In vivo identification of glycolipid antigen-specific T cells using fluorescent CD1d tetramers.
In vivo identification of glycolipid antigen-specific T cells using fluorescent CD1d tetramers.
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DOI：
10.1084/jem.191.11.1895
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发表时间：
2000-06-05
影响因子：
15.3
通讯作者：
Bendelac, A
中科院分区：
文献类型：
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作者：
Benlagha, K;Weiss, A;Beavis, A;Teyton, L;Bendelac, A
主要组织相容性复合体（MHC）样分子的CD1家族专门负责将脂质和糖脂抗原呈递给α/β T淋巴细胞，但对于CD1限制性T细胞群体的规模以及针对特定糖脂抗原的T淋巴细胞的频率知之甚少。在此，我们报道了小鼠CD1d1 - 糖脂四聚体的产生及其用于可视化CD1d限制性T细胞的用途。与先前基于表面等离子共振（BIAcore）对CD1d - 糖脂复合物极短半衰期的估计相反，我们发现几种不同的CD1d - 糖脂复合物的解离速率非常缓慢。小鼠CD1d1与α-半乳糖神经酰胺（αGalCer）形成的荧光四聚体（αGalCer是被小鼠Vα14 - Jα281/Vβ8和人Vα24 - JαQ/Vβ11自然杀伤T（NKT）细胞T细胞受体（TCRs）识别的抗原）使我们首次能够基于TCR特异性准确描述体内整个NKT细胞群体，并鉴定出一个先前未被识别的NK1.1阴性的“NKT”细胞群体，该群体表达一种不同的整合素模式。相比之下，自然杀伤（NK）细胞不能结合空载或负载αGalCer的四聚体，这表明不存在CD1d特异性、抗原非特异性的NK受体。小鼠CD1d1 - αGalCer四聚体也能对人NKT细胞染色，这表明它们将有助于探究一系列小鼠和人类疾病，如胰岛素依赖型糖尿病、肿瘤排斥以及NKT细胞起重要作用的感染性疾病。
The CD1 family of major histocompatibility complex (MHC)-like molecules specializes in presenting lipid and glycolipid antigens to α/β T lymphocytes, but little is known about the size of the CD1-restricted T cell population or the frequency of T lymphocytes specific for a given glycolipid antigen. Here, we report the generation and use of mouse CD1d1–glycolipid tetramers to visualize CD1d-restricted T cells. In contrast with previous BIAcore-based estimates of very short half-lives for CD1d–glycolipid complexes, we found that the dissociation rate of several different CD1d–glycolipid complexes was very slow. Fluorescent tetramers of mouse CD1d1 complexed with α-galactosylceramide (αGalCer), the antigen recognized by mouse Vα14-Jα281/Vβ8 and human Vα24-JαQ/Vβ11 natural killer T (NKT) cell T cell receptors (TCRs), allowed us for the first time to accurately describe, based on TCR specificity, the entire population of NKT cells in vivo and to identify a previously unrecognized population of NK1.1-negative “NKT” cells, which expressed a different pattern of integrins. In contrast, natural killer (NK) cells failed to bind the tetramers either empty or loaded with αGalCer, suggesting the absence of a CD1d-specific, antigen-nonspecific NK receptor. Mouse CD1d1–αGalCer tetramers also stained human NKT cells, indicating that they will be useful for probing a range of mouse and human conditions such as insulin-dependent diabetes mellitus, tumor rejection, and infectious diseases where NKT cells play an important role.