THE RETENTION OF <sup>125</sup>I‐IMMUNOGLOBULINS, IgG SUBUNITS AND ANTIGEN‐ANTIBODY COMPLEXES IN RAT FOOTPADS AND DRAINING LYMPH NODES

Herd, Zana L.; Ada, G. L.

doi:10.1038/icb.1969.5

Cited by 28 publications

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“…The IgG molecule is T-shaped with a horizontal length of 14.5 nm (145 X) (FAB2) and a height of 10.7 nm (107 A) (33). The immunoglobulin is probably bound to the cell surface via its Fc piece (12,34). Therefore, in order for cell-surface immunoglobulin molecules not to collide with each other, each one would require a circle of area of diameter slightly greater than the FAB2 distance.…”

Section: Methodsmentioning

confidence: 99%

Antigen-Binding Specificity of Isolated Cell-Surface Immunoglobulin from Thymus Cells Activated to Histocompatibility Antigens

Cone

Sprent

Marchalonis

1972

Proc. Natl. Acad. Sci. U.S.A.

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Lactoperoxidase-catalyzed radioiodination of cell-surface proteins was used in the isolation of cellsurface immunoglobulin from thymus-derived thoracic duct lymphocytes activated to histocompatibility-2 antigens. Immunoglobulin was identified by specific precipitation with antiserum to mouse immunoglobulin prepared in the rabbit and mouse immunoglobulin. Polyacrylamide gel electrophoresis of reduced and alkylated precipitates showed that the immunoglobulin molecules possessed I-type heavy chains and light chains. Cell-surface immunoglobulin isolated from thymus-derived cells activated to histocompatibility-2 antigens possessed binding specificity for the activating antigens.The ability of antisera to immunoglobulin to inhibit the immunocompetence of lymphocytes has been considered to provide evidence that specific interaction of these cells with antigen is mediated by immunoglobulin-like receptors located on the cell surface (1-7). Such studies, however, do not exclude the possibility that receptor sites are not immunoglobulin in nature but are situated sufficiently close to immunoglobulin molecules for antibodies directed against immunoglobulins to block, by steric hindrance, the access of antigen to receptor sites. This objection would be obviated if it were demonstrated that surface immunoglobulin extracted from lymphocytes specifically reactive to a certain antigen could bind specifically to that antigen. Such an approach has become feasible because lactoperoxidase (EC 1.11.1.7)-catalyzed radioiodination (8) of cell surface proteins (9-11) has enabled the isolation and partial characterization of surface immunoglobulin molecules from thymus-influenced (T) lymphocytes (12, 13) and bursa or bone-marrow-derived (B) lymphocytes (11)(12)(13)(14) Sprent and Miller (17). Briefly, 3-month-old (CBA x C57BL)F1 or (CBA x BALB/c)F1 mice were injected intravenously with 2 X 108 CBA thymus cell's within 2 hr after receiving 750 R total body irradiation. 4 Days later the thoracic ducts of the mice were cannulated, and lymph was collected over the first [12][13][14][15][16] (9), except that the total reaction volume was 50 Al. We have previously shown that this technique iodinates cell-surface proteins exclusively but does not affect the viability of the cells (9).After iodination the cells were incubated under short-term cell-culture conditions for 2-6 hr as described (13,18). Under these conditions cell-surface proteins, including immunoglobulin, are released into the cell-culture medium (13, 18). Aliquots were removed at intervals and centrifuged at 40 at 1500 rpm (500 X g) for 10 min, and the supernatants were retained. Radioactive cell-surface immunoglobulin was isolated from the supernatants by specific coprecipitation with antiserum against mouse immunoglobulin prepared in rabbits and purified mouse IgG as carrier (12,13

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Section: Methodsmentioning

confidence: 99%

Antigen-Binding Specificity of Isolated Cell-Surface Immunoglobulin from Thymus Cells Activated to Histocompatibility Antigens

Cone

Sprent

Marchalonis

1972

Proc. Natl. Acad. Sci. U.S.A.

