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: 2070007
• 负责人: MATTHEW Franklin MESCHER
• 金额: $ 15.6万
• 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/2070007
• 关键词: MHC class I antigen; Orthomyxoviridae; 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细胞受体激活溶细胞T淋巴细胞 识别与I类MHC蛋白复合的外源肽。 是 然而，很明显，CTL的有效激活还涉及 CTL上的“辅助”蛋白（包括CD 8、LFA-1和 VLA）与抗原携带细胞上的配体。 利用人工 具有明确限定和容易变化的配体组成的膜， 在界定质量和 这些相互作用对激活的定量贡献 即时响应（例如，脱粒）。 拟议的研究将扩大这一方法，以系统地评估 辅助相互作用和淋巴因子的相对作用 活化CD 8+前体CTL用于增殖和分化， 效应子CTL。 此外，激活naive与 引发的（体内）前体CTL明显不同，将进行研究 以确定这是否是由于配体相互作用的差异 需要、细胞因子需要或两者。 同种异体和 将研究自限制的肽特异性CTL应答。 已经发现带有抗原的人工膜显著地 影响CTL应答的体内产生。 抗原给药 这种形式不会导致可检测的CTL应答，但 与活的、携带抗原的细胞一起沿着给药于小鼠， 与小鼠相比， 只接受刺激细胞。 扩增并不简单地导致 增加的抗原负荷;这种效应是抗原对细胞大小的独特影响 人工膜，似乎发生在前体CTL水平 activation. 显著增加CTL应答水平的能力具有 对肿瘤和病毒性疾病治疗的影响，以及实验 检查对同源肿瘤的细胞溶解反应，以及对肿瘤的影响 生长和宿主生存，已经证明了这一潜力 approach. 在拟定项目中，体内要求 刺激/增强将与体外 上述的活化研究。 还建议进行研究， 确定携带抗原的人工膜 能够独特地调节体内应答水平。 预计 这些实验将提供深入了解参数， 影响体内CTL应答的产生，并提出了方法 用于预防和治疗疾病。