Specific Inactivation of Thymus-derived (T) and Non-thymus-derived (B) Lymphocytes by 125I-labelled Antigen

Basten, Antony; Miller, J. F. A. P.; Warner, Noel L.; Pye, J.

doi:10.1038/newbio231104a0

Cited by 192 publications

(77 citation statements)

References 8 publications

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“…In experiments by Mason and Warner (49) the ability of cells to transfer delayed hypersensitivity in the mouse could be inhibited by previous exposure of the cells to some anti-L chain antisera in vitro in the absence of Cq A similar inhibition was obtained by anti-Ig on the effectiveness of cells to induce graft-vs.-host reactions (50,51). Evidence that immunocompetent thymus cells bind antigen was obtained from "suicide" experiments using highly radioactive protein antigen (13). This phenomenon was inhibited by simultaneous addition of anti-L chain.…”

Section: Complemen Tatio N Of a Nti-o-treated With A N Ti-i G-treatedmentioning

confidence: 66%

Surface Antigens of Immunocompetent Cells

Mond

Takahashi

Thorbecke

1972

The Journal of Experimental Medicine

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The effect of preincubation with anti-θ or anti-mouse immunoglobulin (Ig) and complement (C') on immune responsiveness of spleen cells from BALB/c mice immunized with sheep erythrocytes (SE) was investigated. Both treatments greatly depressed the remaining ability to produce a secondary response to SE in vitro. Normal BALB/c spleen cells were far less effective in reconstituting the responses of such depleted cell populations than were much smaller numbers of untreated immune spleen cells. Thymus-derived cell (T cell) memory appeared early after immunization and showed specificity for the immunizing antigens. Recombination of anti-Ig-treated with anti-θ-treated immune spleen cells resulted in virtually complete reconstitution of responsiveness. The presence of immunological memory in T cells and the nature of their surface receptors are discussed.

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Section: Complemen Tatio N Of a Nti-o-treated With A N Ti-i G-treatedmentioning

confidence: 66%

Surface Antigens of Immunocompetent Cells

Mond

Takahashi

Thorbecke

1972

The Journal of Experimental Medicine

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“…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).…”

Section: Introductionmentioning

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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“…Antigen-induced suicide of T and B cells was carried out by a modification of the method of Basten et al . (18) . For T cells, 10 jig " 6 I-labeled F'YG (sp act 150 pCi/,Ug) was added to 5 x 10 8 anti-B serum-treated spleen cells from mice primed to FyG and HRBC for 30 min at 4°C .…”

mentioning

confidence: 99%

“…Excess radioactivity was removed by centrifugation through two discontinuous FCS gradients. B-cell suicide was carried out by incubating anti-B serum-treated spleen cells primed to FyG and HRBC with 'a 'Llabeled F-yG entirely in the cold as previously described (18) . For inhibition studies, purified T cells or anti-B serum-treated spleen (B) cells were incubated with anti-H-2k Fab (1 mg/108 cells) for 3 h at 4°C.…”

mentioning

confidence: 99%

Relationship between Fc receptors, antigen-binding sites on T and B cells, and H-2 complex-associated determinants.

Basten¹,

Miller²,

Abraham³

1975

The Journal of Experimental Medicine

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The relationship between H-2 complex-associated determinants, Fc receptors, and specific antigen-recognition sites on T and B cells was examined by binding and functional assays. The Fc receptor was detected by radiolabeled immune complexes or aggregated human IgG. Both these reagents selectively bound to B cells, not to T cells. When spleen cells, from mice primed to several antigens, were exposed to highly substituted radioactive aggregates, their capacity to transfer both a direct and indirect plaque-forming cell response to these antigens was abrogated. Addition of B cells, but not of T cells, restored responsiveness. Complexed Ig binding to Fc receptors was prevented by pretreatment of mixed lymphoid cell populations with antisera directed against membrane components on the same cell (e.g., H-2) and on other cells (e.g., theta). The lack of specificity of inhibition was thought to be due to the formation on cell surfaces of antigen-antibody complexes which would then attach to the Fc receptor during the incubation precedure. Specific blockade of the Fc receptor during the incubation procedure. Specific blockade of the Fc receptor however occurred when B cells were pretreated with the Fab fragments of anti-H-2 antibody. This was demonstrated autoradiographically and by inhibition of aggregate-induced suicide. The blocking activity of ante-H-2 Fab was removed by absorption with spleen cells from thymectomized irradiated mice but not with thymus cells of appropriate specificity. This suggested that the antibodies involved had specificity for determinants on the B-cell membrane distinct from those coded by the K or D end of the H-2 complex, and either absent from, or poorly represented on, thymus cells. Specific antigen-induced suicide of B cells was achieved simply by incubating the cells with radioactive antigen in the cold. T-cell suicide on the other hand required that the 125I-labeled antigen be presented to the T cells at 37 degrees-C on the surface of spleen cells from antigen-primed mice. Pretreatment of T cells with the Fab fragment of anti-H-2 antibody protected them from the suicide effect. By contrast no such protection of B cells could be achieved by this procedure. In other words H-2 (? Ir)-associated determinants may not only be in close proximity to the antigen-binding site on T cells but, in addition, may be involved in the effective operation of the receptor.

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Specific Inactivation of Thymus-derived (T) and Non-thymus-derived (B) Lymphocytes by 125I-labelled Antigen

Cited by 192 publications

References 8 publications

Surface Antigens of Immunocompetent Cells

Surface Antigens of Immunocompetent Cells

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

Relationship between Fc receptors, antigen-binding sites on T and B cells, and H-2 complex-associated determinants.

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