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一种在体内可视化多克隆病毒特异性CD8 T细胞的新方法。

A novel approach to visualize polyclonal virus-specific CD8 T cells in vivo.

作者信息

Zimmermann C, Pircher H

机构信息

Department of Immunology, Institute for Medical Microbiology and Hygiene, University of Freiburg, Germany.

出版信息

J Immunol. 1999 May 1;162(9):5178-82.

Abstract

Recent technical breakthroughs in generating soluble MHC class I-peptide tetramers now allow the direct visualization of virus-specific CD8 T cells after infection in vivo. However, this technique requires the knowledge of the immunodominant viral epitopes recognized by T cells. Here, we describe an alternative approach to visualize polyclonal virus-specific CD8 T cells in vivo using a simple adoptive transfer system. In our approach, C57BL/6 (Thy1.2) mice were infected with lymphocytic choriomeningitis virus, vesicular stomatitis virus, or vaccinia virus to induce virus-specific memory T cells. Tracer T cells (2 x 106) from these virus-immune mice were adoptively transferred into nonirradiated (C57BL/6 x B6.PL-Thy-1a)F1 mice. After infection of the F1-recipient mice with the appropriate virus, the transferred cells expanded vigorously, and on day 8 postinfection 60-80% of total CD8 T cells were of donor T cell origin. Under the same conditions memory CD4 T cells gave rise to at least 10 times less cell numbers than memory CD8 T cells. The transfer system described here not only allows to visualize effector and memory CD8 T cells in vivo but also to isolate them for further in vitro characterization without knowing the epitopes recognized by these Ag-specific CD8 T cells.

摘要

近期在生成可溶性Ⅰ类主要组织相容性复合体-肽四聚体方面的技术突破,使得在体内感染后能够直接观察到病毒特异性CD8 T细胞。然而,该技术需要了解T细胞识别的免疫显性病毒表位。在此,我们描述了一种使用简单的过继转移系统在体内观察多克隆病毒特异性CD8 T细胞的替代方法。在我们的方法中,用淋巴细胞性脉络丛脑膜炎病毒、水疱性口炎病毒或痘苗病毒感染C57BL/6(Thy1.2)小鼠,以诱导病毒特异性记忆T细胞。将来自这些病毒免疫小鼠的示踪T细胞(2×10⁶个)过继转移到未受照射的(C57BL/6×B6.PL-Thy-1a)F1小鼠体内。在用合适的病毒感染F1受体小鼠后,转移的细胞大量增殖,在感染后第8天,总CD8 T细胞中有60% - 80%来源于供体T细胞。在相同条件下,记忆性CD4 T细胞产生的细胞数量比记忆性CD8 T细胞至少少10倍。本文所述的转移系统不仅能够在体内观察效应性和记忆性CD8 T细胞,还能在不知道这些抗原特异性CD8 T细胞所识别表位的情况下将它们分离出来,用于进一步的体外特性分析。

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