DETECTION OF HUMAN MINOR ALLOANTIGENS USING CTL LINES

使用 CTL 系检测人类次要同种抗原

• 批准号: 3132297
• 负责人: KAREN S ZIER
• 金额: $ 12.95万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1987-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3132297
• 关键词: T lymphocyte; cytolysis; density gradient ultracentrifugation; flow cytometry; gene expression; histocompatibility antigens; histocompatibility typing; human tissue; immunochemistry; minor histocompatibility loci; parainfluenza virus type 1; radiotracer; tissue /cell culture; virus antigen; virus envelope

The study of the response by cytotoxic T lymphocytes (CTL) to minor alloantigens is of interest for a number of reasons. First, it serves as a model for the recognition of tumor and viral antigens by CTL. Second, it is of clinical relevance since minor alloantigens probably serve as the target antigens of graft-versus-host (GVHD) following bone marrow transplantation (BMT) between HLA-identical sibs. Progress in studying the recognition of minor antigens by CTL has been limited, however, by several observations. First, CTL recognize minor alloantigens only in association with self-class I antigens, not as free antigen. Thus, only target cells which share certain HLA determinants with the CTL can be tested. Second, anti-minor CTL usually can only be generated with cells from in vivo immunized responders. Finally, very little is known about the molecular nature of minor antigens. We have raised CTL lines which distinguish between members of certain HLA-identical sib pairs based on the expression of minor alloantigens. However, as CTL directed against minor alloantigens recognize these determinants only in the context of certain self HLA antigens, called restriction determinants, studies designed to detect minor alloantigens are limited to those target cells which express the restriction determinant of the CTL. Minor antigens on other cells are undetectable. Furthermore, unless a given cell expresses the restriction determinants with which two different CTL lines recognize minor antigens, it is not possible to determine whether the minor alloantigen seen by CTL 1 together with HLA-X is the same or different than the minor alloantigen seen by CTL 2 together with HLA-Y. In this project we seek to circumvent these problems. First, we will integrate plasma membranes containing the appropriate HLA antigens into potential target cells which do not express these specificities naturally. This will be accomplished using the technique of reconstituted Sendai virus envelope-mediated membrane transfer, there-by rendering the cell susceptible to lysis by the CTL line if the minor antigen is expressed. The resulting target cells could then be tested for the expression of minor antigens using all available CTL lines. This will enable studies of the population genetics of these determinants to be undertaken, as well as their systematization. Finally, we will generate anti-minor CTL in a totally in vitro system to ensure a plentiful supply of anti-minor CTL lines.

细胞毒性T淋巴细胞（CTL）对次要的反应的研究 同类物质是出于多种原因而引起的。 首先，它是 CTL识别肿瘤和病毒抗原的模型。 第二，它 是临床相关性的 骨髓后移植物抗宿主（GVHD）的靶抗原 HLA相同的SIBS之间的移植（BMT）。 研究 然而，几个 观察。 首先，CTL仅识别仅关联的小型同种抗原 使用自级I抗原，不像自由抗原。 因此，只有靶细胞 可以测试与CTL共享某些HLA决定因素的哪个。 第二， 抗少量CTL通常只能由体内细胞产生 免疫反应者。 最后，对分子知之甚少 次要抗原的性质。 我们已经提出了CTL线，以区分某些成员 HLA认同的SIB对基于次要同种抗原的表达。 但是，正如CTL针对未成年人同种抗菌原子的那样认识到这些 仅在某些自我HLA抗原的背景下，决定因素称为 限制决定因素，旨在检测次要同种抗原的研究是 限于表达限制决定因素的目标细胞 CTL。 其他细胞上的次要抗原是无法检测的。 此外， 除非给定单元表示限制决定因素 不同的CTL线识别次要抗原，不可能 确定CTL 1与HLA-X一起观察到的未成年人同种抗原 与CTL 2一起观察到的小型同种抗原相同或不同 与HLA-Y。 在这个项目中，我们试图解决这些问题。 首先，我们会的 将含有适当HLA抗原的质膜整合到 潜在的靶细胞自然表达这些特异性。 这将使用重构的仙台病毒的技术来完成 信封介导的膜转移，从而渲染细胞 如果表达次要抗原，则易于CTL线裂解。 然后可以测试所得的目标细胞的次要表达 使用所有可用CTL线的抗原。 这将使对 这些决定因素的人口遗传学以及 他们的系统化。 最后，我们将在A中生成抗少量CTL 完全有体外系统，以确保大量的抗少量CTL供应 线。