Thymocyte subsets have been well characterized on the basis of CD4 and CD8 antigen expression. Recently, the use of anti-CD3 antibodies has allowed more precise phenotyping of these subsets. The most ...immature T cell precursors are largely CD3-CD4-CD8-, while the most mature are CD3brightCD4+CD8- or CD3brightCD4-CD8+. Moreover, the expression of CD45RA on thymocytes appears to define a progenitor population and may define a continuous lineage of cells. Using a panel of CD45RA antibodies, we have further characterized the CD45RA+ thymocyte population in the murine system. The size of this subset is greatly enhanced in cortisone-treated mice and in sublethally irradiated mice. Moreover, the CD45RA+ population is present early in foetal life and is maintained thereafter. Using three-colour immunofluorescence, we show that (i) while most CD45RA+ cells are present amongst the CD4-CD8- thymocyte subset in the normal thymus, after cortisone treatment or irradiation, all four thymocyte subsets co-express significant amounts of CD45RA. This suggests that not only progenitor cells but also the mature population which can survive such manipulation are CD45RA+; and (ii) a large proportion of CD45RA+ cells are CD3bright and this subset is represented in the thymus at all stages of maturation tested. These data suggest that a proportion of TCR-gamma delta + CD3+ cells in the fetus as well as of TCR-alpha beta+ CD3+ cells in the adult co-express CD45RA.
Mechanisms involved in prothymocyte migration, differentiation and self-commitment were investigated. We used a murine bone marrow fraction isolated on a discontinuous Ficoll gradient and enriched ...10-20 times in CFU-S activity, and studied its fate after intrathymic transfer over a period of 200 days. In order to assess their hemopoietic activity, chimeric thymuses were intravenously transferred to secondary lethally irradiated hosts and both day 8 and day 12 spleen colonies were evaluated. The results show that transfer of a stem cell enriched fraction leads to long-term repopulation of the thymuses and that the input of progenitors is regulated by the size of the intrathymic precursor pool. Furthermore, stem cells can locate within the irradiated thymus and remain in a primitive stage for several months.
This review gives a general overview of the recent advances in the field of fetal hematopoiesis and T-cell development. Although it is now well recognized that early lymphoid progenitors first emerge ...in the fetal liver (FL) where active B lymphopoiesis take places, the identity of early T-cell precursors, as well as the mechanisms of thymus colonization, has long been controversial. Here we discuss the experimental evidence supporting the concept of prothymocyte in both human and mouse species, as well as its major implications regarding lineage relationships among the immune system and general hematopoietic organization.
MRL-lpr/lpr mice have hypertrophied lymph nodes comprising CD4-CD8- T cells. In addition, they contain CD4+CD8- T cells co-expressing the CD45RA marker. The correlation between these two ...subpopulations has been difficult to assess. We analyzed the expression of CD45RA (with the RA3-2C2 antibody) in various thymic and peripheral T cell subsets, using three-color immunofluorescence. We showed that in lpr mice (i) a transient CD4+CD8- thymic subset co-expresses CD45RA during the course of the disease, and (ii) thymic as well as peripheral CD4-CD8- and CD4+CD8- T cells brightly express CD45RA; furthermore (iii) in the lymph nodes, during lymphadenopathy, CD4+CD8-CD45RA+ T cells show a broad range of the CD4 fluorescence intensity, and (iv) the increase in MHC class II expression is restricted to CD45RA-T cells of the thymus and lymph nodes of lpr mice. Taken together, these data suggest that the CD4+CD8-CD45RA+ population might generate the CD4-CD8- tumor cells. In addition, using the bromodeoxyuridine labeling technique, we demonstrate that these cells are not the result of increased proliferation.
The non-obese diabetic (NOD) mouse spontaneously develops a T cell-mediated autoimmune disease, sharing many features with human insulin-dependent diabetes mellitus (IDDM), leading to ...insulin-secreting beta cell destruction. The role of CD4+ T cells has been evidenced at two levels. First, CD4+ T cells from diabetic animals are required to transfer diabetes to non-diabetic recipients in conjunction with CD8+ effector T cells. Second, suppressive CD4+ T cells have been characterized in non-diabetic NOD mice. T cells with different functions can thus share the CD4+ phenotype. Since CD4+ T cells can be divided into at least two subgroups on the basis of CD45 isoform expression, we evaluated the distribution of CD4+ T cells expressing the CD45RA isoform on NOD mouse thymocytes and peripheral T cells. The percentage of CD45RA+ cells was dramatically increased among the most mature CD3bright thymocytes and among CD4+ T cells in lymph nodes of the NOD mouse as compared with control strains. This increase was related to the development of insulitis. Interestingly, the CD45RA isoform was expressed on most CD4+ T cells invading the islets. In vivo treatment with an anti-CD45RA mAb prevented the development of insulitis and spontaneous diabetes in female animals but not the transfer of diabetes by T cells collected from diabetic NOD donors. These results indicate that anti-CD45RA mAb is only effective if given before the full commitment of effector T cells to the destruction of islet beta cells. Thus CD4+CD45RA+ T cells play a key role in early activation steps of anti-islet immunity.
Thymuses of various types of bone-marrow-chimeric mice have been examined by tissue section immunologic staining for the presence and distribution of major histocompatibility complex (MHC) antigens. ...Cortical and medullary thymic epithelial cells continue to express thymus genotype I-A and H-2K/D antigens for at least 6 months posttransplantation. The appearance of bone-marrow-type MHC antigens is limited to low levels of H-2K/D on cortical and medullary lymphocytes, and to dendritic cells in the medulla; the medullary dendritic cells express high levels of donor-type I-A antigens as soon as 3 weeks posttransplantation. The observed patterns support the concept that I-A antigens are synthesized by thymic epithelial cells but are acquired by thymocytes. The findings are of relevance to the understanding of the role of the thymus in the generation of MHC restriction.
Lineage commitment is a fundamental process that is initiated during the early stages of embryogenesis. Ultimately, this leads to the generation of the humoral and cellular arms of the immune system. ...In a recent workshop, researchers from the diverse fields of embryology, haematology and immunology gathered to address the topic of early T-cell differentiation.
Spleen cells from mice neonatally treated with hydrocortisone hemisuccinate (HHC) had an abnormal pattern of cell-mediated immunity when tested two weeks later. Mitogen-induced proliferation was ...reduced. The proliferative responses to allo-antigens were normal but the cytotoxic response thereby induced was impaired. The mechanisms of these anomalies are complex: reduction of cytotoxic precursors as measured by limiting dilution assays; enhancement of suppressor cells which are able to reduce the cytotoxic response of normal spleen cells; reduction of interleukin-2 (Il-2) production. The addition of exogenous Il-2 restores normal proliferative and cytotoxic responses.
