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  • 1
    Online Resource
    Online Resource
    Rockefeller University Press ; 1974
    In:  The Journal of Experimental Medicine Vol. 140, No. 2 ( 1974-08-01), p. 452-469
    In: The Journal of Experimental Medicine, Rockefeller University Press, Vol. 140, No. 2 ( 1974-08-01), p. 452-469
    Abstract: To determine whether or not B lymphocytes are committed to the synthesis of a single immunoglobulin heavy chain isotype during their differentiation into plasma cells, rabbit lymph node and Peyer's patch cells were separated into populations with and without membrane IgM, using a fluorescence-activated cell sorter (FACS). The potential of the µ-bearing (µ+) and non-µ-bearing (µ-) cells to give rise to plasma cells both in vivo after transfer into irradiated recipients and in vitro in the presence of pokeweed mitogen was assessed by immunofluorescence techniques, and the relative proportions of the cytoplasmic Ig-stained cells (CSC) synthesizing each class of heavy chains were determined. Most of the CSC arising in vitro from µ-bearing lymph node and Peyer's patch cells contained IgM; all IgM CSC appeared to be derived from µ+ cells. Peyer's patch lymphocytes, however, did not generate IgM CSC after cell transfer and thus may be functionally different from lymph node µ+ cells. It was found also that nearly all of the many IgA CSC generated by Peyer's patch lymphocytes either in culture or after transfer were derived from µ- cells. Further fractionation of these µ- cells with the FACS after they had been membrane stained with anti-b locus allotype reagents revealed that the precursors of IgA CSC belong to a minor population of cells which do have b locus light chain determinants on their membranes, although they do not have detectable µ-chains. These cells are not found in lymph nodes. Although the majority of Peyer's patch and lymph node cells were found to be precommitted to the synthesis of a single heavy chain isotype, a small proportion of cells may not be similarly restricted. Some of the CSC with membrane IgM were found to contain cytoplasmic IgA or IgG. In addition, µ+ populations did give rise to low numbers of IgA and IgG CSC. The implications of these results, obtained under experimental conditions, on the normal differentiation of B lymphocytes in situ are discussed.
    Type of Medium: Online Resource
    ISSN: 1540-9538 , 0022-1007
    RVK:
    Language: English
    Publisher: Rockefeller University Press
    Publication Date: 1974
    detail.hit.zdb_id: 1477240-1
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  • 2
    Online Resource
    Online Resource
    The American Association of Immunologists ; 1973
    In:  The Journal of Immunology Vol. 111, No. 5 ( 1973-11-01), p. 1334-1348
    In: The Journal of Immunology, The American Association of Immunologists, Vol. 111, No. 5 ( 1973-11-01), p. 1334-1348
    Abstract: Lymphocytes from peripheral blood and Peyer's patches of b5b9 and b4b5 rabbits were membrane stained with rhodamine- and fluorescein-labeled anti-allotype reagents. Up to 63% of peripheral blood lymphocytes and 15% of Peyer's patch lymphocytes were found to double stain for both allotypes on their membranes. With a fluorescence-activated electronic cell sorter, cell populations were fractionated according to membrane allotype. A double pass separation procedure after staining yielded four distinct cell populations: one each of cells single-stained for one of the two allotypes, one containing cells double-stained for both allotypes, and one of unstained cells. Properties of the cells bearing two allotypes were examined. Cells put into culture after they were stripped of their membrane immunoglobulin (Ig) with Pronase regenerated their membrane Ig; however, the proportion of cells bearing both allotypes was greatly reduced. This suggested that the lymphocytes were restricted to the synthesis of membrane Ig molecules of a single allotype at a given time. It was found that a high proportion of rabbit lymphocytes could bind exogenous Ig from serum in vitro and probably also in vivo, explaining the presence of molecules of both allotypes on individual cells.
    Type of Medium: Online Resource
    ISSN: 0022-1767 , 1550-6606
    RVK:
    RVK:
    Language: English
    Publisher: The American Association of Immunologists
    Publication Date: 1973
    detail.hit.zdb_id: 1475085-5
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  • 3
    Online Resource
    Online Resource
    Rockefeller University Press ; 1974
    In:  The Journal of Experimental Medicine Vol. 140, No. 4 ( 1974-10-01), p. 966-976
    In: The Journal of Experimental Medicine, Rockefeller University Press, Vol. 140, No. 4 ( 1974-10-01), p. 966-976
    Abstract: Fluorescent antibody staining with antibodies to the f and g locus allotype markers present on rabbit α-chains revealed that the α-chain is the heavy chain on the Peyer's patch lymphocytes which previously had been shown to be the precursors of IgA-producing plasma cells. In addition, lymphocytes which had been stripped of membrane Ig with pronase and then cultured overnight to allow the sole expression of endogenous membrane Ig were found to have either the µ-chain or the α-chain on their membranes, but not both. These results suggest that most lymphocytes are restricted to the synthesis of one class of heavy chains at a time and that the commitment to synthesizing that particular heavy chain is maintained during the differentiation of lymphocytes into plasma cells. The proportion of lymphocytes with membrane α-chains is higher in the Peyer's patch and appendix, two gut-associated lymphoid tissues (GALT), than in other lymphoid tissues. Since the GALT are enriched sources of precursors for IgA-producing plasma cells compared to nongut-associated tissues, the presence of cells bearing membrane α-chains correlates well with the relative abilities of these tissues to generate IgA plasma cells.
