Abstract
Abstract Retinoic acid-related Orphan Receptor gamma t (RORγt) is a DNA-binding transcription factor that is selectively expressed by lymphoid cells including CD4+CD8+ thymocytes, CD4+ Th17 and CD8+ Tc17 cells, NKT cells and lymphoid tissue inducer (LTi) cells. RORγt plays essential roles in thymopoiesis, T cell homeostasis, effector T cell differentiation and the development of secondary lymphoid tissues, eg, lymph nodes and Peyer’s patches. A new mouse mAb, Q31-378, that recognizes mouse RORγt was developed to provide a sensitive probe for analyzing the cellular and molecular mechanisms that underlie the development and differentiation of T cells and lymphoid tissues. With a newly developed transcription factor buffer, Q31-378 was successfully used in multicolor flow cytometric analyses to analyze RORγt expression in thymocyte subsets defined by CD4 and CD8 expression. Among all thymocytes, CD4+CD8+ cells comprised the main cell subset that expressed RORγt. Analyses of splenocytes cultured under Th17-inducing conditions revealed that most activated CD4+IL-17A+IFN-γ± T cells expressed RORγt when compared with other T cell subsets. These new reagents, along with additional fluorescent antibodies to other cytokines, transcription factors and phenotypic markers are being applied to analyze RORγt expression patterns in the development and differentiation of T cells and LTi cells.
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