The role of endocytic recycling in autoimmunity.

Department of Medicine, College of Medicine, State University of New York, Upstate Medical University, Syracuse, NY, USA.

Methods in molecular biology (Clifton, N.J.) 2012

Abnormalities in T cell signal transduction underlie pathology in systemic lupus erythematosus. Lupus T cells are more sensitive to stimulation, yet have reduced expression of T cell antigen receptor (TCR) at the surface. The amount of TCR expressed at the surface of a T cell directly determines the ability of a T cell to become activated. The endocytic recycling machinery regulates transport of T cell receptors to the plasma membrane, internalization of surface receptors, and recycling to the cell surface, which determines the ability of a T cell to become activated. Increased recycling of CD3 and CD4 receptors occurs in lupus T cells, and could represent a mechanism by which T cells are sensitized to stimulation. This chapter explains methods used to investigate endocytic recycling of the TCR, CD4, and CD8 co-receptors in peripheral blood lymphocytes, T cells, and in splenocytes from lupus-prone murine models. The assays described will allow the study of surface receptor turnover in live untouched lymphocytes by flow cytometry.