Hinz T, Wesch D, Friese K, Reckziegel A, Arden B, Kabelitz D
Department of Immunology, Paul Ehrlich Institute, Langen, Germany.
Eur J Immunol. 1994 Dec;24(12):3044-9. doi: 10.1002/eji.1830241219.
While V gamma 9/V delta 2 cells dominate among peripheral blood gamma delta T cells in healthy adults, the majority of gamma delta T cells in most HIV-1-infected individuals express V delta 1. We asked whether these elevated levels of V delta 1 T cells were due to clonal expansion. Three-color flow cytometry with monoclonal antibodies against V gamma 2/V gamma 3/V gamma 4, V gamma 4 and V gamma 9 was used to investigate V gamma usage in 27 patients with elevated numbers of V delta 1 T cells. While the relative proportion of V gamma 9 cells among gamma delta T cells was significantly reduced in HIV-1+ individuals (10 +/- 11% vs. 80 +/- 17%, p < 0.001), the fraction of gamma delta T cells using V gamma 5 or V gamma 8 was significantly increased (54 +/- 15% vs. 7 +/- 11%, p < 0.001). In 1 patient, 76% of the V delta 1 cells expressed V gamma 2 or V gamma 3, suggesting clonality of the V delta 1 population. In line with this assumption, analysis of the V delta 1-J delta junctional regions by reverse transcription-polymerase chain reaction (RT-PCR) resulted in products of only one junctional length, as demonstrated by electrophoresis on denaturing gels, and 12 out of 16 (75%) in-frame junctional sequences were identical in this patient. In other HIV-1+ patients, RT-PCR resulted in products of several distinct sizes, also indicating a highly restricted repertoire. After sequencing the V delta 1-J delta junctional regions of 3 additional patients, we found repeated but patient-specific in-frame junctions accounting for 10-30% of the sequenced clones. However, limited V delta 1-J delta junctional diversity was also seen in healthy donors. RT-PCR products from 10 healthy individuals resulted in distinct bands on denaturing gels. In 1 of them exhibiting a single prominent band, 10 out of 17 (58%) sequenced junctions were identical. Two other healthy donors displayed 2/14 and 5/18 identical junctional sequences, respectively. Taken together, our results reveal significant alterations of V gamma usage in HIV-1+ patients, while the V delta 1 junctional repertoire is similarly restricted in HIV-1+ and HIV-1- individuals. Therefore, these data argue against an obligatory clonal expansion of V delta 1-expressing cells during HIV-1 infection.
在健康成年人外周血γδT细胞中，Vγ9/Vδ2细胞占主导地位，但在大多数HIV-1感染者中，多数γδT细胞表达Vδ1。我们探讨了这些Vδ1 T细胞水平升高是否是由于克隆扩增所致。使用针对Vγ2/Vγ3/Vγ4、Vγ4和Vγ9的单克隆抗体进行三色流式细胞术，以研究27例Vδ1 T细胞数量升高患者的Vγ使用情况。在HIV-1阳性个体中，γδT细胞中Vγ9细胞的相对比例显著降低（10±11%对80±17%，p<0.001），而使用Vγ5或Vγ8的γδT细胞比例显著增加（54±15%对7±11%，p<0.001）。在1例患者中，76%的Vδ1细胞表达Vγ2或Vγ3，提示Vδ1群体具有克隆性。与此假设一致，通过逆转录-聚合酶链反应（RT-PCR）分析Vδ1-Jδ连接区，变性凝胶电泳显示产物只有一种连接长度，该患者16个（75%）读框内连接序列中有12个相同。在其他HIV-1阳性患者中，RT-PCR产生几种不同大小的产物，也表明其谱系高度受限。对另外3例患者的Vδ1-Jδ连接区进行测序后，我们发现重复但患者特异性的读框内连接，占测序克隆的10%-30%。然而，在健康供者中也观察到Vδ1-Jδ连接区多样性有限。10名健康个体的RT-PCR产物在变性凝胶上产生不同条带。其中1例显示单一突出条带，17个测序连接中有10个（58%）相同。另外两名健康供者分别显示2/14和5/18相同的连接序列。综上所述，我们的结果揭示了HIV-1阳性患者Vγ使用情况的显著改变，而Vδ1连接区谱系在HIV-1阳性和HIV-1阴性个体中同样受限。因此，这些数据表明HIV-1感染期间表达Vδ1的细胞并非必然发生克隆扩增。
