The relationship between surface immunoglobulin isotype and immune function of murine B lymphocytes II. Surface immunoglobulin isotopes on unprimed B cells in the spleen.

Zan‐Bar, Israel; Vitetta, Ellen S.; Strober, Samuel

doi:10.1084/jem.145.5.1206

Cited by 45 publications

(24 citation statements)

References 28 publications

(14 reference statements)

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“…The experimental protocol used to prepare the anti-mause S serum. The procedure was adapted from that described by Abney et al (1976) and Zan-Bar et al (1977a). gg VITETTA & UHR consistent with the results of negative selection in so far as y'* cells generate IgG respcmses, whereas (i cells are predominantly precursors of IgM secretion.…”

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IgD and B Cell Differentiation

Vitetta

Uhr

1977

Immunological Reviews

115

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SDS-PAGE= scxlium dodecyl sulfate polyacrylamide gel electrophoresis FACS = fluorescent activated cell sorter PFC = plaque-forming cell FCS = fetal calf serum BSA = bovine serum albumin C = complement LPS = lipopolysaccharide TD = thymus-dependent TI = thymus-independent ' SRBC = sheep red blood cells ' CR = complement receptorThe discovery of murine IgDThe early biochemical studies of cell surface immunoglobulin (Ig) on murine lymphocytes revealed that IgM in its monomeric form was the major cell surface Ig (Vitetta et al. 1971, Marchalonis et al. 1972). Thus, after enzymatic radioiodination of intact splenocytes, cell lysis, precipitation with anti-^, reduction of the dissolved precipitate, and electrophoresis on sodium dodecyl suifate (SDS-PAGE), peaks with the mobility of fi and L chains were obtained. If the supernatant was precipitated on the following day with anti-Ig, no further immunoglobulin was demonstrated by SDS-PAGE examination of this second precipitation. In later experiments, anti-Ig precipitates were electrophoresed on acrylamide gels of increasing acrylamide concentration in an effort to detect subpopulations of /a chains containing different amounts of carbohydrate. When higher concentrations of gels were used, it appeared that the H chain peak obtained after precipitation with anti-Ig could be separated into two peaks, one of which had a faster electrophoretic mobility and appeared more heterogeneous than the

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“…Two sorting experiments were carried out for each immunoglobulin Isotype. In each experiment, a group of mice received a given dose of bright or dull cells (Zan-Bar et al 1977a…”

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confidence: 99%

IgD and B Cell Differentiation

Vitetta

Uhr

1977

Immunological Reviews

115

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“…An alternative explanation is that maintenance of IgD expression may rescue the cells undergoing programmed cell death. However, the role of IgM and IgD in tolerance induction has remained a matter of controversy [12][13][14][15][16][17]. Mice deficient for IgD show a reduced B cell compartment, with 30-50% fewer B cells in the spleen and lymph nodes, indicating that IgD may be important in homeostasis of B cells [18].…”

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The absence of immunoglobulin D B cell receptor-mediated signals promotes the production of autoantibodies and exacerbates glomerulonephritis in murine lupus

Guo

Tian

Guo

et al. 2011

Clinical and Experimental Immunology

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“…jj+b+ cells appear to be the major precursors of the primary IgM antibody response to thymus-dependent (TD) antigens (8,9). The roles played by each of the two isotypes on such cells are not known.…”

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Blocking of primary in vitro antibody responses to thymus-independent and thymus-dependent antigens with antiserum specific for IgM or IgD.

Cambier

Ligler

Uhr

et al. 1978

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

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The effect of anti-M and anti-6 on the primary in vitro IgM response of murine splenocytes to thymus-dependent (trinitrophenylated erythrocytes) and thymus-independent (trinitrophenylated brucella) forms of trinitrophenyl was studied. The results indicate that either anti-M or anti-S can block the response of adult splenocytes to the thymus-dependent antigen. The thymus-dependent responses of neonatal splenocytes that bear a low concentration of IgD were also abrogated by treatment with anti-S. In contrast, anti-,u, but not anti-5, blocked the response of adult splenocytes to the thymus-independent antigen used. These results indicate that both IgM and IgD are receptors required for triggering cells by a thymus-dependent antigen but that only IgM receptors are required for triggering by the thymus-independent antigen used. MATERIALS AND METHODSAntisera. The preparation and specificity of rabbit antimouse Ig (anti-Ig), rabbit anti-,u, and goat anti-rabbit IgThe costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisemnent" in accordance with 18 U. S. C. §1734 solely to indicate this fact.(GARIg) have been described (17). Rabbit anti-6 was judged to be monospecific by published criteria (18), including: (i) a single peak on sodium dodecyl sulfate/polyacrylamide gel electrophoresis after reaction with a lysate of iodinated splenocytes; (ii) immunofluorescent staining of the predicted numbers of cells from various lymphoid tissues; (iii) inability to stain splenocytes after their treatment with either anti-Ig or allotypic (19) anti-6; and (iv) independent capping of surface molecules on splenocytes with anti-6 and anti-A. Chromatographically purified fractions of normal rabbit serum (NRS), anti-,q, anti-Ig, or a 30% (NH4)2SO4 saturation precipitate of anti-6 was used for in vitro cell culture. The IgG fraction from GARIg was conjugated with fluorescein isothiocyanate (Sigma) (FITC-GARIg) and chromatographed on DEAE-cellulose (Whatman DE52) to give a fraction with molar fluorescein/ protein ratio of 2.6.Capping of Cell Surface Ig. Splenocytes were incubated for 30 min at 4°with NRS or the rabbit antisera described above. Cells were washed and exposed to GARIg (20) for 90 min at 370 in complete medium to achieve capping.Immunofluorescent Staining. Viable cells were prepared and treated with NRS or antisera specific for Ig, ,u, or 6 as described (20 Preliminary experiments had indicated that capping alone with any of the anti-Igs used was ineffective in blocking subsequent immune responses, incubation with anti-Igs only (without prior capping) was marginally effective. Therefore, in the experiments described, control cells were capped with

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The relationship between surface immunoglobulin isotype and immune function of murine B lymphocytes II. Surface immunoglobulin isotopes on unprimed B cells in the spleen.

Cited by 45 publications

References 28 publications

IgD and B Cell Differentiation

IgD and B Cell Differentiation

The absence of immunoglobulin D B cell receptor-mediated signals promotes the production of autoantibodies and exacerbates glomerulonephritis in murine lupus

Blocking of primary in vitro antibody responses to thymus-independent and thymus-dependent antigens with antiserum specific for IgM or IgD.

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