BIOLOGY OF HUMAN TUMOR INFILTRATING LYMPHOCYTES

点击翻译按钮获取中文摘要

This research proposal tests the hypothesis that identification of the requirements for in situ activation of tumor-infiltrating lymphocytes (TIL) will provide the biological basis for immunotherapy of human solid tumors without infusion of ex vivo- activated lymphocytes. Specific aims are as follows: 1. Characterization of TIL from melanoma, renal carcinoma and sarcoma. There are at least six different TIL subsets as follows: CD4-CD8+ T cells with (i) autologous tumor-specific or (ii) MHC- nonrestricted (LAK) cytotoxicity; CD4+CD8- T cells with (iii) autologous tumor-specific The activity or (iv) LAK activity, (v) CD3-CD16+ NK cells with LAK activity and (vi) CD4-CD8- T cells. We will (i) purify each of these subsets before and after incubation with rIL2; (ii) establish cloned cells; (iii) investigate their functions; (iv) elucidate requirements for activation of each TIL subset (IL-2,other cytokines, autologous tumor cells or autologous macrophages), and (v) investigate molecules involved in the TIL activation (TCR, CD3, CD2, CD4, CD8, CD16 and LFA antigens on TIL). 2. MHC-restriction of antitumor activity. We will investigate if autologous tumor-specific CTL or The activity in melanoma TIL is restricted by MHC-Class I or Class II antigen expression on tumor targets respectively by determining MHC-haplotypes of tumors. 3 and 4. Determination of requirements for CD4 CD8- TIL to produce IL-2, and differentiation of resting blood lymphocytes into TIL. Role of macrophages. Requirements tested include: IL-l, IL-2, IL2R-inducing factor, anti-CD3 mAb., hybridomas expressing anti-CD3 mAb, autologous monocytes, tumor cells, CD4+CD8- or CD4-CD8+ cloned TIL. 5. Antitumor activity of lymphocytes from LN with melanoma metastasis. LN-lymphocyte subsets will be incubated in the presence of IL-2, anti-CD3 mAb, IL-l and/or autologous monocytes, followed by testing their proli- feration and antitumor activity. 6. Identification and purification of IL-2 receptor- inducing factor (IL2R-IF). Culture supernatants from the established CD4+ TIL line (TILh-Ll) and PBMC stimulated with PHA and PMA will be used for this study. Knowledge of immunological properties of TIL and understanding the mechanisms involved in TIL activation and the biology of host-tumor relationships will provide important findings for development of in situ lymphocyte activation and new strategies in the immunotherapy of human solid cancers.