notch signalling inhibits cd4 expression during initiation and differentiation of human t cell lineagenotch信号抑制cd4的表达在起始和人类t细胞谱系分化.pdfVIP

Notch Signalling Inhibits CD4 Expression during Initiation and Differentiation of Human T Cell Lineage 1 1 1,2 2 Stephen M. Carlin , Melissa L. M. Khoo , David D. Ma , John J. Moore * 1 Blood Stem Cells and Cancer Research, St Vincent’s Centre for Applied Medical Research, Sydney, New South Wales, Australia, 2 Haematology Department, St Vincent’s Hospital, Sydney, New South Wales, Australia Abstract The Delta/Notch signal transduction pathway is central to T cell differentiation from haemopoietic stem cells (HSCs). Although T cell development is well characterized using expression of cell surface markers, the detailed mechanisms driving differentiation have not been established. This issue becomes central with observations that adult HSCs exhibit poor differentiation towards the T cell lineage relative to neonatal or embryonic precursors. This study investigates the contribution of Notch signalling and stromal support cells to differentiation of adult and Cord Blood (CB) human HSCs, using the Notch signalling OP9Delta co-culture system. Co-cultured cells were assayed at weekly intervals during development for phenotype markers using flow cytometry. Cells were also assayed for mRNA expression at critical developmental stages. Expression of the central thymocyte marker CD4 was initiated independently of Notch signalling, while cells grown with Notch signalling had reduced expression of CD4 mRNA and protein. Interruption of Notch signalling in partially differentiated cells increased CD4 mRNA and protein expression, and promoted differentiation to CD4+ CD8+ T cells. We identified a set of ge