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Cell-Mediated Immune Responses in Vitro

II. Simultaneous Generation of Cytotoxic Lymphocyte Responses to Two Sets of Alloantigens of Limited Cross-Reactivity¹

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

Abstract

The conditions for generation of simultaneous and independent cytotoxic lymphocyte (CL) responses to each of two sets of alloantigens of limited cross-reactivity by mouse spleen cells in vitro have been investigated. Responder spleen cells were incubated with mitomycin C-treated C57BL/6 (H-2^b) or DBA/2 (H-2^d) stimulator spleen cells and day 5 CL responses were assayed with ⁵¹Cr-labeled EL-4 leukemia (H-2^b) and P815 mastocytoma (H-2^d) as target cells. Spleen cells from mice of the various H-2 haplotypes tested differed greatly in their ability to develop specific CL responses against alloantigens on the stimulator spleen cells and in the degree of cross-reactive cytotoxic activity against target cells bearing alloantigens not present on the stimulator spleen cells. In contrast to the other strains examined, DBA/1 (H-2^q) spleen cells developed specific CL responses to either H-2^b or H-2^d alloantigens without exhibiting significant cross-reactive activity on the inappropriate target cell. The CL responses to H-2^b and H-2^d alloantigens by DBA/1 spleen cells were comparable in magnitude and had similar stimulator cell-dose requirements. Further, DBA/1 spleen cells developed CL responses of normal magnitude simultaneously against both target cells when incubated with both mitomycin C-treated C57BL/6 and DBA/2 stimulator cells.

Footnotes

¹ This investigation was supported by United States Public Health Service Program Project Grant CA-14723 from the National Cancer Institute.

² Recipient of United States Public Health Service Postdoctoral Fellowship CA-00566 from the National Cancer Institute.

³ Recipient of United States Public Health Service Research Career Development Award 5K4-AI-70173 from the National Institute of Allergy and Infectious Diseases.