Phenotypic and functional properties of murine thymocytes. I. Precursors of cytolytic T lymphocytes and interleukin 2-producing cells are all contained within a subpopulation of "mature" thymocytes as analyzed by monoclonal antibodies and flow microfluorometry
Open Access
- 1 February 1982
- journal article
- research article
- Published by Rockefeller University Press in The Journal of Experimental Medicine
- Vol. 155 (2), 358-379
- https://doi.org/10.1084/jem.155.2.358
Abstract
The correlation between surface phenotype and function in subpopulations of murine thymocytes was investigated using flow microfluorometry (FMF). C57BL/6 thymocytes stained with monoclonal antibodies directed against Lyt-2, H-2Kb, and Thy-1.2 and passed on a fluorescence-activated cell sorter (FACS II flow cytometer) could be resolved into at least 4 distinct subpopulations on the basis of fluorescence and forward light scatter measurements: (a) medium-sized Lyt-2+ cells that stained strongly with H-2Kb and weakly with Thy-1.2 (5% of total cells); (b) medium-sized Lyt-2- cells with other properties as in (a) (10%); (c) small Lyt-2+ cells that stained weakly with H-2Kb and strongly with Thy-1.2 (60%); and (d) large Lyt-2+ cells that stained weakly with H-2Kb and very strongly with Thy-1.2 (23%). Cortisone-resistant (CR) thymocytes were found to correspond phenotypically to populations (a) and (b). The distribution of cytolytic T lymphocyte precursors (CTL-P) directed against H-2d alloantigens in subpopulations of C57BL/6 thymocytes that were sorted according to the phenotypic criteria described above was then investigated. CTL-P in sorted and control populations were quantitated by limiting dilution analysis of mixed leukocyte microcultures established in an excess of interleukin 2 (IL-2). All thymus CTL-P apparently could be quantitatively recovered in a subpopulation of cells that was CR, medium-sized, Lyt-2+, H-2Kb+ and weakly stained with Thy-1.2. In parallel studies, the production of IL-2 by subpopulations of C57BL/6 thymocytes was quantitatively assessed using a sensitive microassay system. Graded numbers of sorted or control thymocytes were stimulated with irradiated T cell-depleted allogeneic cells and assayed for their ability to support the growth of an IL-2-dependent cytolytic T lymphocyte clone. IL-2 production was found to reside entirely in a subpopulation of CR, medium-sized, Lyt-2- thymocytes. Further phenotypic analysis of this subpopulation of cells indicated that it was homogeneously H-2Kb+ and weakly staining with Thy-1.2. Thus, subpopulation of thymocytes with a mature phenotype (i.e., Cr, medium-sized, H-2Kb+ and weakly staining with Thy-1.2) accounts for all the functional activity in the thymus. Reasons for the apparent discrepancy between these results and other recent studies are discussed.Keywords
This publication has 39 references indexed in Scilit:
- Functional subclasses of T lymphocytes bearing different Ly antigens. II. Cooperation between subclasses of Ly+ cells in the generation of killer activity.The Journal of Experimental Medicine, 1975
- Thymus cell maturation: II. Differentiation of three “mature” subclasses in VivoCellular Immunology, 1975
- Subpopulations of T-lymphocytes. Physical separation, functional specialisation and differentiation pathways of sub-sets of thymocytes and thymus-dependent peripheral lymphocytes.1975
- GENERATION OF CYTOTOXIC T LYMPHOCYTES IN VITROThe Journal of Experimental Medicine, 1974
- Allograft immunity in the mouse. II. Physical studies of the development of cytotoxic effector cells from their immediate progenitors.1973
- Cell-mediated immune response in vitro: II. The role of thymus and thymus-derived lymphocytesCellular Immunology, 1972
- SYNERGY AMONG LYMPHOID CELLS MEDIATING THE GRAFT-VERSUS-HOST RESPONSEThe Journal of Experimental Medicine, 1972
- Characteristics of the immunocompetent cells in the mouse thymus: Cell population changes during cortisone-induced atrophy and subsequent regenerationCellular Immunology, 1970
- Current concepts of the immunological function of the thymus.Physiological Reviews, 1967
- LOCALIZATION OF MOUSE ISOANTIGENS ON CELL SURFACE AS REVEALED BY IMMUNOFLUORESCENCE1967