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Characterization of T and B Antigen-Binding Cells for -Galactosidase

I. -Galactosidase-Binding Cells in the Thymus and Spleen of Normal Mice^1,2,

From the Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115

Abstract

The results reported in this paper demonstrate that the enumeration of cells binding -galactosidase (Z) as an antigen, revealed by subsequent substrate hydrolysis, is an excellent method for the detection and study of antigen-binding cells (ZBC). The binding found is specific and is restricted to a small number of lymphocytes that bind a large number of Z molecules via surface receptors. Such ZBC were found at mean frequencies of 150 per 10⁶ in the thymus and 200 to 300 per 10⁶ in the spleen. The binding cells of both organs were heterogenous with individual ZBC binding from 10⁵ to 10⁶ molecules of enzyme as determined by substrate hydrolysis, although this might well be an overestimate of the number of actual receptors. The profiles for the frequency of ZBC binding different numbers of molecules were nearly identical for thymus and spleen, in contrast to descriptions of the binding of many other antigens. Receptors responsible for Z binding appear to be superficially located on the cell since they are trypsin-sensitive to a large extent and are not increased by fixation.

Footnotes

¹ Based on a thesis submitted by S. L. Swain in partial fulfillment of the requirements for the degree of Doctor of Philosophy, Harvard University. The work was supported in part by Grant AI 05691 from the National Institutes of Health and (S.S.) Training Grant 5T-01-AI 00387.

² Presented in part at the annual meeting of the Federated Societies for Experimental Biology, April, 1974 (Fed. Proc. 33:802).

³ Present address: Dept. of Biology, University of California at San Diego, La Jolla, California.

⁴ Present address: Dept. of Pathobiology, School of Public Health, Teheran University, Teheran, Iran.

⁵ Career Investigator, American Heart Association. Please send reprint requests to him.