Developmental regulation of lymphocyte-specific protein 1 (LSP1) expression in thymus during human T-cell maturation

The lymphocyte specific protein 1 (LSP1) phosphoprotein is an F-actin binding molecule restricted to cells of hematopoietic origin in mice and humans. LSP1 is localized to the internal surface of the plasma membrane, the cytoplasm, and NP-40-insoluble actin filaments and;is thought to mediate cytoskeleton-driven responses in activated leukocytes that involve receptor capping, cell-cell interactions and cell motility. Here, we generated two monoclonal antibodies (MAbs), 5E3 and 14G8, that are specific for human LSP1 to define the expression of LSP1 throughout human T-cell development. Both MAbs reacted with a 52-kDa protein in BW5147 cells transfected with human LSP1 cDNA in pcDNA3, but not in cells transfected with cDNA in an antisense orientation, indicating the specificity of 5E3 and 14G8 for human LSP1. In developing T cells, LSP1 was expressed on human fetal liver CD7+ NK and T-cell precursors, the CD7+, CD3-, CD4-, CD8- human stem cell line DU-528, and on CD4-, CD8- double-negative (DN) thymocytes. Immunohistochemistry and three-color flow cytometry analysis of fetal or postnatal thymocytes revealed that LSP1 was increasingly expressed during intrathymic human T-cell maturation. While immature CD4+CD8+ double-positive (DP) thymocytes expressed low to undetectable levels of LSP1, mature CD4+CD8- and CD4-CD8+ single-positive (SP) thymocytes expressed high levels of LSP1. Thus, LSP1 is developmentally regulated during T-cell maturation within the human thymus and may play a functional role in the motility of DN and SP thymocytes.