    Type of Medium: Online Resource
    ISSN: 1540-9538 , 0022-1007
    RVK:
    Language: English
    Publisher: Rockefeller University Press
    Publication Date: 1974
    detail.hit.zdb_id: 1477240-1
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  • 4
    Online Resource
    Online Resource
    American Association for the Advancement of Science (AAAS) ; 1971
    In:  Science Vol. 171, No. 3969 ( 1971-01-29), p. 391-394
    In: Science, American Association for the Advancement of Science (AAAS), Vol. 171, No. 3969 ( 1971-01-29), p. 391-394
    Type of Medium: Online Resource
    ISSN: 0036-8075 , 1095-9203
    RVK:
    RVK:
    Language: English
    Publisher: American Association for the Advancement of Science (AAAS)
    Publication Date: 1971
    detail.hit.zdb_id: 128410-1
    detail.hit.zdb_id: 2066996-3
    detail.hit.zdb_id: 2060783-0
    SSG: 11
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  • 5
    Online Resource
    Online Resource
    The American Association of Immunologists ; 1973
    In:  The Journal of Immunology Vol. 111, No. 5 ( 1973-11-01), p. 1595-1597
    In: The Journal of Immunology, The American Association of Immunologists, Vol. 111, No. 5 ( 1973-11-01), p. 1595-1597
    Abstract: In the rabbit, production of b-locus allotypic determinants present on kappa-type light chains can be suppressed by perinatal exposure to anti-b5 antisera (allotype suppression) (1, 2). We have previously demonstrated that b5-bearing lymphocytes are deleted in totally suppressed rabbits with no circulating b5, and that in spontaneous escape from suppression the appearance of b5-bearing lymphocytes immediately precedes the appearance of detectable serum b5 (3). However, during recovery from allotype suppression, the proportion of peripheral blood lymphocytes with membrane b5 approaches normal levels, while circulating b5 levels remain chronically and disproportionately depressed (3). Because there is this unusual discrepancy between the number of b5-bearing lymphocytes and the amount of circulating b5 in rabbits recovering from allotype suppression, and because passive adsorption of circulating b5 molecules to lymphocytes not themselves capable of synthesizing b5 could account for the inappropriately large number of b5-bearing cells, we have investigated these lymphocytes for their ability to synthesize actively their own membrane b5 Ig after enzymatic stripping of the cell surface.
    Type of Medium: Online Resource
    ISSN: 0022-1767 , 1550-6606
    RVK:
    RVK:
    Language: English
    Publisher: The American Association of Immunologists
    Publication Date: 1973
    detail.hit.zdb_id: 1475085-5
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  • 6
    Online Resource
    Online Resource
    Rockefeller University Press ; 1974
    In:  The Journal of Experimental Medicine Vol. 139, No. 3 ( 1974-03-01), p. 581-599
    In: The Journal of Experimental Medicine, Rockefeller University Press, Vol. 139, No. 3 ( 1974-03-01), p. 581-599
    Abstract: Lymphocytes from b5/b9 rabbits were stained in suspension with fluorescent antiallotype antibody reagents to selectively label with fluorescent molecules those cells bearing membrane immunoglobulin (Ig) of the b5 or b9 allotype. After staining, the cells were separated by the fluorescence-activated cell sorter into populations markedly enriched in cells bearing b5 or b9 membrane Ig or totally depleted of cells with detectable membrane Ig. The potential of these separated cells to give rise to Ig-synthesizing plasma cells either in vivo after transfer into irradiated recipients or in vitro during culture in the presence of phytohemagglutinin or pokeweed mitogen was assessed by immunofluorescence. The relative proportion of b5 and b9 cytoplasmic Ig-stained cells (CSC) arising from the separated cells was determined to test directly whether B lymphocytes and their progeny are committed to the synthesis of Ig of one allotype. It was found that b5- and b9-bearing cells gave rise almost exclusively to b5- and b9-producing plasma cells, respectively, in both the in vivo and in vitro assay systems. Most of these CSC were probably not derived from previously existing CSC but arose as the result of the differentiation of lymphocytes with membrane Ig. When cell populations totally depleted of Ig-bearing lymphocytes were cultured, very few CSC were found, indicating that the majority of immediate precursors of CSC have membrane Ig. These results suggest that individual B cell clones are phenotypically restricted to the expression of immunoglobulin genes on one chromosome; the significance of this clonal allelic exclusion is discussed.
    Type of Medium: Online Resource
    ISSN: 1540-9538 , 0022-1007
    RVK:
    Language: English
    Publisher: Rockefeller University Press
    Publication Date: 1974
    detail.hit.zdb_id: 1477240-1
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