CD4-CD8- HUMAN T-CELLS - PHENOTYPIC HETEROGENEITY AND ACTIVATION REQUIREMENTS OF FRESHLY ISOLATED DOUBLE-NEGATIVE T-CELLS

被引:26
作者
BENDER, A [1 ]
KABELITZ, D [1 ]
机构
[1] UNIV HEIDELBERG,INST IMMUNOL,NEUENHEIMER FELD 305,W-6900 HEIDELBERG,GERMANY
关键词
D O I
10.1016/0008-8749(90)90047-U
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
In the present study we have analyzed the in vitro activation requirements of freshly isolated CD4-CD8- "double-negative" (DN) human peripheral blood T cells. DN cells were isolated from E+ cells by removal of CD4+, CD8+, and CD16+ cells through consecutive steps of C-mediated lysis and panning. While the majority (79.0 ± 12.0%) of DN cells were TCRγδ+ as shown by staining with mAb TCRδ-1, a minor fraction (6.7 ± 4.7%) expressed TCR αβ as revealed by staining with mAb BMA031. Within the γδ+ DN fraction, most cells reacted with mAb TiγA which delineates a Vγ9JpCγ1 epitope, whereas a minor fraction stained with mAb δTCS-1 which identifies a Vδ1Jδ1 epitope. Functional studies performed at low cell number (1000) per microculture indicated that DN cells can be activated by anti-CD3 mAb, PHA and allogeneic stimulator cells, provided that exogenous growth factors are supplied. Both rIl-2 and rIl-4 acted as efficient growth factors for DN cells, and a synergistic stimulatory effect of rIl-2 and rIl-4 was observed when DN cells were cocultured with allogeneic LCL stimulator cells. As compared to unseparated E+ cells, isolated DN responder cells had a reduced capacity to secrete Il-2 upon PHA stimulation in the presence of LCL feeder cells. The majority of DN cells maintained their CD3+ CD4-CD8- phenotype upon coculture with allogeneic LCL stimulator cells. These data demonstrate that CD3+ DN cells in human peripheral blood are heterogeneous with respect to TCR expression. In addition, they show that freshly isolated DN cells are deficient in Il-2 production but may be normally stimulated by anti-CD3, PHA, or alloantigen if exogenous growth factors (rIl-2 and/or rIl-4) are provided. © 1990.
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页码:542 / 554
页数:13
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