Pedersen Anders Elm, Holmstrøm Kim, Jørgensen Flemming, Jensen Simon S, Gad Monika
Department of International Health, Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen , Copenhagen , Denmark and.
Immunopharmacol Immunotoxicol. 2015 Feb;37(1):63-71. doi: 10.3109/08923973.2014.977449. Epub 2014 Nov 4.
CD4 + CD25+ regulatory T cells (Tregs) are believed to be pivotal in controlling chronic inflammation as well as in opposing the effect of cancer immunotherapy. Therefore, identification of novel drug compounds that interfere with Treg function is of high priority together with research that investigates Treg modulation by current drugs. For such research as well as for novel cell based therapies based on Treg infusions, rapid in vitro assays as well as functional assays based on inhibitory capacity of Tregs are required. Here, we report on such assays using highly pure fluorescence-activated cell sorting (FACS) sorted CD4 + CD25(high)CD127(dim/-)CD45RA+ naïve Treg cells followed by in vitro expansion. We report on the use of these cells in a short-term assay based on Treg mediated inhibition of the early effector T cell activation markers CD69 and CD154. Additionally, we investigate the use of highly pure Tregs in a functional assay based on Treg mediated inhibition of effector T cell proliferation. We report highly reproducible Treg function in assays that test the effect of well-known model compounds such as CpG-A, anti-IL-6R (tocilizumab), anti-TNF-α (adalimumab) or a combination of IL-6 and TNF-α. In conclusion, these assays have the potential for use in pharmacological screening and discovery in relation to drug development in immunology.
CD4 + CD25 + 调节性T细胞（Tregs）被认为在控制慢性炎症以及对抗癌症免疫治疗效果方面起着关键作用。因此，识别干扰Treg功能的新型药物化合物以及研究当前药物对Treg的调节作用是当务之急。对于此类研究以及基于Treg输注的新型细胞疗法，需要快速的体外检测方法以及基于Tregs抑制能力的功能检测方法。在此，我们报告了使用高纯度荧光激活细胞分选（FACS）分选的CD4 + CD25（高）CD127（低/ -）CD45RA + 初始Treg细胞，随后进行体外扩增的此类检测方法。我们报告了在基于Treg介导的早期效应T细胞活化标志物CD69和CD154抑制的短期检测中使用这些细胞的情况。此外，我们研究了在基于Treg介导的效应T细胞增殖抑制的功能检测中使用高纯度Tregs的情况。我们报告了在测试诸如CpG - A、抗IL - 6R（托珠单抗）、抗TNF - α（阿达木单抗）或IL - 6和TNF - α组合等知名模型化合物效果的检测中，Treg功能具有高度可重复性。总之，这些检测方法有潜力用于免疫学药物开发相关的药理筛选和发现。
