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Proliferation of Stimulated Human Peripheral Blood Lymphocytes: Preferential Incorporation of Concanavalin a by Stimulated Cells and Mitogenic Activity¹

From the Department of Microbiology, The University of Chicago, Chicago, Illinois 60637

Abstract

Binding of concanavalin A (conA) to human peripheral blood lymphocytes or to lymphoblasts is inhibited by -D-mannopyranosides or -D-glucopyranosides. After 1 hr incubation of cells with con A over 80% of the bound con A can be washed off with methyl--D-mannoside; after 24 hr only about 25% of the cell associated con A can be removed by competitor. This sequestration is temperature dependent. Cells which have been treated with con A for 20 hr still have as many combining sites for freshly added con A as do non-pretreated cells, even though about 75% of the previously added con A remains associated with the cells. Con A stimulated cells can be separated from non-stimulated cells on Ficoll gradients. Irreversibly bound con A is preferentially accumulated in the stimulated cells. However, this irreversibly bound con A does not stimulate. We suggest that irreversibly bound con A is inside the cell and that internalization is a result of the early activation of energy generating reactions by the mitogenic stimulus. The stimulatory effect depends on reactions at the cell surface.

Footnotes

¹ This work was supported, in part, by grants from the United States Atomic Energy Commission (AT(11-1)2040) and from the Leukemia Research Foundation of Chicago.

² R.M.P. is supported by United States Public Health Service Training Grant 5T05 GM0193905.

³ P.H. is supported by United States Public Health Service Training Grant GM00090-15.

⁴ E.H. is supported by United States Public Health Service Training Grant GM00603-12.

⁵ A.N. is supported by United States Public Health Service Training Grant AI00238-10.