Evidence for independent genetic regulation of the expression of different antibody classes in anti‐sheep red blood cell responses

Séman, Michel; Chevalier, Florence; Stanislawski, M.

doi:10.1002/eji.1830080409

Cited by 22 publications

(8 citation statements)

References 27 publications

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“…After washing with 10 mM Tris-HCl/1.5 mM EDTA, pH 7.4/30% glycerol buffer (10 ml) and with the same buffer but containing 0.1 M NaCl (10 ml) and finally 5 mM pH 7.4 sodium phosphate buffer (10 ml) the receptor was eluted in 0. 8 Buttin et al. (6).…”

Section: Methodsmentioning

confidence: 99%

Monoclonal antibodies to rabbit progesterone receptor: crossreaction with other mammalian progesterone receptors.

Logeat¹,

Hai²,

Fournier³

et al. 1983

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

133

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A mouse was immunized with purified rabbit uterine cytosolic progesterone receptor (specific activity: 3 nmol of steroid bound per mg of protein). After fusion of its spleen cells with Sp2-OAg myeloma cells, supernatants of 11 hybrid cultures were found to react in both an immunoenzymatic test and a double-immunoprecipitation test with the progesterone receptor. Clones were obtained from the five hybrid cells that gave the strongest response in both tests. Antibodies from cell culture supernatants and ascitic fluids were characterized. Three are of the IgGl and two of the IgG2a isotype. Their apparent affinity for the progesterone receptor was measured by immunoprecipitation in physiological salt conditions. The equilibrium dissociation constants were between 0.1 and 4 nM. All five monoclonal antibodies crossreacted with the rabbit nuclear receptor, the human cytosolic receptor, and other mammalian (rat, guinea pig) but not avian (chicken) cytosolic progesterone receptors. There was no interaction with the glucocorticoid receptor and corticosteroid binding globulin.The study of steroid hormone receptors has been hampered for many years by difficulties in the purification of these proteins and by the impossibility of using immunological tools for their detection and quantification. Initial progress in this field has been made for the estrogen receptor (1, 2). In the case of the progesterone receptor, preparation of polyclonal antibodies against mammalian receptors (3) and recently against avian receptors (4) has been reported. However for such antigens, which occur in low concentrations and are difficult to purify, the possibility remains of misinterpretations caused by the presence of antibodies directed against proteins contaminating the receptor preparation.To solve this problem we have undertaken the preparation of monoclonal antibodies against the rabbit progesterone receptor. MATERIALS AND METHODSPurification of the Rabbit Uterine Progesterone Receptor. Receptor was purified as described (3). However, to concentrate the receptor and increase the purity a final purification step was added. Receptor eluted from the hydroxyapatite column with 0.2 M sodium phosphate, pH 7.4/30% (vol/vol) glycerol buffer was diluted 1:3.2 in 1 mM sodium phosphate, pH 7.4/30% glycerol buffer. It was applied to a small (0.7-ml) calf thymus DNA-cellulose column. After washing with 10 mM Tris-HCl/1.5 mM EDTA, pH 7.4/30% glycerol buffer (10 ml) and with the same buffer but containing 0.1 M NaCl (10 ml) and finally 5 mM pH 7.4 sodium phosphate buffer (10 ml) the receptor was eluted in 0.8 ml of 5 mM.sodium phosphate/0.5 M NaCl buffer, pH 8.3. The specific activity of the receptor preparation that was used for immunization was 3 nmol of steroid bound per mg of protein. Receptor concentration was 750 pmol/ 0.8 ml.Immunization. A 3-month-old BALB/c mouse received subcutaneous injections of 375 pmol of receptor. The receptor solution was concentrated 2-fold by lyophilization and emulsified with an equal volume (0.2 ml) of complet...

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Section: Methodsmentioning

confidence: 99%

Monoclonal antibodies to rabbit progesterone receptor: crossreaction with other mammalian progesterone receptors.

Logeat¹,

Hai²,

Fournier³

et al. 1983

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

133

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“…Previous results with C3H/SW and CWB/13 H-2 b congenic strains (Seman et al 1978) suggested that low IgG2a response is not linked to this H-2 haplotype. We have further investigated this question by analyzing the segregations of 11-2 haplotypes and IgG2a responses in F 2 and F z x BALB/c animals.…”

Section: Absence Of Linkage With H-2 Haplotypesmentioning

confidence: 94%

“…Several arguments led us to conclude that there is an independent genetic control over the expression of the three major IgG isotypes in the response to SRBC. Previous experiments with C3H/Sw and CWB/13 congenic fines have also suggested that genes involved in the regulation of the expression of different IgG subclasses might not be linked to the 1-1-2 locus or with CH allotypes (Seman et al 1978).…”

Section: Introductionmentioning

confidence: 95%

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Genetic control of igG2a production in the response to sheep erythrocytes

Séman¹,

Zilberfarb²,

Stanislawski

et al. 1978

Immunogenetics

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A low IgG2a response in B10 mice during the primary response to sheep red blood cells (SRBC) is described. Analysis of the response in B10 × BALB/c hybrid progenies and in congenic strains indicates that this low response is a dominant phenotype placed under the control of a single Mendelian gene or a group of closely linked genes. This gene(s) is neither linked to CH allotypes orH2 haplotypes, nor is it sex-linked. It can be considered as an isotype- and antigenspecific regulatory gene of the immune response.

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“…The functional specificity of these reagents has been verified in splenic plaque assays [18] and by staining of mouse myeloma cells [19]. The final characteristics of the antisera are given in Table 1.…”

Section: Other Antiseramentioning

confidence: 99%

Antibody response to embryonal carcinoma cells in syngeneic mice

et al. 1979

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The immunoglobulin (Ig) classes and subclasses of the specific antibodies contained in antisera raised in male 129/Sv mice against cells of a syngeneic clonal line of embryonal carcinoma (F9) have been determined. Cytotoxic activity was found associated almost exclusively with anti-F9 IgM (micro kappa) antibodies. A large part of anti-F9 activity was found associated with IgG1 (gamma 1 kappa and presumably also gamma 1 lambda) antibodies, and was detectable only by direct immunofluorescence. Traces of specific IgG2a and IgG2b antibodies were also found. No IgG3 and IgA antibodies reacting with embryonal carcinoma cells were detectable under these conditions. The serum of F9 tumor-bearing animals had a very similar Ig composition. Furthermore, IgM and IgG1, but not IgG2 antibodies, were detected at the surface of in vivo growing F9 tumor cells.

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Evidence for independent genetic regulation of the expression of different antibody classes in anti‐sheep red blood cell responses

Cited by 22 publications

References 27 publications

Monoclonal antibodies to rabbit progesterone receptor: crossreaction with other mammalian progesterone receptors.

Monoclonal antibodies to rabbit progesterone receptor: crossreaction with other mammalian progesterone receptors.

Genetic control of igG2a production in the response to sheep erythrocytes

Antibody response to embryonal carcinoma cells in syngeneic mice

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