Studies of a human T lymphocyte antigen recognized by a monoclonal antibody (original) (raw)
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Clinical & Experimental Immunology
Human thymus and T cell antigens were identified by using four distinct monoclonal antibodies (MoAb), designated 2D5, 5B3, 7A5 and 9D4. 2D5 antibody reacted with most human thymocytes and a few peripheral lymphocytes as well as with a subpopulation (20%) of bone marrow cells, and precipitated a 45K molecular weight (mol. wt.) component from '251-labelled thymus cell lysate. 7A5 antibody also reacted with the majority (80%) of thymocytes but neither with peripheral lymphocytes nor with bone marrow cells. The antigen detected by 7A5 was a glycoprotein consisting of 48K and 12K mol. wt. components, which were non-covalently associated with each other. 5B3 reacted with virtually all of human thymus and T cells but not with the majority of B cells and bone marrow cells. This reagent precipitated a 72K mol. wt. glycoprotein from thymus and T cells. An additional 65K mol. wt. glycoprotein was precipitated by 5B3 together with the 72K mol. wt. component, but with poor reproducibility. 9D4 antibody, on the other hand, reacted with a 200K mol. wt. component from thymus and T cells as well as 220K and 210K components from the non-T cell fraction of tonsil lymphocytes. Whereas antigens detected by 2D5 and 7A5 appeared to be highly expressed on cortical thymocytes, the antigen defined by 5B3 occurred much more abundantly on medullary thymocytes and peripheral T cells than on cortical thymocytes. Based on the data described above, it is suggested that 7A5, 5B3 and 9D4 MoAb recognize human homologues of mouse TL, Ly-1 and Ly-5 antigens, respectively, whereas 2D5 antibody seems to resemble OKT10, as described by others.
Identification of a novel subset of human T lymphocytes defined by monoclonal antibodies
Immunology
A novel subset of human blood lymphocytes was isolated by means of labelling with monoclonal antibodies and fluorescence-activated cell sorting. In normal individuals, the new subset accounts for about 2% of the blood T lymphocytes. The cells of this subset bind monoclonal antibodies specific for T lymphocytes in general [e.g. OKT3, Hu-Lyt 3 (9 6) and Leu-l] and they also form E rosettes. However, no binding is seen with monoclonal antibodies to T-lymphocyte subsets (OKT4, OKT8, Leu-2A and Leu-3A). Moreover, the lymphocytes of this new subset express neither Ia antigens nor membrane immunoglobulins. They do not bind OKM 1, an antibody against cells of the myelomonocytic lineage that also reacts with natural killer cells, nor do they bind OKT6 or OKT10, 313 specific for thymocyte antigens. The cells have a high specific gravity, a thymocyte-like pattern of lactate dehydrogenase isoenzymes and do not contain terminal deoxynucleotidyl transferase. Although these lymphocytes are viable, also after culture in vitro, and can be stored in liquid nitrogen, they are inert in all functional systems tested: they neither proliferate upon stimulation with mitogens or allogeneic cells, nor do they display suppressor or natural killer cell activity. A patient who was successfully reconstituted by bone marrow transplantation for severe combined immunodeficiency, was found to contain an abnormally high (25%-30%) fraction of these OKT3 positive, OKT4 and OKT8 negative cells among his circulating T lymphocytes.
Isolation of T lymphocyte subsets from peripheral blood using monoclonal antilymphocyte antibodies
Annals of clinical and laboratory science
Monoclonal antisera against helper (serum OKT4) and suppressor (serum OKT8) T cells were employed in a complement-dependent lymphocytotoxicity reaction to isolate helper and suppressor T cells from peripheral blood mononuclear cells. When prepared by this method, helper and suppressor T cells had a purity of 67.8 percent and 78.8 percent, respectively. Approximately 56 percent of viable suppressor and 33 percent of viable helper T cells were lost during the procedure.
