Characterization of the target cell receptor for IgE—I. Solubilization of IgE-receptor complexes from rat mast cells and rat basophilic leukemia cells

Conrad, Daniel H.; Berczi, István; Froese, Arnold

doi:10.1016/0019-2791(76)90343-8

Cited by 56 publications

(42 citation statements)

References 21 publications

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“…The reaginic antibodies were identified as belonging exclusively to the IgE class of immunoglobulins on the basis of the follow ing criteria: (i) PCA reactions were elicited 48 h after the passive sensitization of rats with the reaginic sera; (ii) these PCA reac tions were abolished by heating the reaginic sera at a temperature of 56 °C for a period of 4 h prior to use for passive sensitization of rats, and (iii) the PCA activity of the rat reaginic sera was removed by coprecipita tion (which was kindly performed by Dr. D. H. Conrad of this Department) of the IgE immunoglobulins with a rabbit antiserum specific for a rat myeloma IgE [3], followed by the addition of a goat antiserum to rabbit gammaglobulins [5].…”

Section: Resultsmentioning

confidence: 99%

Modulation of the IgE Antibody Response in Rats to Kentucky Blue Grass Pollen Allergens

Ekramoddoullah¹,

Kisil²,

Sehon³

1977

Int Arch Allergy Immunol

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The low molecular weight dialyzable fraction (D) prepared from the aqueous extract of Kentucky Blue Grass pollen was shown to suppress the formation of IgE antibodies in rats immunized with the nondialyzable fraction (R). In an attempt to establish the nature of the constituents responsible for this suppression, D was fractionated by gel filtration through Sephadex G-25. The first fraction eluted (D_I) elicited skin reactions in rats sensitized with a rat reaginic serum to R and also gave a precipitate with a rabbit antiserum to R. A later fraction (D_III) was devoid of these two properties. To investigate the effects of these fractions on the antibody response, rats received either D_I or D_III, administered in saline, prior to their immunization with R in presence of aluminum hydroxide. Pretreatment with D_I resulted in a reduction of IgE antibody levels as compared with the IgE antibody response in control animals which had received pretreatment only with saline; however, pretreatment with D_I did not affect the anti-R-hemagglutinating titers. In contrast, pretreatment with D_III enhanced both the IgE and the hemagglutinating antibodies. Hence, it is concluded that the composite fraction D contains one group of constituents capable of suppressing and another of enhancing the IgE antibody response.

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Section: Resultsmentioning

confidence: 99%

Modulation of the IgE Antibody Response in Rats to Kentucky Blue Grass Pollen Allergens

Ekramoddoullah¹,

Kisil²,

Sehon³

1977

Int Arch Allergy Immunol

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“…It is a tetramere of one IgEbinding a chain (2,3), one fi chain (4,5), and two identical disulfide-linked y chains (6,7). Complementary DNA clones for the rat a (8,9), , (10), and y ( 11) and for the human a (9, 12) have been isolated.…”

Section: Introductionmentioning

confidence: 99%

Early expression of high-affinity receptor for immunoglobulin E (Fc epsilon RI) during differentiation of mouse mast cells and human basophils.

Thompson

Metcalfe²,

Kinét³

1990

J. Clin. Invest.

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“…Elmhurst, Ill.) in 0.2 M borate-buffered saline, pH 8.0, as described previously (2,6). When not used immediately, preparations were stored at -90°.…”

Section: Methodsmentioning

confidence: 99%

“…That the receptor can be iodinated (2), that radioactive amino acids and sugars can be incorporated into it (3), that it is sensitive to proteases (2,3) and moderate changes in temperature and pH (4)-all suggest it is a glycoprotein. Its activity is unchanged by exposure to a variety of lipases (G. Rossi, unpublished observations).…”

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confidence: 99%

Molecular weight and valence of the cell-surface receptor for immunoglobulin E.

Newman¹,

Rossi²,

Metzger³

1977

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

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The molecular weight of the active solubilized cell-surface receptor for immunoglobulin E (IgE) was measured in nonionic detergent. The diffusion coefficient was estimated by gel filtration, the partial specific volume was estimated from the differential sedimentation in sucrose gradients prepared from H20 and D20, and the sedimentation constant was estimated from the same centrifugation experiments. receptors were assessed using the (NH4)2SO4 assay described previously (6). Gel Filtration. Gel exclusion chromatography was performed on 92 X 1.4 cm columns of Sepharose 6B (Pharmacia, Piscataway, N.J.) equilibrated at 40 with borate buffer containing 0.5% or 1% Nonidet P-40 and 0.05% bovine serum albumin'. Column fractions of 1% bed volume were collected and flow rates were -<1 bed volume/5 hr. The exclusion volume (V0) was marked with '311-labeled tobacco mosaic virus (a gift from T. Triche, National Cancer Institute) after we observed that dextran blue appeared to interact with the solubilized receptor. A standard included volume (VG) was marked with the IgG and the RF for the unknowns and standards was calculated from their elution volumes (Ve) using the formula RF = (Ve -VO)/(VG -VO) [ 1]The RF of each standard was plotted versus 1/D20o, and the line fit by least squares was used to convert the RF values of the unknowns to their D2o0, values (15).Sucrose Density Gradient Centrifugation. Centrifugation experiments were performed with an L265B-G ultracentrifuge using an SW 40 rotor (Beckman Instrument Co., Palo Alto, Calif.). Pairs of 14.2 ml, 2% to 8% (wt/vol) linear sucrose gradients in borate buffer containing 1% Nonidet P-40 and 0.05% bovine serum albumin were prepared by pumping through mixing chambers into nitrocellulose tubes. After storage for -2 hr at 40, Al of sample in a solvent which was identical to that of the gradient except for the sucrose was layered on the gradient and the tubes were centrifuged at 40,000 rpm for 14-20 hr at 4°. Fractions of approximately 250,u (7 drops) were collected automatically. There was less than 5% variation in sample size between the second and last fraction collected and refractive index measurements on unspun gradients showed excellent linearity. For the IgE, IgE-receptor complex, and marker proteins the radioactivity of the samples could be counted directly. For the free receptor 75-140,ul portions of the fractions were diluted at least 2:1, incubated with labeled IgE, and assayed with the (NH4)2SO4 technique. The densities of the starting solvents were measured directly in a 10 or 25 ml pycnometer and the refractive index for each was determined

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Characterization of the target cell receptor for IgE—I. Solubilization of IgE-receptor complexes from rat mast cells and rat basophilic leukemia cells

Cited by 56 publications

References 21 publications

Modulation of the IgE Antibody Response in Rats to Kentucky Blue Grass Pollen Allergens

Modulation of the IgE Antibody Response in Rats to Kentucky Blue Grass Pollen Allergens

Early expression of high-affinity receptor for immunoglobulin E (Fc epsilon RI) during differentiation of mouse mast cells and human basophils.

Molecular weight and valence of the cell-surface receptor for immunoglobulin E.

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