DETECTION OF HUMAN MINOR ALLOANTIGENS USING CTL LINES

细胞毒性T淋巴细胞(CTL)对Minor免疫应答的研究 同种异体抗原之所以令人感兴趣，原因有很多。首先，它是一种 CTL识别肿瘤和病毒抗原的模型。第二，它 具有临床意义，因为较小的同种异体抗原可能作为 骨髓移植后移植物抗宿主靶抗原的研究 人类白细胞抗原相合同胞间的移植。生物信息学的研究进展 然而，CTL对次要抗原的识别受到以下几种因素的限制 观察。首先，CTL只能识别相关的次要同种异体抗原 用自身的I类抗原，而不是作为游离抗原。因此，只有目标单元格 与CTL有某些共同决定簇的细胞可以进行检测。第二, 抗微小CTL通常只能由体内的细胞产生 免疫应答者。最后，人们对这种分子知之甚少 次要抗原的性质。 我们已经提出了CTL行，以区分某些成员 基于少量同种异体抗原表达的人类白细胞抗原完全相同的同胞对。 然而，由于针对次要同种异体抗原的CTL识别这些 仅在某些自身HLA抗原的背景下的决定因素，称为 限制决定因素，旨在检测次要同种异体抗原的研究 仅限于表达的限制性决定簇的靶细胞 CTL。其他细胞上的次要抗原是检测不到的。此外， 除非给定的单元格表示两个限制行列式 不同的CTL株识别次要抗原是不可能的 确定CTL-1与人类白细胞抗原-X共同发现的次要同种异体抗原 是否与CTL 2共同看到的次要同种异体抗原相同或不同 与人类白细胞抗原Y同源性。 在这个项目中，我们试图绕过这些问题。首先，我们将 将含有适当的人类白细胞抗原的质膜整合到 不自然表达这些特异性的潜在靶细胞。 这将使用重组的仙台病毒技术来完成 包膜介导的膜转移，通过呈现细胞 如果表达了次要抗原，则容易被CTL系裂解。 然后，可以测试得到的目标细胞是否表达Minor 使用所有可用的CTL品系的抗原。这将使我们能够研究 对这些决定因素进行的群体遗传学研究，以及 他们的系统化。最后，我们将在一个 完全体外系统，以确保充足的抗微小CTL供应 台词。

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.