suppressors of macrophage type only. The suppression was not due to a cytotoxic effect, since specific T suppressors differed from CTL by conditions of induction and high sensitivity to γ‐irradiation and from CTL procurers by high sensitivity to CY and HC. The specific T‐suppressors could be selectively removed by adherence to a macrophage monolayer of the corresponding donor. The subsequent elution of adherent lymphocytes with pronase resulted in enrichment of specific T suppressors by a factor of 30 and 2.6, as judged by reduction in the number of lymphocytes required for 50% inhibition of DNA synthesis and 33% inhibition of CTL generation, respectively. The high specificity of this enrichment is shown by using both syngeneic monolayer for fractionation and third‐party stimulators in MLR for testing and by disappearance of slight non‐specific suppression caused by non‐fractionated suppresses. Complete inactivation of the eluted suppressors with anti‐Thy‐1.2 and anti‐T antibodies, their resistance to anti‐Mls antibodies, carrageenin, and carbonyl iron, together with the data of autoradiographic study and total DNA synthesis in the population indicate that the eluted highly specific suppressors represent mainly DNA‐synthesizing large and medium T lymphocytes.