CYTOTOXIC T-LYMPHOCYTE ACTIVATION IN VITRO AND IN VIVO

体外和体内细胞毒性 T 淋巴细胞激活

• 批准号: 3149640
• 负责人: MATTHEW Franklin MESCHER
• 金额: $ 14.77万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-07-01 至 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3149640
• 关键词: MHC class I antigen; T cell receptor; antigen presentation; antigen presenting cell; artificial membranes; cell adhesion molecules; cell differentiation; cell mediated lymphocytolysis test; chromium; cytotoxic T lymphocyte; immunologic memory; interferon gamma; interleukin 1; interleukin 2; interleukin 4; interleukin 6; laboratory mouse; leukocyte activation /transformation; radiotracer; tissue /cell culture; tritium; virus antigen

Cytolytic T lymphocytes are activated as a result of T cell receptor recognition of foreign peptide complexed to class I MHC proteins. It is clear, however, that effective activation of the CTL also involves interactions of 'accessory' proteins on the CTL (including CD8, LFA-1 and VLA's) with ligands on the antigen-bearing cell. Using artificial membranes having well defined and easily varied ligand compositions, considerable progress has been made in defining the qualitative and quantitative contributions that these interactions can make to activation of immediate responses (eg. degranulation) by cloned effector CTL. Proposed studies will extend this approach to systematically assess the relative roles of the accessory interactions and lymphokines in activating CD8+ precursor CTL for proliferation and differentiation to effector CTL. In addition, the requirements for activating naive versus primed (in vivo) precursor CTL clearly differ, and studies will be done to determine if this results from differences in ligand interaction requirements, cytokine requirements, or both. Both allogeneic and self-restricted, peptide specific CTL responses will be studied. Antigen-bearing artificial membranes have been found to significantly affect in vivo generation of CTL responses. Administration of antigen in this form does not result in a detectable CTL response, but administration to mice along with live, antigen-bearing cells results in a dramatic augmentation of the response compared to that obtained in mice receiving only stimulator cells. Augmentation does not simply result from increased antigen load; the effect is unique to antigen on cell-size artificial membranes and appears to occur at the level of precursor CTL activation. The ability to significantly augment CTL response levels has implications for tumor and virus disease therapy, and experiments examining cytolytic responses to syngeneic tumors, and effects on tumor growth and host survival, have demonstrated the potential of this approach. In the proposed project, the requirements for in vivo stimulation/augmentation will be studied in parallel with the in vitro activation studies described above. Studies are also proposed to determine the mechanism(s) by which antigen-bearing artificial membranes are able to uniquely modulate in vivo response levels. It is anticipated that these experiments will provide insight into the parameters which influence the in vivo generation of CTL responses, and suggest approaches for prevention and therapy of diseases.

T细胞受体激活细胞溶解T淋巴细胞 识别与I类MHC蛋白复合的外源多肽。它是 然而，很明显，有效激活CTL还包括 辅助蛋白在CTL上的相互作用(包括CD8、LFA-1和 VLA)与抗原承载细胞上的配体。使用人工的 具有定义良好且容易变化的配基组合物的膜， 在定义定性和定量的 这些相互作用对激活的数量贡献 即时响应的比例(例如通过克隆的效应器CTL。 拟议的研究将扩展这一方法，以系统地评估 辅助性相互作用和淋巴因子的相对作用 激活CD8+前体CTL增殖分化为 效应器CTL。此外，激活naive的要求与 预置的(体内)前体CTL明显不同，将进行研究 确定这是否是由于配体相互作用的差异造成的 要求，细胞因子要求，或两者兼而有之。无论是同种异体还是 我们将研究自身限制性、多肽特异性的CTL反应。 携带抗原的人工膜已经被发现显著地 影响体内CTL反应的产生。抗原在体内的注射 这种形式不会导致可检测到的CTL响应，但是 与活的携带抗原的细胞一起给小鼠注射会导致 与在小鼠身上获得的反应相比，反应显著增强 只接受刺激细胞。增强并不是简单地导致 由于抗原负荷量的增加；对细胞大小的影响是抗原独有的 人工膜，似乎发生在前体CTL的水平上 激活。显著增强CTL反应水平的能力具有 对肿瘤和病毒疾病治疗和实验的影响 检测对同基因肿瘤的细胞溶解反应及其对肿瘤的影响 生长和寄主存活，已经证明了这一潜力 接近。在拟议的项目中，体内试验的要求 刺激/增强将在体外研究的同时进行。 上述活化研究。还建议进行研究，以 确定承载抗原的人工膜的机制(S) 能够独特地调节体内的反应水平。这是意料之中的 这些实验将提供对这些参数的洞察 影响体内CTL反应的产生，并提出方法建议 用于疾病的预防和治疗。