Pandolfi F, Strong D M, Slease R B, Bonnard G D
Clin Exp Immunol. 1981 Feb;43(2):319-28.
Two different specific anti-human T cell sera were studied. One was raised against fetal thymocytes (anti-HTY) and the other against human cultured T cells (anti-CTC) grown in the presence of conditioned medium from phytohaemagglutinin-stimulated peripheral blood mononuclear leucocytes (PBL). These sera, after various absorptions, reacted in microcytotoxicity assays against T cells, but not with B and null cells in the peripheral blood of both normal donors and patients with non-T leukaemias. A clear distinction between the labelling density of T versus B cells was also documented with the fluorescence-activated cell-sorter (FACS) analyses. These two sera were further absorbed on T cells from different sources with the aim of obtaining reagents specific for T cell subsets. Two reagents resulting from these absorptions were found to react with subsets of T-PBL. (1) Anti-HTY, after absorption with cells of a T-chronic lymphocyte leukaemia (T-CLL) expressing receptors for the Fc portion of IgG, reacted with thymocytes, 40–50% of normal T-PBL and only with those T cells that were not inhibited by theophylline in their ability to rosette with sheep erythrocytes. When PBL were preincubated with the absorbed serum and complement, the remaining cells had markedly diminished lymphoproliferative responses to lectins and in mixed lymphocyte culture (MLC), but they still responded well to tetanus toxoid (t.tox.). (2) Anti-CTC, after absorption with the MOLT-4 cell line reacted with about 30% of T-PBL and with the majority of the T-CLL cells with Fc receptors, as documented by cytotoxicity and FACS analyses. When normal PBL were preincubated with this absorbed serum and complement, the remaining cells had enhanced lymphoproliferative responses to lectins and especially to t.tox. and in MLC. Thus, these antisera, obtained following some rather simple absorption steps, were able to divide human T cells into two major and distinct subpopulations.
研究了两种不同的特异性抗人T细胞血清。一种是针对胎儿胸腺细胞产生的(抗-HTY),另一种是针对在植物血凝素刺激的外周血单个核白细胞(PBL)的条件培养基存在下培养的人T细胞(抗-CTC)产生的。这些血清经过各种吸收处理后,在微量细胞毒性试验中与T细胞发生反应,但在正常供体和非T白血病患者的外周血中不与B细胞和裸细胞发生反应。荧光激活细胞分选仪(FACS)分析也证明了T细胞与B细胞标记密度之间的明显差异。为了获得针对T细胞亚群的特异性试剂,这两种血清进一步用来自不同来源的T细胞进行吸收。发现由这些吸收产生的两种试剂与T-PBL的亚群发生反应。(1)抗-HTY在用表达IgG Fc部分受体的T慢性淋巴细胞白血病(T-CLL)细胞吸收后,与胸腺细胞、40%-50%的正常T-PBL发生反应,并且仅与那些在与绵羊红细胞形成玫瑰花结的能力上不受茶碱抑制的T细胞发生反应。当PBL与吸收后的血清和补体预孵育时,剩余细胞对凝集素和混合淋巴细胞培养(MLC)中的增殖反应明显减弱,但它们对破伤风类毒素(t.tox.)仍有良好反应。(2)抗-CTC在用MOLT-4细胞系吸收后,如细胞毒性和FACS分析所示,与约30%的T-PBL以及大多数具有Fc受体的T-CLL细胞发生反应。当正常PBL与这种吸收后的血清和补体预孵育时,剩余细胞对凝集素,尤其是对t.tox.以及在MLC中的增殖反应增强。因此,经过一些相当简单的吸收步骤获得的这些抗血清能够将人T细胞分为两个主要且不同的亚群。