Detection of T Follicular Helper Cells and T Follicular Regulatory Cells in Experimental Sjögren’s Syndrome

Abstract

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.