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.
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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DOI：
10.1084/jem.168.2.491
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发表时间：
1988-08-01
期刊：
影响因子：
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通讯作者：
Moretta A
中科院分区：
文献类型：
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作者：
Bottino C;Tambussi G;Ferrini S;Ciccone E;Varese P;Mingari MC;Moretta L;Moretta A
分析了两种针对 TCR-gamma/delta 的 mAb 与 CD3+WT31- 细胞群或克隆的反应模式。在正常个体中，BB3 mAb 与大约 2/3 的外周血 CD3+WT31- 淋巴细胞发生反应，而 delta-TCS-1 则对大约 1/3 的此类细胞进行染色。此外，BB3+ 和 delta-TCS-1+ 细胞的百分比之和近似于所测试的 7 名正常供体中外周血 CD3+WT31- 淋巴细胞的百分比。此外，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，在未分级和 CD4-8 胸腺细胞群中只能检测到 delta-TCS-1 反应性细胞，而不能检测到 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 发生反应，但不与 delta-TCS-1 mAb 发生反应。另一方面，delta-TCS-1 mAb 对剩余的 BB3-克隆进行染色。表达中低量CD8抗原的五个克隆是BB3-δ-TCS-1+。两种类型的克隆均在抗 CD3 mAb（但不存在针对 HLA-DR、CD7 分子或 TCR-α/β 的 mAb）存在下裂解带有 Fc γ 受体的 P815 靶细胞。在此溶细胞测定中，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。在细胞表面碘化和用相应的抗TCR mAb或抗CD3 mAb（在含有毛地黄皂苷的缓冲液中）免疫沉淀后，分析由BB3+或delta-TCS-1+克隆表达的CD3相关表面分子。在 SDS-PAGE 中，从 13 个 BB3+ 克隆中免疫沉淀的分子在非还原条件下显示出 80 kD 的分子量（在某些情况下，可以检测到较小的 38 kD 带）。在还原条件下，检测到 44 和 41 kD 的两个主要成分（以及 38 kD 的次要成分）。另一方面，在还原和非还原条件下，来自 11 个不同 delta-TCS-1+ 克隆的 TCR 分子免疫沉淀显示为 41-44 kD 的弥散带（在非还原条件下，存在额外的 38 kD 带）。因此，BB3+细胞表达二硫键连接形式的TCR-γ/δ，而δ-TCS-1+细胞表达非二硫键连接形式。（摘要截断为400字）
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)