Ptak W, Szczepanik M, Ramabhadran R, Askenase P W
Department of Immunology, Jagiellonian University College of Medicine, Krakow, Poland.
J Immunol. 1996 Feb 1;156(3):976-86.
In the current study, we confirmed previous findings suggesting that gamma delta T cells were involved in the successful adoptive cell transfer of contact sensitivity (CS) by alpha beta CS-effector T cells. In this study, we used hamster anti-mouse gamma delta-TCR mAb treatment of CS-effector T cells, followed by enrichment and removal of the gamma delta T cells with goat anti-hamster Ig-linked magnetic beads, or by addition of hemolytic rabbit C. This removal of gamma delta T cells abrogated adoptive cell transfers of CS, despite the presence of alpha beta T cells that are known to mediate CS. FACS analysis documented enrichment of gamma delta T cells rising from 1 to 2% of the starting cells, to 60 to 95% of the magnetic bead adherent cells. Adoptive cell transfer of CS was reconstituted by adding back to the alpha beta cells, highly enriched gamma delta cells attached to anti-gamma delta-TCR magnetic beads. Not only were gamma delta-enriched T cells from sensitized mice able to assist immune CS-effector alpha beta T cells, but gamma delta T cells from normal nonimmune mice also had CS-assisting activity, and furthermore, neither were MHC-restricted in this function. Thus, CS-assisting gamma delta T cells were present endogenously in normal mice without prior immunization, and acted without Ag specificity and without MHC restriction, to assist CS-effector alpha beta T cells. Similar studies, with hamster mAbs specific for V gamma and V delta portions of gamma delta-TCR, demonstrated that the gamma delta T cells that assisted the CS-effector alpha beta T cells preferentially expressed V gamma 5 and V delta 4 in their TCR. PCR analysis on extracted mRNA showed that V gamma 5 and V delta 4 gene segments indeed were rearranged and expressed in the sensitized and normal lymph nodes; and one-and two-color FACS analysis of magnetic bead-fractionated cells suggested that V gamma 5 and V delta 4 were expressed on the same T cells. In summary, these results demonstrated that V gamma 5+, V delta 4+, gamma delta T cells were needed to assist alpha beta effector T cells in the adoptive cell transfer of CS.
在当前研究中，我们证实了先前的研究结果，即γδT细胞参与了αβ接触敏感性（CS）效应T细胞成功的接触敏感性过继性细胞转移。在本研究中，我们用仓鼠抗小鼠γδ-TCR单克隆抗体处理CS效应T细胞，随后用山羊抗仓鼠Ig连接的磁珠富集并去除γδT细胞，或加入溶血性兔补体C。尽管存在已知介导CS的αβT细胞，但去除γδT细胞仍废除了CS的过继性细胞转移。流式细胞术分析表明，γδT细胞从起始细胞的1%至2%富集到磁珠粘附细胞的60%至95%。通过将附着于抗γδ-TCR磁珠的高度富集的γδ细胞重新添加到αβ细胞中，可重建CS的过继性细胞转移。不仅来自致敏小鼠的富含γδ的T细胞能够辅助免疫CS效应αβT细胞，而且来自正常非免疫小鼠的γδT细胞也具有CS辅助活性，此外，该功能均不受MHC限制。因此，CS辅助性γδT细胞在正常小鼠中内源性存在，无需预先免疫，且在无抗原特异性和无MHC限制的情况下发挥作用，以辅助CS效应αβT细胞。用针对γδ-TCR的Vγ和Vδ部分的仓鼠单克隆抗体进行的类似研究表明，辅助CS效应αβT细胞的γδT细胞在其TCR中优先表达Vγ5和Vδ4。对提取的mRNA进行的PCR分析表明，Vγ5和Vδ4基因片段确实在致敏和正常淋巴结中发生重排并表达；对磁珠分离细胞进行的单双色流式细胞术分析表明，Vγ5和Vδ4在同一T细胞上表达。总之，这些结果表明，Vγ5+、Vδ4+、γδT细胞在CS的过继性细胞转移中是辅助αβ效应T细胞所必需的。
