Pathogen-Reactive T Helper Cell Analysis in the Pig
Pathogen-Reactive T Helper Cell Analysis in the Pig
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DOI：
10.3389/fimmu.2017.00565
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发表时间：
2017-05-17
影响因子：
7.3
通讯作者：
Hartmann, Susanne
中科院分区：
文献类型：
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作者：
Ebner, Friederike;Schwiertz, Patrycja;Hartmann, Susanne
在与人类相关的大型动物模型（如猪）中研究宿主-病原体相互作用的兴趣越来越大。尽管猪T细胞研究的免疫学试剂的开发取得了进展，但仍迫切需要直接评估病原体特异性T细胞-一种极其罕见的细胞群体，但在协调宿主对给定病原体的免疫反应方面至关重要。在这里，我们确定了从人类和小鼠研究中已知的活化标志物CD 154（CD 40 L）也可以识别猪抗原反应性CD 4（+）T淋巴细胞。CD 154表达在抗原接触后早期上调，并且CD 4（+）CD 154（+）抗原反应性T细胞共表达细胞因子。抗原诱导的扩增和自体再刺激使得能够对CD 154调节进行时间和剂量分辨分析，并显著提高抗原应答细胞表型分析的分辨率。CD 154表达鉴定了响应于葡萄球菌肠毒素B超抗原刺激的T细胞，以及响应于真菌白色念珠菌的T细胞和对高度流行的肠道寄生虫（猪蛔虫线虫）特异的T细胞。用猪链球菌单一重组细菌抗原免疫猪后，进一步检测到抗原反应性T细胞。因此，我们的研究提供了新的方法来研究抗原特异性T淋巴细胞在猪和它们的贡献宿主-病原体相互作用。
There is growing interest in studying host-pathogen interactions in human-relevant large animal models such as the pig. Despite the progress in developing immunological reagents for porcine T cell research, there is an urgent need to directly assess pathogen-specific T cells-an extremely rare population of cells, but of upmost importance in orchestrating the host immune response to a given pathogen. Here, we established that the activation marker CD154 (CD40L), known from human and mouse studies, identifies also porcine antigen-reactive CD4(+) T lymphocytes. CD154 expression was upregulated early after antigen encounter and CD4(+)CD154(+) antigen-reactive T cells coexpressed cytokines. Antigen-induced expansion and autologous restimulation enabled a time-and dose-resolved analysis of CD154 regulation and a significantly increased resolution in phenotypic profiling of antigen-responsive cells. CD154 expression identified T cells responding to staphylococcal Enterotoxin B superantigen stimulation as well as T cells responding to the fungus Candida albicans and T cells specific for a highly prevalent intestinal parasite, the nematode Ascaris suum during acute and trickle infection. Antigen-reactive T cells were further detected after immunization of pigs with a single recombinant bacterial antigen of Streptococcus suis only. Thus, our study offers new ways to study antigen-specific T lymphocytes in the pig and their contribution to host-pathogen interactions.