Journal of Immunological Methods, 1980
An anti-human T-lymphocyte antiserum has been further studied for specificity for T-cell subpopulations and application in tissue sections. Using a complement-dependent microcytotoxicity assay about 60% of normal peripheral blood T-cells were found to be sensitive to lysis, while 40% were resistant. When the T-cell suspensions were depleted of TG-cells using EA (Ripley) rosette sedimentation, the remaining cells demonstrated an increased percentage of lysis-sensitive cells, while the T-cells enriched in EA-RFC demonstrated a decreased percentage of cells sensitive to lysis, indicating that the antiserum was primarily directed against the non TG-Cetls , i.e. probably the T M helper cell population. This was further supported by functional studies. In order to quantitate T-cells, cytocentrifuge preparations were made from cell suspensions with known T-cell percentages and the T-cells determined with both the immune adherence technique using AET-treated sheep erythrocytes, as well as the indirect immunofluorescence technique using anti-T antiserum. The results of the two methods correlated well, suggesting that both methods can be used to determine T-cells in situ in infiltrate-like clusters of cells.
A novel surface antigen expressed by mature and activated rabbit T-lymphocytes
Cellular Immunology, 1990
The monoclonal antibody (Mab) 93C6 was used to identify a surface molecule expressed by mature and activated rabbit T-lymphocytes. This antigen was virtually absent in cells of the thymus (5% positive cells). All 93C6-positive cells expressed the rabbit panT marker L I l-135 and proliferated in response to concanavalin A (Con A). When lymphocytes were activated in the presence of Con A, the expression of the 93C6 molecule was quantitatively increased. Con A stimulation alone did not markedly enhance expression of this antigen by thymocytes, but stimulation with both Con A and interleukin-2 (IL-2) resulted in a population of thymocytes that expressed the antigen in a manner quantitatively similar to that of mature T-cells in other tkSUeS.
Monoclonal antibodies against T-cell antigens studied by immunohistochemistry
Blut, 1982
Fifteen monoclonal antibodies against different T-cell antigens were studied by immunohistochemistry in thymus, fetal thymus, fetal liver, palatine tonsils, and a few T-cell lymphomas. OKT 9 was identified as reacting with hemopoietic stem or precursor cells in fetal liver as well as with early B-determined lymphocytes in tonsillar germinal centres. OKT 10 labelled lymphocytes in thymus and surprisingly also the cytoplasm of some tonsillar cells with plasma-cell like appearance. OKT 6 and MAS 036 b reacted only with thymic cells. OKT 4, OKT 5, OKT 8, 8-11, labelled thymic cells-and portions of interfollicular cells in tonsils. OKT 3, NEI 016, NEI 015, and T 28 stained a majority of thymic cells and of tonsillar interfollicular lymphocytes. IFH-M 203, NE1012 and 4-11 were positive with the majority of T-lymphocytes in tonsils but labelled only a few thymic cells.
The Human Cytolytic T Lymphocyte Response to Transplantation Antigens
Pediatric Research, 1985
Cytolytic T lymphocytes are important ef-using panels of EBV lines. For example, shows that a fectors in the immune response to allografts, viral infeccytolytic T lymphocyte clone that was stimulated by the cell JY tions, and some tumors. Cytolytic T lymphocyte lines and (HLA-A2,2; B7,7,DR4,6) causes significant cytolysis of JY and clones have been generated and used to define functionally Priess but not of a variety of other cell types. An extended panel relevant effector and target cell surface molecules. The showed that this CTL line was specific for HLA-A2. Similar major target antigens of the human allogeneic response are target panels have demonstrated other CTL clones specific for the major histocompatibility complex antigens. Function-HLA-A2 (4, 5), -B7 (4, 5), -DR6 (4, 6), and DC1 . Other ally relevant effector antigens include LFA-1, LFA-2, authors have also reported a variety of CTL that are specific for LFA-3,OKT4, OKTS, OKT3, and Ti, the T cell receptor. HLA class I and class I1 antigens (reviewed in Ref. 8).