Isolation and characterization of CD6- T cells from peripheral blood.
Isolation and characterization of CD6- T cells from peripheral blood.
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DOI：
10.4049/jimmunol.152.2.527
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发表时间：
1994-01
影响因子：
4.4
通讯作者：
R. Rasmussen;S. L. Counts;Jennifer Daley;S. F. Schlossman
中科院分区：
文献类型：
--
作者：
R. Rasmussen;S. L. Counts;Jennifer Daley;S. F. Schlossman
针对CD 6 Ag的抗体已被描述为具有泛T细胞反应性。然而，我们最近已经证明，在用抗CD 6阻断的蓖麻毒素结合的免疫毒素治疗PBL后，可以鉴定出CD 3+、CD 6-细胞的克隆群体。在此，我们通过新鲜分离的E玫瑰花结+细胞的双参数染色显示，平均5%至6%的CD 3+或CD 5+细胞在其表面上表达很少或不表达CD 6。在通过抗体包被的顺磁珠耗竭进行阴性选择后，扩增的CD 6- T细胞显示为CD 1a-、CD 2+、CD 3+、CD 5+、CD 16-、CD 56-、TCR-γ/δ-，并且由CD 4+和CD 8+细胞组成。此外，毛地黄皂苷透化的细胞染色显示细胞质中没有CD 6 Ag的表达，并且通过北方印迹分析未检测到CD 6 mRNA。对于通过珠耗尽或免疫毒素处理分离并用PHA或固定化抗CD 3 mAb扩增的T细胞克隆，观察到相同的染色模式。还发现，相对于未分级的T细胞，CD 6- T细胞上的CD 5的表面表达显著减少。在功能上，新鲜分离的CD 6- T细胞表现出显着降低同种异体反应性MLR相比，未分级E-玫瑰花结+细胞，但都给出了类似的增殖反应，无论是PHA或可溶性破伤风毒素Ag。我们的结论是，存在一个次要的外周血中的成熟T细胞亚群，缺乏CD 6。这些细胞的同种异体反应性降低可能有助于解释移植物抗宿主病的低发生率，尽管在接受抗CD 6（T12）mAb加C '治疗的异基因骨髓移植患者中报告了高水平的植入。
Antibodies to the CD6 Ag have been described as having pan-T cell reactivity. We have recently demonstrated, however, that after treatment of PBL with an anti-CD6-blocked ricin-conjugated immunotoxin, clonal populations of CD3+, CD6- cells can be identified. Herein we show that through dual parameter staining of freshly isolated E-rosette+ cells, an average of 5 to 6% of either CD3+ or CD5+ cells express little or no CD6 on their surface. After negative selection by antibody-coated paramagnetic bead depletion, expanded CD6- T cells were shown to be CD1a-, CD2+, CD3+, CD5+, CD16-, CD56-, TCR-gamma/delta-, and consisted of both CD4+ and CD8+ cells. Furthermore, staining of digitonin permeabilized cells showed no cytoplasmic expression of the CD6 Ag and CD6 mRNA was not detected by Northern blot analysis. Identical staining patterns were observed for T cell clones isolated through bead depletion or immunotoxin treatment and expanded with either PHA or immobilized anti-CD3 mAb. It was also found that, relative to unfractionated T cells, the surface expression of CD5 was significantly diminished on CD6- T cells. Functionally, freshly isolated CD6- T cells showed substantially reduced alloreactivity in MLR compared with unfractionated E-rosette+ cells, yet both gave similar proliferative responses to either PHA or soluble tetanus toxin Ag. We conclude that there exists a minor subpopulation of mature T cells in peripheral blood that lack CD6. The diminished alloreactivity of these cells may help to explain the low incidence of graft-vs-host disease, despite high levels of engraftment, that has been reported in allogeneic bone marrow transplant patients receiving marrow treated with anti-CD6 (T12) mAb plus C'.