ABSTRACT
Ectopic expression of insulin in thymus has been suggested to be involved in tolerance induction against pancreatic beta cells and in type 1 diabetes (T1D) pathogenesis. However, it is not known whether thymic insulin expression would also influence thymocyte maturation and differentiation. To address these questions, we have used mice that are insulin deficient. Early fetal thymi were cultured in fetal thymic organ cultures (FTOCs) and the development of thymocytes was studied by flow cytometry. The results revealed no significant difference in thymocyte maturation in the absence of thymic insulin. Taken together, these data do not support a role for thymic insulin in thymocyte differentiation and growth.
Subject(s)
Insulin/deficiency , T-Lymphocytes/cytology , T-Lymphocytes/metabolism , Thymus Gland/cytology , Thymus Gland/embryology , Animals , Cell Differentiation , Cells, Cultured , Female , Flow Cytometry , Insulin/genetics , Insulin/metabolism , Mice , Mice, Mutant Strains , Organ Culture Techniques , T-Lymphocytes/immunology , Thymus Gland/immunology , Time FactorsABSTRACT
We have taken advantage of the Cre/lox system to generate a mouse model with inducible deficiency of transforming growth factor beta receptor II (TbetaRII). Using this approach, transforming growth factor beta (TGF-beta) signaling deficiency can be restricted to the hematopoietic system by bone marrow transplantation. Mice that received transplants with TbetaRII-/- bone marrow develop a lethal inflammatory disorder closely resembling that of TGF-beta1-null mice. Previous in vitro studies have suggested multiple roles for TGF-beta in T-cell development, including proliferation, apoptosis, and differentiation. We used our transplantation model to ask whether T-cell development is normal in the absence of TGF-beta signaling. The findings show for the first time in vivo and in fetal thymus organ culture (FTOC) that TGF-beta is not required for thymocytes to differentiate along the entire pathway of thymic T-cell development, as defined by the expression patterns of CD4, CD8, CD25, and CD44. In contrast to previous investigations, no increase of thymocyte apoptosis was observed. However, TbetaRII-deficient CD8+ thymocytes displayed a 2-fold increase in proliferation rate, as determined by bromodeoxyuridine (BrdU) incorporation in vivo. These results reinforce the importance of TGF-beta as an immune regulator critical for T-cell function.
