Methods in Molecular Biology Volume 900, 2012, pp 91-107

The Role of Endocytic Recycling in Autoimmunity

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Abstract

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.