Dobbelaere D A, Prospero T D, Roditi I J, Kelke C, Baumann I, Eichhorn M, Williams R O, Ahmed J S, Baldwin C L, Clevers H
Kernforschungszentrum Karlsruhe, Institut für Genetik und Toxikologie, Federal Republic of Germany.
Infect Immun. 1990 Dec;58(12):3847-55. doi: 10.1128/iai.58.12.3847-3855.1990.
The Tac antigen component of the bovine interleukin-2 receptor was expressed as a Cro-beta-galactosidase fusion protein in Escherichia coli and used to raise antibodies in rabbits. These antibodies were used for flow cytofluorimetric analysis to investigate the expression of Tac antigen in a variety of Theileria parva-infected cell lines and also in three Theileria annulata-infected cell lines. Cells expressing Tac antigen on their surface were found in all T. parva-infected cell lines tested whether these were of T- or B-cell origin. T cells expressing Tac antigen could be CD4- CD8-, CD4+ CD8-, CD4- CD8+, or CD4+ CD8+. Tac antigen expression was observed both in cultures which had been maintained in the laboratory for several years and in transformed cell lines which had recently been established by infection of lymphocytes in vitro with T. parva. Northern (RNA) blot analysis demonstrated Tac antigen transcripts in RNA isolated from all T. parva-infected cell lines. Three T. annulata-infected cell lines which were not of T-cell origin were also tested. Two of them expressed Tac antigen on their surface. Abundant Tac antigen mRNA was detected in these T. annulata-infected cell lines, but only trace amounts were demonstrated in the third cell line, which contained very few Tac antigen-expressing cells. In all cell lines tested, whether cloned or uncloned, a proportion of the cells did not express detectable levels of Tac antigen on their surface. This was also the case for a number of other leukocyte surface markers. In addition, we showed that the interleukin-2 receptors were biologically functional, because addition of recombinant interleukin-2 to cultures stimulated cell proliferation. Recombinant interleukin-2 treatment also resulted in increased amounts of steady-state Tac antigen mRNA. The relevance of interleukin-2 receptor expression on Theileria-infected cells is discussed.
牛白细胞介素 - 2受体的Tac抗原成分在大肠杆菌中表达为Cro-β-半乳糖苷酶融合蛋白,并用于在兔体内产生抗体。这些抗体用于流式细胞荧光分析,以研究Tac抗原在多种微小泰勒虫感染的细胞系以及三种环形泰勒虫感染的细胞系中的表达情况。在所有测试的微小泰勒虫感染的细胞系中,无论其起源于T细胞还是B细胞,均发现细胞表面表达Tac抗原。表达Tac抗原的T细胞可以是CD4 - CD8 - 、CD4 + CD8 - 、CD4 - CD8 + 或CD4 + CD8 + 。在实验室中保存数年的培养物以及最近通过体外将淋巴细胞感染微小泰勒虫而建立的转化细胞系中均观察到Tac抗原表达。Northern(RNA)印迹分析表明,从所有微小泰勒虫感染的细胞系中分离的RNA中存在Tac抗原转录本。还测试了三种非T细胞起源的环形泰勒虫感染的细胞系。其中两个在其表面表达Tac抗原。在这些环形泰勒虫感染的细胞系中检测到大量Tac抗原mRNA,但在第三个细胞系中仅检测到微量,该细胞系中表达Tac抗原的细胞很少。在所有测试的细胞系中,无论是否克隆,一部分细胞在其表面未表达可检测水平的Tac抗原。许多其他白细胞表面标志物也是如此。此外,我们表明白细胞介素 - 2受体具有生物学功能,因为向培养物中添加重组白细胞介素 - 2可刺激细胞增殖。重组白细胞介素 - 2处理还导致稳态Tac抗原mRNA量增加。讨论了白细胞介素 - 2受体在泰勒虫感染细胞上表达的相关性。