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Characterization of Interleukin 2-dependent Cytotoxic T-Cell Clones: Specificity, Cell Surface Phenotype, and Susceptibility to Blocking by Lyt Antisera¹

Memorial Sloan-Kettering Cancer Center, New York, New York 10021

² Recipient of American Cancer Society Junior Faculty Research Award. To whom requests for reprints should be addressed.

Cytotoxic T-cells were derived from the peritoneal cavity of a C57BL/6 mouse immunized with BALB/c sarcoma Meth A and from the spleens of BALB/c x C57BL/6 F₁ (hereafter called CB6F₁) mice immunized with BALB/c leukemia RL1. The cells were cultured in interleukin 2 and cloned by limiting dilution, and their specificity was determined by direct tests and competitive inhibition assays. C57BL/6 anti-Meth A effector cells recognized H-2D^d determinants. CB6F₁ anti-RL1 effector cells recognized a unique cell surface antigen of leukemia RL1. The specificity was maintained in long-term culture. The cell surface phenotype of the cloned effector cell lines as determined by absorption analysis was Thy-1.2⁺, Lyt-1.2⁺, 2.2⁺, and 3.2⁺. Cytotoxicity was blocked at the target cell level by antisera against H-2D^d, but not H-2D^k or H-2^b, and at the effector cell level by antisera against Lyt-2.2 and 3.2, but not Lyt-1.2, Ly-5.1 or Thy-1.2.

¹ Research supported by American Cancer Society Grants IM-162C, IM-310, and JFRA-39 and by USPHS Grants CA-08748 and CA-16599 awarded by the National Cancer Institute, Department of Health and Human Services.

Received 2/ 8/82. Accepted 10/ 6/82.

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