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Two Types of Anti-TL (Thymus Leukemia) CTL Clones with Distinct Target Specificities: Differences in Cytotoxic Mechanisms and Accessory Molecule Requirements¹

Kunio Tsujimura^*, Toshitada Takahashi^*, Shigeru Iwase^*,, Yasue Matsudaira^*, Yoko Kaneko, Hideo Yagita^§ and Yuichi Obata^2,*

^* Laboratory of Immunology, Aichi Cancer Center Research Institute, Nagoya; Department of Chemical Hygiene and Nutrition, Faculty of Pharmaceutical Sciences, Nagoya City University, Nagoya; Department for Geriatric Research, National Institute for Longevity Sciences, Obu; and ^§ Department of Immunology, Juntendo University School of Medicine, Tokyo, Japan

TCRß CTL clones recognizing mouse thymus leukemia (TL) Ags were established and categorized into two groups: those killing any TL⁺ target cells (type I) and those killing only TL⁺ Con A blasts (type II). Cold target inhibition assays showed that the antigenic determinant(s) recognized by type II clones are expressed not only on TL⁺ Con A blasts but also on other TL⁺ target cells. The relation of the target specificity to the killing machinery and the accessory molecules involved in cytotoxicity were therefore analyzed using four representative clones selected from each type. Of the target cells tested, Fas was only expressed on Con A blasts, indicating that Fas ligand (FasL)-dependent cytotoxicity is limited to such cells. All four type II and one of four type I clones expressed FasL on the surface, while both types contained perforin in the cytoplasm. Blocking studies using neutralizing anti-FasL mAbs and concanamycin A (CMA), a selective inhibitor of the perforin pathway, suggested that type I clones kill target cells by way of perforin, while type II clones kill TL⁺ Con A blasts through FasL together with perforin. For their cytotoxicity, type I CTLs require a signal through CD8, while type II require LFA-1/ICAM-1 interactions. Type II clones also need a costimulatory signal through an unknown molecule for perforin-dependent cytotoxicity. These results taken together suggest that the difference in the target specificity of anti-TL CTL clones is due to variation in the killing machineries and the dependence on accessory molecules.

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