Center for Vaccine Development, University of Maryland School of Medicine , Baltimore, MD , USA ; Department of Pediatrics, University of Maryland School of Medicine , Baltimore, MD , USA.
Center for Vaccine Development, University of Maryland School of Medicine , Baltimore, MD , USA ; Department of Medicine, University of Maryland School of Medicine , Baltimore, MD , USA.
Front Immunol. 2014 Jun 19;5:294. doi: 10.3389/fimmu.2014.00294. eCollection 2014.
T cells are the main orchestrators of protective immunity in the stomach; however, limited information on the presence and function of the gastric T subsets is available mainly due to the difficulty in recovering high numbers of viable cells from human gastric biopsies. To overcome this shortcoming we optimized a cell isolation method that yielded high numbers of viable lamina propria mononuclear cells (LPMC) from gastric biopsies. Classic memory T subsets were identified in gastric LPMC and compared to peripheral blood mononuclear cells (PBMC) obtained from children, adults, and the elderly using an optimized 14 color flow cytometry panel. A dominant effector memory T (TEM) phenotype was observed in gastric LPMC CD4(+) and CD8(+) T cells in all age groups. We then evaluated whether these cells represented a population of gastric tissue-resident memory T (TRM) cells by assessing expression of CD103 and CD69. The vast majority of gastric LPMC CD8(+) T cells either co-expressed CD103/CD69 (>70%) or expressed CD103 alone (~20%). Gastric LPMC CD4(+) T cells also either co-expressed CD103/CD69 (>35%) or expressed at least one of these markers. Thus, gastric LPMC CD8(+) and CD4(+) T cells had the characteristics of TRM cells. Gastric CD8(+) and CD4(+) TRM cells produced multiple cytokines (IFN-γ, IL-2, TNF-α, IL-17A, MIP-1β) and up-regulated CD107a upon stimulation. However, marked differences were observed in their cytokine and multi-cytokine profiles when compared to their PBMC TEM counterparts. Furthermore, gastric CD8(+) TRM and CD4(+) TRM cells demonstrated differences in the frequency, susceptibility to activation, and cytokine/multi-cytokine production profiles among the age groups. Most notably, children's gastric TRM cells responded differently to stimuli than gastric TRM cells from adults or the elderly. In conclusion, we demonstrate the presence of gastric TRM, which exhibit diverse functional characteristics in children, adults, and the elderly.
T 细胞是胃中保护性免疫的主要调控者；然而，由于从人类胃活检中回收大量活细胞的困难，关于胃 T 细胞亚群的存在和功能的信息有限。为了克服这一缺点，我们优化了一种细胞分离方法，该方法从胃活检中获得了大量活的固有层单核细胞（LPMC）。使用优化的 14 色流式细胞术面板，在胃 LPMC 中鉴定了经典记忆 T 细胞亚群，并与从儿童、成人和老年人获得的外周血单核细胞（PBMC）进行比较。在所有年龄组中，胃 LPMC CD4（+）和 CD8（+）T 细胞中均观察到主导的效应记忆 T（TEM）表型。然后，我们通过评估 CD103 和 CD69 的表达来评估这些细胞是否代表胃组织驻留记忆 T（TRM）细胞的一个群体。胃 LPMC CD8（+）T 细胞中的绝大多数要么共同表达 CD103/CD69（>70%），要么单独表达 CD103（~20%）。胃 LPMC CD4（+）T 细胞也要么共同表达 CD103/CD69（>35%），要么至少表达其中一种标记物。因此，胃 LPMC CD8（+）和 CD4（+）T 细胞具有 TRM 细胞的特征。胃 CD8（+）和 CD4（+）TRM 细胞在刺激后产生多种细胞因子（IFN-γ、IL-2、TNF-α、IL-17A、MIP-1β）并上调 CD107a。然而，与 PBMC TEM 对应物相比，它们的细胞因子和多细胞因子谱存在明显差异。此外，胃 CD8（+）TRM 和 CD4（+）TRM 细胞在不同年龄组之间在频率、激活易感性和细胞因子/多细胞因子产生谱方面存在差异。值得注意的是，儿童的胃 TRM 细胞对刺激的反应与成人或老年人的胃 TRM 细胞不同。总之，我们证明了胃 TRM 的存在，它们在儿童、成人和老年人中表现出不同的功能特征。
