Quantitative Features of a Sandwich Radioimmunolabeling Technique for Lymphocyte Surface Receptors

Nossal, G. J. V.; Warner, Noel L.; Lewis, Heather; Sprent, J.

doi:10.1084/jem.135.2.405

Cited by 175 publications

(48 citation statements)

References 19 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nevertheless, there is some evidence in the literature that T cells have an Ig receptor (44)(45)(46). Most of these results have been obtained with the use of specific antisera to kappa chains.…”

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

confidence: 95%

Surface Antigens of Immunocompetent Cells

Mond

Takahashi

Thorbecke

1972

The Journal of Experimental Medicine

View full text Add to dashboard Cite

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.

show abstract

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

confidence: 95%

Surface Antigens of Immunocompetent Cells

Mond

Takahashi

Thorbecke

1972

The Journal of Experimental Medicine

View full text Add to dashboard Cite

show abstract

“…Ig-bearing lymphocytes were detected by a sandwich technique using rabbit IgG antimouse Ig followed by Fl-sheep antirabbit Ig essentially as previously described (14) . Labeled cells were counted using a Leitz orthoplan microscope (E .…”

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

View full text Add to dashboard Cite

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.

show abstract

“…The antiserum was prepared in rabbits against an antigen isolated by density gradient sedimentation of the medium from Hodgkin's disease monolayer cultures. In other systems (5)(6)(7)(8), the isotopic antibody method has proven to be a sensitive and quantitative assay of cell surface antigens.…”

Section: Discussionmentioning

confidence: 99%