Kirchhoff Dennis, Frentsch Marco, Leclerk Patrick, Bumann Dirk, Rausch Sebastian, Hartmann Susanne, Thiel Andreas, Scheffold Alexander
Immunomodulation Group, Deutsches Rheuma-Forschungszentrum Berlin, Berlin, Germany.
Eur J Immunol. 2007 Sep;37(9):2370-7. doi: 10.1002/eji.200737322.
T helper (Th) cells are central regulators of adaptive immune responses. However, the detection of the small number of Th cells specific for a particular antigen or pathogen is still a major challenge. CD154 was recently introduced as a marker for antigen-specific Th cells. To date, this technology was not applicable for mice - arguably the most important immunological model system. CD154 is difficult to detect due to its rapid removal from the cell surface upon binding to CD40 during antigen-specific activation by APC. We present an efficient strategy to block the degradation of murine CD154 by combined use of antibodies against CD40 and CD154. This strategy makes CD154 easily accessible for surface staining, which allows isolation and expansion of rare antigen specific T cells. Importantly, CD154 identified all specific T cells in strongly Th1- or Th2-polarized immune responses against pathogens like Salmonella typhimurium and Heligmosomoides polygyrus, independent of their potential to produce cytokines. We demonstrate that CD154 can in fact be used as a reliable marker for antigen-specific CD4 T cells in mice, offering a unique option to analyze, isolate and rapidly expand the entire pool of Th-cells generated during a physiological T cell response in vivo.
辅助性T（Th）细胞是适应性免疫反应的核心调节因子。然而，检测针对特定抗原或病原体的少量Th细胞仍然是一项重大挑战。CD154最近被引入作为抗原特异性Th细胞的标志物。迄今为止，这项技术不适用于小鼠——可以说是最重要的免疫学模型系统。由于在抗原呈递细胞（APC）介导的抗原特异性激活过程中，CD154与CD40结合后会迅速从细胞表面清除，因此难以检测到它。我们提出了一种有效的策略，通过联合使用抗CD40和抗CD154抗体来阻断小鼠CD154的降解。该策略使CD154易于进行表面染色，从而能够分离和扩增罕见的抗原特异性T细胞。重要的是，在针对鼠伤寒沙门氏菌和多房棘球绦虫等病原体的强烈Th1或Th2极化免疫反应中，CD154能够识别所有特异性T细胞，而与它们产生细胞因子的潜力无关。我们证明，CD154实际上可以用作小鼠中抗原特异性CD4 T细胞的可靠标志物，为分析、分离和快速扩增体内生理性T细胞反应过程中产生的整个Th细胞库提供了一个独特的选择。
