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首页> 外文期刊>The Journal of Experomental Medicine >Two subsets of human T lymphocytes expressing gamma/delta antigen receptor are identifiable by monoclonal antibodies directed to two distinct molecular forms of the receptor.
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Two subsets of human T lymphocytes expressing gamma/delta antigen receptor are identifiable by monoclonal antibodies directed to two distinct molecular forms of the receptor.

机译:表达γ/δ抗原受体的人T淋巴细胞的两个子集可以通过针对受体的两种不同分子形式的单克隆抗体来鉴定。

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Two mAbs directed to the TCR-gamma/delta were analyzed for their pattern of reactivity with CD3+WT31- cell populations or clones. In normal individuals, the BB3 mAb reacted with approximately 2/3 of peripheral blood CD3+WT31- lymphocytes, whereas delta-TCS-1 stained approximately 1/3 of such cells. In addition, the sum of the percentages of BB3+ and delta-TCS-1+ cells approximated the percentages of peripheral blood CD3+WT31- lymphocytes in seven normal donors tested. Also, in peripheral blood-derived polyclonal CD3+WT31- populations, cultured in IL-2, cells reacting with one or another mAb accounted for the whole cell population. On the other hand, only delta-TCS-1-reactive cells, but not BB3+ cells, could be detected in unfractionated as well as in CD4-8-thymocyte populations. Analysis of peripheral blood-derived CD3+WT31- clones showed that 70% of 72 clones analyzed reacted with BB3 mAb, but not with delta-TCS-1 mAb. On the other hand, delta-TCS-1 mAb stained the remaining BB3- clones. Five clones expressing medium-low amounts of CD8 antigen were BB3- delta-TCS-1+. Both types of clones lysed the Fc gamma receptor-bearing P815 target cell in the presence of anti-CD3 mAb (but not of mAb directed against HLA-DR, CD7 molecules, or TCR-alpha/beta). In this cytolytic assay, BB3 mAb induced target cell lysis only by BB3+ clones, whereas delta-TCS-1 mAb was effective only with delta-TCS-1+ clones. The CD3-associated surface molecules expressed by BB3+ or delta-TCS-1+ clones were analyzed after cell surface iodination and immunoprecipitation with the corresponding anti-TCR mAb or with anti-CD3 mAb (in digitonin-containing buffer). In SDS-PAGE, molecules immunoprecipitated from 13 BB3+ clones displayed, under nonreducing conditions, a molecular weight of 80 kD (in some cases, a minor 38-kD band could be detected). Under reducing conditions, two major components of 44 and 41 kD (and a minor component of 38 kD) were detected. On the other hand, TCR molecules immunoprecipitated from 11 different delta-TCS-1+ clones appeared as a diffuse band of 41-44 kD, both under reducing and nonreducing conditions (under non-reducing condition, an additional 38-kD band was present). Therefore, BB3+ cells express a disulphide-linked form of TCR-gamma/delta whereas delta-TCS-1+ cells express a non-disulphide-linked form.(ABSTRACT TRUNCATED AT 400 WORDS)
机译:分析了针对TCR-γ/δ的两个mAb与CD3 + WT31-细胞群体或克隆的反应模式。在正常个体中,BB3 mAb与大约2/3的外周血CD3 + WT31-淋巴细胞反应,而delta-TCS-1将大约1/3的此类细胞染色。另外,在测试的七个正常供体中,BB3 +和δ-TCS-1+细胞的百分比之和近似于外周血CD3 + WT31-淋巴细胞的百分比。另外,在IL-2中培养的外周血来源的多克隆CD3 + WT31-群体中,与一种或另一种mAb反应的细胞占整个细胞群体。另一方面,在未分级分离的以及CD4-8胸腺细胞群体中,仅可检测到δ-TCS-1-反应性细胞,而未检测到BB3 +细胞。对外周血来源的CD3 + WT31-克隆的分析表明,分析的72个克隆中有70%与BB3 mAb反应,但与delta-TCS-1 mAb不反应。另一方面,δ-TCS-1mAb染色了其余的BB3-克隆。表达中低量CD8抗原的五个克隆为BB3- delta-TCS-1 +。两种类型的克隆都在抗CD3 mAb(但不是针对HLA-DR,CD7分子或TCR-alpha / beta的mAb)存在的情况下裂解了带有Fcγ受体的P815靶细胞。在这种溶细胞测定中,BB3 mAb仅通过BB3 +克隆诱导靶细胞裂解,而delta-TCS-1 mAb仅对delta-TCS-1 +克隆有效。在细胞表面碘化并用相应的抗-TCR mAb或抗-CD3 mAb(含洋地黄毒苷的缓冲液)免疫沉淀后,分析了由BB3 +或δ-TCS-1+克隆表达的与CD3相关的表面分子。在SDS-PAGE中,在非还原条件下,从13个BB3 +克隆中免疫沉淀的分子显示出80 kD的分子量(在某些情况下,可以检测到38 kD的较小条带)。在还原条件下,检测到两个主要成分分别为44 kD和41 kD(次要成分为38 kD)。另一方面,从11种不同的delta-TCS-1 +克隆中免疫沉淀的TCR分子在还原和非还原条件下均表现为41-44 kD的扩散带(在非还原条件下,还存在一条38 kD的带) )。因此,BB3 +细胞表达TCR-γ/δ的二硫键连接形式,而δ-TCS-1+细胞表达非二硫键连接的形式(摘要截短400字)。

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