In vivo application of mAb directed against the γδ TCR does not deplete but generates "invisible" γδ T cells
In vivo application of mAb directed against the γδ TCR does not deplete but generates "invisible" γδ T cells
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DOI：
10.1002/eji.200838741
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发表时间：
2009-02-01
影响因子：
5.4
通讯作者：
Prinz, Immo
中科院分区：
文献类型：
--
作者：
Koenecke, Christian;Chennupati, Vijaykumar;Prinz, Immo
靶向γ δ TCR的mAb已用于γ δ T细胞耗竭，并取得了不同的成功。尽管抗γ δ TCR mAb克隆GL 3的耗竭能力一直存在争议，但许多研究组继续使用涉及mAb克隆UC 7 - 13 D5的γ δ T细胞耗竭方案，并发现了显著的生物学效应。我们在此表明，用GL 3和UC 7 - 13 D5抗体两者治疗不会在体内耗尽γ δ T细胞，而是导致TCR内化，从而产生“不可见的”γ δ T细胞。我们通过将抗γ δ TCR mAb注射到WT小鼠以及报告TCR δ基因座-组蛋白2B增强的GFP敲入小鼠中来解决这个问题，其中可以基于固有的绿色荧光检测γ δ T细胞。重要的是，使用TCR δ基因座-组蛋白2B增强的GFP小鼠首次提供了直接证据，表明“耗尽的”γ δ T细胞实际上仍然存在。我们的结果进一步表明，GL 3和UC 7 - 13 D5 mAb部分交叉竞争相同的表位。通过活化标志物评估，我们观察了通过mAb的γ δ T细胞的体外和体内活化。我们的结论是，γ δ T细胞耗竭实验必须谨慎评估，并讨论了对γ δ T细胞的生理功能的未来研究的影响。
mAb targeting the gamma delta TCR have been used for gamma delta T-cell depletion with varying success. Although the depletion-capacity of the anti-gamma delta TCR mAb clone GL3 has been disputed repeatedly, many groups continue to use gamma delta T-cell depletion protocols involving the mAb clone UC7-13D5 and find significant biological effects. We show here that treatment with both GL3 and UC7-13D5 antibodies does not deplete gamma delta T cells in vivo, but rather leads to TCR internalization and thereby generates "invisible" gamma delta T cells. We addressed this issue using anti-gamma delta TCR mAb injections into WT mice as well as into reporter TCR delta locus-histone 2B enhanced GFP knock-in mice, in which gamma delta T cells can be detected based on an intrinsic green fluorescence. Importantly, the use of TCR delta locus-histone 2B enhanced GFP mice provided here for the first time direct evidence that the "depleted" gamma delta T cells were actually still present. Our results show further that GL3 and UC7-13D5 mAb are in part cross-competing for the same epitope. Assessed by activation markers, we observed in vitro and in vivo activation of gamma delta T cells through mAb. We conclude that gamma delta T-cell depletion experiments must be evaluated with caution and discuss the implications for future studies on the physiological functions of gamma delta T cells.