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Transfer of memory cells into antigen‐pretreated hosts. II. Influence of localized antigen on the migration of specific memory B cells

1981

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Following i.v. injection, 2,4,6-trinitrophenol (TNP)-primed memory cells localized in recipient lymph nodes draining a footpad injection of TNP-hemocyanin (TNP-KLH) in greater numbers than in contralateral nodes draining a p-azobenzenearsonate-coupled KLH injection. Such hapten-specific, unilateral memory B cell localization was still observed in immunosuppressed mice when antigen injections were given as long as 4 days prior to the memory cell transfer. The memory cells could be challenged to form plaque-forming cells by footpad injections of TNP-labeled Brucella abortus at 5 days, but not one day, after cell transfer. The present studies further clarify some parameters of this adoptive memory, as a model for the study of persistent local memory. Measures that promoted the unilateral lymph node retention of 125I-labeled antigen also facilitated unilateral accumulation of TNP-specific memory cells. Such measures included pretreatment of the recipients with cyclophosphamide, rather than gamma irradiation, injection of anti-carrier antibody the day before antigen, or use of small doses of preformed immune complexes instead of antigen alone. In general, a high ratio of lymph node-to-spleen and lymph node-to-blood concentration of antigen in recipients appeared crucial for unilateral localization of memory B cells. Splenectomy of recipients prior to cell transfer enhanced the difference in plaque-forming cell responses between draining and contralateral nodes, but decreased their difference when performed 1 day after cell transfer, suggesting that the spleen may have served as a trap for memory cells. I.v. injection of antigen at the time of B cell transfer also interfered with unilateral localization. The results demonstrate that in the presence of persisting depots of antigen within lymph nodes (and absence of significant amounts of antigen elsewhere), memory B cells can be retained locally without activation into antibody-secreting cells. This mechanism may, therefore, by responsible for the phenomenon of local, humoral, immunological memory.

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Isolation of follicular dendritic cells from human tonsils and adenoids. II. Immunocytochemical characterization

et al. 1984

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Follicular dendritic cells (FDC) are specialized cells found only within lymphoid follicles. They bind immune complexes and play a role in the presentation of antigen to follicular B cells and in the generation of B cell memory. In the present report the isolation of FDC from human tonsils and adenoids is described. These isolated cells have an unusual spherical arrangement and enclose lymphocytes within extensions of their membranes. Their ultrastructural features are similar to those observed in situ. The reactivity of isolated FDC with a number of monoclonal antibodies was analyzed by immunofluorescence and by immunostaining (at the electron microscopic level) with colloidal gold. In keeping with the results of previous investigations on tissue sections IgM, IgG and IgA (but not IgD) can be detected on the surface of isolated FDC, as can C3b receptors and the FDC-associated antigen detected by monoclonal antibody R4/23. The immunoglobulins associated with FDC are mostly embedded in an electron-dense material. The majority of the lymphoid cells enclosed within the membrane extensions of FDC are of B cell type. These results suggest that isolated FDC may be suitable for further in vitro investigation of their role in the humoral immune response.

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THE RETENTION OF ¹²⁵I‐IMMUNOGLOBULINS, IgG SUBUNITS AND ANTIGEN‐ANTIBODY COMPLEXES IN RAT FOOTPADS AND DRAINING LYMPH NODES

Cited by 28 publications

References 0 publications

Antigen-Binding Specificity of Isolated Cell-Surface Immunoglobulin from Thymus Cells Activated to Histocompatibility Antigens

Antigen-Binding Specificity of Isolated Cell-Surface Immunoglobulin from Thymus Cells Activated to Histocompatibility Antigens

Transfer of memory cells into antigen‐pretreated hosts. II. Influence of localized antigen on the migration of specific memory B cells

Isolation of follicular dendritic cells from human tonsils and adenoids. II. Immunocytochemical characterization

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THE RETENTION OF 125I‐IMMUNOGLOBULINS, IgG SUBUNITS AND ANTIGEN‐ANTIBODY COMPLEXES IN RAT FOOTPADS AND DRAINING LYMPH NODES

Cited by 28 publications

References 0 publications

Antigen-Binding Specificity of Isolated Cell-Surface Immunoglobulin from Thymus Cells Activated to Histocompatibility Antigens

Antigen-Binding Specificity of Isolated Cell-Surface Immunoglobulin from Thymus Cells Activated to Histocompatibility Antigens

Transfer of memory cells into antigen‐pretreated hosts. II. Influence of localized antigen on the migration of specific memory B cells

Isolation of follicular dendritic cells from human tonsils and adenoids. II. Immunocytochemical characterization

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THE RETENTION OF ¹²⁵I‐IMMUNOGLOBULINS, IgG SUBUNITS AND ANTIGEN‐ANTIBODY COMPLEXES IN RAT FOOTPADS AND DRAINING LYMPH NODES