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）对小细胞肺癌细胞免疫应答的研究 同种异体抗原由于许多原因而受到关注。 首先，它作为一个 CTL识别肿瘤和病毒抗原的模型。 二是 具有临床相关性，因为次要的同种异体抗原可能充当 骨髓移植后移植物抗宿主（GVHD）靶抗原 HLA相同同胞之间的骨髓移植（BMT）。 研究的进展 然而，CTL对次要抗原的识别受到了一些限制， 意见。 首先，CTL仅识别联合的次要同种抗原 自身I类抗原，而不是游离抗原。 因此，只有靶细胞 其与CTL共享某些HLA决定簇。 第二、 抗次要CTL通常只能用来自体内的细胞产生 免疫应答者。 最后，我们对分子水平的 次要抗原的性质。 我们已经提出了CTL系，其区分某些 基于次要同种异体抗原表达的HLA相同同胞对。 然而，由于针对次要同种异体抗原的CTL识别这些抗原， 决定簇仅在某些自身HLA抗原的背景下，称为 限制性决定簇，旨在检测次要同种抗原的研究 仅限于表达限制性决定子的那些靶细胞， 的CTL。 其他细胞上的次要抗原是检测不到的。 此外，委员会认为， 除非一个给定的细胞表达了两个限制性决定簇， 不同的CTL细胞系识别次要抗原， 确定CTL 1与HLA-X一起看到的次要同种异体抗原 与CTL 2共同观察到的次要同种异体抗原相同或不同 HLA-Y 在这个项目中，我们试图规避这些问题。 一是 将含有适当HLA抗原的质膜整合到 潜在的靶细胞不表达这些特异性天然。 这将通过重组仙台病毒的技术来实现 细胞介导的膜转移，使细胞 如果表达次要抗原，则对CTL系的裂解敏感。 然后可以测试所得的靶细胞的次要蛋白质的表达。 使用所有可获得的CTL系的抗原。 这将使研究 这些决定因素的人口遗传学，以及 其系统化。 最后，我们将在一个 完全在体外系统，以确保抗次要CTL的充足供应 线