Analysis of Mechanisms in Maturation of Human T Cells in the Thymus and Characterization of Various Factors Affecting Those Process.

胸腺中人类 T 细胞成熟机制的分析以及影响这些过程的各种因素的表征。

基本信息

批准号：
63440038
负责人：
MATSUMOTO Shuzo
金额：
$ 6.34万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

项目摘要

In order to study the mechanisms in maturation of human T cells in thymus, a part of human thymus was obtained from patients with congenital heart diseases at their operation. Using bovine serum albumin discontinuous density gradient method, thymocytes were fractionated into 5 populations. Each fraction was studied for the expression of T cell antigens, CD1, 3, 4, 8, 25, 45, and WT31 by flow-cytometry analysis. They were also cultured in the presence or absence of IL-2 or IL-4. Finally, DNA extracted from each thymocyte population were analyzed for the rearrangement of T cell antigen receptor beta and gamma chain.There were more CD3^<low+> or CD4^+CD8^+ double positive population in high density fraction than in lower density population. In contrast, there were less CD4^+CD8^- or CD4^-CD8^+ single positive population in high density fraction. In the analysis of surface T cell antigen receptor, the expressions of were in direct proportion to that of CD3 in all fractions. Although the pe … More ak of their distribution was observed at negative region in Fr. I, the peaks were obtained at both the same region and high intensity region in Fr. III. However, that in Fr. V was observed at low intensity positive region.In the induction analysis of IL-2R, the addition of PMA induced surface IL-2R on the CD3^+ cells in all fractions, although the addition of PHA or Con A did not induce on CD3^+ cells in Fr. V. In the analysis of proliferative response to mitogens, the cells in Fr. I responded to the addition of PHA or Con A although those in Fr. III did not response even with rIL-2. The addition of PMA with rIL-2 induced the proliferative response in all fraction although that without rIL-2 did not induce in any fractions.In the induction analysis of 2H4 antigen of thymocytes in Fr. III, the addition of rIL-4 alone induced 2H4 antigen only in CD3^+ population, although PMA alone did not induce in any population. Moreover, rIL-4 and PMA induced surface 2H4 antigen much intensively in CD3^+. In thymocytes in Fr. V, the additions of rIL-4 and PMA slightly induced 2H4 antigens in both of CD3^+ and CD3^- populations although rIL-4 alone did not induce anymore.Finally, in the analyses of T cell antigen receptor beta chain rearrangement, polyclonal rearrangements were observed in all fractionated thymocyte populations, suggesting non-functional rearrangements occurred in high density population. Less

为了研究胸腺中人类T细胞成熟的机制，在手术时从先天性心脏病患者那里获得了人胸部的一部分。使用牛血清白蛋白不连续的密度梯度法，将胸腺细胞分级为5个种群。通过流量仪分析，研究了每个馏分的T细胞抗原，CD1、3、4、8、25、45和WT31的表达。在存在或不存在IL-2或IL-4的情况下，它们也被培养。最后，分析从每个胸腺细胞群体中提取的DNA，以重排T细胞抗原受体β和伽马链的重排。在高密度种群中，高密度分数中的CD3^<+>或CD4^+CD8^+双重阳性人群的CD3^^<+>或CD4^+CD8^+双重阳性多。相反，在高密度分数中，CD4^+ CD8^ - 或CD4^-CD8^+单个阳性种群。在对表面T细胞抗原受体的分析中，在所有馏分中，表达与CD3的表达成正比。尽管在Fr的负区域观察到了PE…更多的分布。 i，在Fr的同一区域和高强度区域都获得了峰。 iii。但是，在神父中。在低强度阳性区域观察到V。在对IL-2R的诱导分析中，在所有馏分中，PMA诱导的表面IL-2R在CD3^+细胞上添加，尽管添加PHA或CON A不影响FR中的CD3^+细胞。 V.在分析对有丝分裂原的反应的分析中，Fr中的细胞。我回应了加入PHA或CON A，尽管那些在神父中。 III即使使用RIL-2也没有回应。使用RIL-2添加PMA诱导了所有分数中没有RIL-2在任何馏分中都不会影响的反应。 iii，仅添加RIL-4仅在CD3^+种群中诱导2H4抗原，尽管单独的PMA并未诱导任何人群。此外，RIL-4和PMA在CD3^+中非常强烈地诱导表面2H4抗原。在Fr的胸腺细胞中。 v，在CD3^+和CD3^ - 种群中，RIL-4和PMA的添加略有诱导的2H4抗原，尽管仅RIL-4不再诱导。在本文中，在T细胞抗原受体β链重排重排重排重排的分析中，在所有分数的胸膜分数中都在较高的胸膜群中观察到了较高的thymociytecys群体，建议较高的群体。较少的