A Common Epitope Identified by a Monoclonal Antibody, MID 2, Present on All Leucocytes and Associated with a Group of High Molecular Weight Glycopeptides (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.
Clinical & Experimental Immunology
Xenoantisera, designated AT48 and AT72, were developed by immunizing rabbits with human thymus cell membrane and guinea-pigs with a T-cell glycoprotein purified from leukaemic T-cell membrane. Whereas AT48, after appropriate absorption, reacted exclusively with the majority of thymocytes (mainly cortical thymocytes) among normal lymphoid populations, AT72 reacted with virtually all of the thymus and T cells but not with B cells. Thymocytes, which were strongly reactive with AT72, existed in the thymic medulla, but cortical cells were also very weakly reactive with AT72. When cultured T-cell lines, all ofwhich were derived from patients with T-cell-type acute lymphatic leukaemias, were tested for their reactivities with AT48 and AT72 by immunofluorescence, we found that AT48 stained certain T-cell lines, whereas AT72 stained all ofthe T-cell lines tested so far. Immunochemical data showed that AT48 precipitated a 48K molecular weight (mol. wt) glycoprotein from 1251-labelled thymus cell surface glycoproteins, which appeared to be very weakly associated with a 12K mol. wt component. These 48K and 12K mol. wt components precipitated by AT48 showed almost identical isoelectric points (pI) to those of HLA heavy chain and 32-microglobulin respectively. AT72, on the other hand, precipitated a 72K mol. wt glycoprotein from thymus and T cells as well as from leukaemic T cells. A less prominent 65K mol. wt glycoprotein was also precipitated by AT72 from thymus and T cells but not from leukaemic T cells. These two components showed almost identical pl ranging approximately from 4 to 7, and this marked charge heterogeneity observed was reduced by neuraminidase treatment, suggesting that it reflects the heterogeneity in sialylation ofthis molecular species. We concluded from these data that AT48 and AT72 used in this work could detect human homologues ofmouse TL and Ly 1 antigens respectively.
The Journal of Immunology
A glycoprotein complex of 210,000 and 130,000 m.w., found on mitogen or alloantigen-stimulated human T cells and not on other hematopoietic cells, has been defined by a monoclonal antibody (Mab). The components of this complex are a subset of a larger family of proteins (210,000, 165,000 and 130,000 m.w.) defined by a second Mab. In a panel of hematopoietic cell lines and cell types, only activated T cells (including the cell line HUT-102) express the 210,000/130,000 complex and these cells also express the IL 2 receptor, a characteristic marker for activated T cells. The 210,000/130,000 m.w. complex (reactive with the Mab TS2/7) is present on all long-term activated T cells, including both the OKT4 and OKT8 subsets. The 210,000 m.w. subunit is expressed only on activated T cells. Other lymphoid cells express either the 130,000 m.w. subunit alone (unactivated lymphocytes, thymocytes, HUT-78) or the 130,000 subunit together with a 165,000 subunit (MOLT-4, HSB, and other leukemic T ce...
Antigenic Pattern of Human Brain Glycoproteins as Described by Monoclonal Antibodies
Journal of Neurochemistry, 1987
A battery of monoclonal antibodies was raised against a preparation of lentil lectin-binding membrane glycoproteins from human brain. Out of 26 established hybridomas, nine produced antibodies against the human Thy-1 antigen. For the remaining 17 lines, reactivity with at least six other antigens could be identified after immunoprecipitation and immunoblotting. Several of the antigens were dior trimeric, mainly in the molecular weight range of 60-120 kDa. Two of the antibodies were reactive with highmolecular-weight aggregates and four targets for the anti
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.
Biochimica et biophysica acta, 1975
A protein immunologically identical to a glycoprotein antigen from fibroblast plasma membrane (SF antigen complex) is present in chicken serum. This antigen disappears from cells transformed with tumor viruses. The antigen solubilized from fibroblasts using urea and detergents, and the serum component both gave a molecular weight of about 2-10(5) as estimated by gel filtration on Sepharose 4B. Ultracentrifugation in 5-20% sucrose gradients gave a value of 7-8 S for the antigen from both sources. Isolation of antigen from serum and fibroblasts extracts was achieved using immunochemical techniques. Analysis of the polypeptide chains of the cellular antigen by electrophoresis in the presence of sodium dodecylsulphate, disclosed three major components at molecular weights 210 000 and 145 000 and 45 000. The two high molecular weight bands were identified in electrophoretograms of whole cell extracts by absorption to and subsequent elution from immunoadsorbents. The 45 000-molecular weig...
Materials and methods for analysis of Glycoprotein components
The plasma glycoproteins were precipitated with alcohol. To 0.1 ml of plasma added 2.0 ml of alcohol and centrifuged. The supernatant was decanted. The precipitate was hydrolyzed with acid to liberate protein bound hexose, hexosamine and sialic acid.
III. A~tr~o AcID COMPOSITION OF FOUR ANTIBODIES FROII ONE INDIVIDUAL
The amino acid composition of pooled human 7-globulin was studied comprehensively by Brand and coworkers (1). More recent studies of 6.5S human globulins, now called 7G-globulin (2), using automatic zmlno acid analytical techniques (3), have been reported for example, by Hsaio and Putnam (4), and by Crumpton and Wilkinson (5). In contrast to the situation with amino acid analyses of specific antibodies from the rabbit (5-10), the residue composition of human antibody 3,-globulins to well defined antigens, especially from individual subjects, has yet to be investigated. This paper is concerned with a study of the amino acid composition of four purified 7G-antibodies, namely antilevan, antidextran, antiteichoic acid, and antiblood group substance A, from the serum of one individual. These four antibody preparations were previously examined for their allotypic speciiicities (11) and for the electrophoretic patterns of their H and L chains in starch gel (12).