Yirsaw Alehegne W, Gillespie Alexandria, Britton Emily, Doerle Alyssa, Johnson Lisa, Marston Susan, Telfer Janice, Baldwin Cynthia L
Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA, 01003, USA.
Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA, 01003, USA; Program in Molecular and Cellular Biology, University of Massachusetts, Amherst, MA, 01003, USA.
Dev Comp Immunol. 2021 May;118:103984. doi: 10.1016/j.dci.2020.103984. Epub 2020 Dec 19.
The major functions of γδ T cells in mammals overlap with those of αβ T cells but differ in that γδ T cells are rapid responders and see different types of antigens. While γδ T cells have been shown to be a major population of circulating lymphocytes in artiodactyl species such as cattle, sheep, and pigs, less is known about these cells in goats, an important agricultural species. We have recently shown that WC1, a γδ T cell-specific family of hybrid pattern recognition receptors/co-receptors, is a multigenic family in goats expanded beyond what occurs in cattle. This study was conducted to address some of the limitations of previous studies in determining the proportions of γδ T cells, WC1 γδ T cells as well as the WC1.1 and WC1.2 subpopulations in blood and to evaluate their responses to various pathogens. Previously, the proportion of caprine γδ T cells was determined using a monoclonal antibody (mAb) 86D that we show here does not react with all γδ T cells thereby underestimating their contribution to the lymphocyte population. Using a mAb reactive with the TCRδ constant region we found the proportion of γδ T cells in blood was not significantly less than that of either CD4 or CD8 T cells and did not decrease with age after 6 months. γδ T cells that expressed WC1 ranged from ~20 to 85% of the total γδ T cells. Less than half of those were classified as WC1.1 or WC1.2 by mAb staining thus indicating a third major WC1 population. We found that naïve γδ T cells proliferated in cultures of PBMC stimulated with antigens of Leptospira or Mycobacterium avium paratuberculosis (MAP) more than they did in control medium cultures or in those stimulated with M. bovis BCG antigens and that the responding γδ T cells included both WC1 and WC1 cells. In ex vivo PMA/ionomycin-stimulated cultures of WC1 γδ T cells but not WC1 cells produced both IL-17 and IFNγ. In longterm cultures with Leptospira or MAP both WC1 and WC1 cells proliferated but only WC1 γδ T cells produced IL-17. In conclusion, goats have a substantial number of WC1 and WC1 γδ T cells in PBMC that do not decrease with animal age after 6 months; both populations respond to bacterial antigens as naïve cells but in these cultures only the WC1 γδ cells produc IL-17 and IFNγ .
哺乳动物中γδ T细胞的主要功能与αβ T细胞的功能重叠,但不同之处在于γδ T细胞反应迅速,识别的抗原类型也不同。虽然γδ T细胞已被证明是牛、羊和猪等偶蹄目动物循环淋巴细胞的主要组成部分,但对于山羊这种重要的农业物种中的这些细胞,我们了解得较少。我们最近发现,WC1是γδ T细胞特异性的杂合模式识别受体/共受体家族,在山羊中是一个多基因家族,其扩增程度超过了牛。本研究旨在解决以往研究在确定血液中γδ T细胞、WC1 γδ T细胞以及WC1.1和WC1.2亚群比例方面的一些局限性,并评估它们对各种病原体的反应。以前,山羊γδ T细胞的比例是使用单克隆抗体(mAb)86D来确定的,我们在此表明该抗体并非与所有γδ T细胞都发生反应,因此低估了它们在淋巴细胞群体中的贡献。使用与TCRδ恒定区反应的单克隆抗体,我们发现血液中γδ T细胞的比例并不显著低于CD4或CD8 T细胞,并且在6个月后不会随年龄增长而减少。表达WC1的γδ T细胞占总γδ T细胞的比例约为20%至85%。通过单克隆抗体染色,其中不到一半被归类为WC1.1或WC1.2,这表明存在第三个主要的WC1群体。我们发现,用钩端螺旋体或副结核分枝杆菌(MAP)抗原刺激外周血单核细胞(PBMC)培养物时,幼稚γδ T细胞的增殖比在对照培养基培养物或用牛分枝杆菌卡介苗抗原刺激的培养物中更明显,并且反应性γδ T细胞包括WC1和WC1细胞。在体外经佛波酯/离子霉素刺激的WC1 γδ T细胞培养物中,但不是WC1细胞培养物中,会产生IL-17和IFNγ。在与钩端螺旋体或MAP的长期培养中,WC1和WC1细胞都会增殖,但只有WC1 γδ T细胞会产生IL-17。总之,山羊外周血单核细胞中有大量的WC1和WC1 γδ T细胞,6个月后不会随动物年龄增长而减少;这两个群体作为幼稚细胞对细菌抗原都有反应,但在这些培养物中只有WC1 γδ细胞会产生IL-17和IFNγ 。