School of Chinese Medicine, The University of Hong Kong, Hong Kong, China.
Peking University 3rd Hospital, Peking University Eye Center, Beijing, China.
Methods Mol Biol. 2022;2380:211-224. doi: 10.1007/978-1-0716-1736-6_18.
With the compiling studies on the autoimmune pathogenesis in the developing of Sjögren's syndrome, the functional importance of T follicular helper cells (Tfh) and T follicular regulatory cells (Tfr) was investigated, including our recent findings, among which various techniques for detecting Tfh and Tfr cells in Sjögren's syndrome have been reported. In this chapter, we describe detailed methods for the effective detection of Tfh and Tfr cells in mice with experimental Sjögren's syndrome (ESS), a mouse model with evident salivary hypofunction, increased serum levels of autoantibodies, and histopathological changes in the salivary glands. We provide representative detections of surface markers, cytokines, and transcription factors of Tfh and Tfr cells by flow cytometry and ELISpot assay. Moreover, a detailed protocol for detecting Tfh and Tfr cells in the draining cervical lymph nodes (CLNs) in ESS mice by immunofluorescence microscopy is also described. Together, these techniques of Tfh and Tfr cell detection in ESS mice may facilitate the investigation of autoimmune pathogenesis and enhance the understanding of Tfh and Tfr cell functions in the development of Sjögren's syndrome.
随着对干燥综合征自身免疫发病机制的研究进展，滤泡辅助性 T 细胞（Tfh）和滤泡调节性 T 细胞（Tfr）的功能重要性得到了研究，包括我们最近的发现，其中已经报道了用于检测干燥综合征中 Tfh 和 Tfr 细胞的各种技术。在这一章中，我们描述了在实验性干燥综合征（ESS）小鼠中有效检测 Tfh 和 Tfr 细胞的详细方法，ESS 是一种具有明显唾液功能低下、血清自身抗体水平升高和唾液腺组织病理学改变的小鼠模型。我们通过流式细胞术和 ELISpot 分析提供了 Tfh 和 Tfr 细胞表面标记物、细胞因子和转录因子的代表性检测。此外，还描述了通过免疫荧光显微镜术检测 ESS 小鼠引流颈淋巴结（CLN）中 Tfh 和 Tfr 细胞的详细方案。总之，这些 ESS 小鼠中 Tfh 和 Tfr 细胞检测技术可能有助于研究自身免疫发病机制，并加深对 Tfh 和 Tfr 细胞在干燥综合征发展中的功能的理解。
