BIOLOGY OF HUMAN TUMOR INFILTRATING LYMPHOCYTES

这项研究建议测试的假设，识别的 原位激活肿瘤浸润的要求 淋巴细胞（TIL）将提供生物学基础 在不输注离体的人实体瘤的免疫治疗中， 活化淋巴细胞 具体目标如下：1. 黑色素瘤、肾癌和肉瘤的TIL的特征。 存在至少六种不同的TIL亚群，如下： T细胞与（i）自体肿瘤特异性或（ii）MHC-T细胞结合。 非限制性（LAK）细胞毒性; CD 4 + CD 8- T细胞与（iii） 自体肿瘤特异性活性或（iv）LAK活性，（v） 具有LAK活性的CD 3-CD 16 + NK细胞和（vi）CD 4-CD 8- T细胞。 我将在每一个人之前和之后， 与rIL 2孵育;（ii）建立克隆细胞;（iii） （四）明确要求; 每个TIL亚群（IL-2、其他细胞因子、自体 肿瘤细胞或自体巨噬细胞），和（v）研究 参与TIL活化的分子（TCR，CD 3，CD 2，CD 4，CD 8， TIL上的CD 16和LFA抗原）。 2.抗肿瘤的MHC限制性 活动 我们将研究自体肿瘤特异性CTL或 黑色素瘤TIL中的活性受到MHC I类或MHC II类的限制。 通过测定肿瘤靶点上的II抗原表达， 肿瘤的MHC单倍型。3和4要求的确定 CD 4、CD 8- TIL产生IL-2， 血淋巴细胞转化为TIL。 巨噬细胞的作用。 要求 测试的包括：IL-1、IL-2、IL 2 R诱导因子、抗CD 3 mAb， 表达抗CD 3 mAb的杂交瘤，自体单核细胞，肿瘤 细胞，CD 4 + CD 8-或CD 4-CD 8+克隆的TIL。 5.抗肿瘤活性 淋巴细胞从淋巴结转移与黑色素瘤。 淋巴细胞 亚群将在IL-2，抗CD 3 mAb， IL-1和/或自体单核细胞，然后测试它们的增殖活性。 发酵和抗肿瘤活性。 6.识别和 IL-2受体诱导因子（IL 2 R-IF）的纯化。 文化 来自建立的⑶ 4 + TIL系（TILh-Ll）和PBMC的上清液 本研究将使用PHA和PMA刺激。 了解TIL的免疫学特性， TIL活化机制与宿主肿瘤生物学 关系将为发展提供重要的发现 原位淋巴细胞激活和新的策略， 人实体癌的免疫治疗。

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.