Effect of heating at 56°C on mouse IgE antibodies

Prouvost‐Danon, Annie; Binaghi, R; Abadie, A

doi:10.1016/0019-2791(77)90284-1

Cited by 34 publications

(21 citation statements)

References 14 publications

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“…Sera were two fold serially diluted with saline and injected in a volume of 10,u] into ddY mouse ear. When assaying IgG1 anti body titer, sample sera were heated at 56°C for 2 hours before dilution (19,25). For detecting IgG1 antibody, mice were challenged by intravenous injection of 0.25 ml of 0.5% Evans blue saline solution containing 0.25 mg of antigen 1.5 hours after the sensitization.…”

Section: Serum Antibody Titermentioning

confidence: 99%

Active Cutaneous Anaphylaxis (ACA) in the Mouse Ear

Inagaki

Miura

Nagai

et al. 1992

Japanese Journal of Pharmacology

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ABSTRACT-Active cutaneous anaphylaxis (ACA) was studied in the ear of female BALB/c mice. Mice were immunized with ovalbumin in the presence of aluminium hydroxide gel or complete Freund's adjuvant (CFA). Two weeks after the immunization, ACA was elicited in the mouse ear by injecting 10,ul of antigen solution intradermally into the ear lobe. ACA was assessed by the amount of extravasated dye, which was given intravenously just after the antigen injection. Antiallergic drugs (tra nilast, ketotifen and azelastine), antihistamines (chlorpheniramine, diphenhydramine and mequitazine), fl-stimulants (isoproterenol and salbutamol), theophylline and glucocorticoids (hydrocortisone, predni solone and dexamethasone) inhibited the reaction significantly. These drugs inhibited both ACA in mice immunized with alum-precipitated antigen and ACA in mice injected with CFA-emulsified antigen similarly. ACA in the mouse ear might be a useful tool for studying drugs for allergy.Keywords: Active cutaneous anaphylaxis, Ear (mouse), Antiallergic drug, IgE antibody, IgG1 antibody Passive cutaneous anaphylaxis (PCA) (1, 2) is a use ful experimental model for studying drugs for allergy (3 6). The rat has been the most useful species for this purpose and the reaction is assessed by measuring the amount of extravasated dye (7,8). On the other hand, the mouse is the most established experimental animal and there are many strains of mice with unique im munological characteristics (9-12).Furthermore, Feinberg (13) reported the suitability of mouse ear for induction of PCA, and Prouvost-Danon and Binaghi (14) demonstrated that mouse ear had higher reactivity for PCA than mouse dorsal skin. According to these re ports, we established a method for PCA in the mouse ear by assessing the amount of extravasated dye (15) and characterized it pharmacologically and immunologi cally (16-23). However, although passive sensitization provides a simple model that is suitable for analyzing the mechanisms involved in both allergic reactions and drug action, active sensitization is also important for studying allergic diseases, because allergic diseases are caused by active sensitization. In the present study, therefore, we applied the technique of mouse ear PCA to active cutaneous anaphylaxis (ACA) elicited in female BALB/c mice, and examined the effect of drugs which are used for allergic diseases on the ACA. MATERIALS AND METHODS MiceFemale BALB/c mice were used for ACA and male ddY mice were used for titration of serum antibodies. Both mice were purchased from Japan SLC, Inc. (Hamamatsu, Japan). Antigen and adjuvantsOvalbumin (OA, 5 times crystalized) was purchased from Seikagaku Kogyo Co., Ltd. (Tokyo, Japan). As adjuvants, aluminium hydroxide gel (alum) and com plete Freund's adjuvant (CFA) were used. Alum was prepared according to the method of Levine and Vaz (24), and CFA was obtained from Nacalai Tesque, Inc. (Kyoto, Japan). ACATen-weeks-old female BALB/c mice were immunized with OA in the presence of an adjuvant. To induce IgE antibody mainly, 1,ug...

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Section: Serum Antibody Titermentioning

confidence: 99%

Active Cutaneous Anaphylaxis (ACA) in the Mouse Ear

Inagaki

Miura

Nagai

et al. 1992

Japanese Journal of Pharmacology

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“…The incubation of IgE at 56°C for a few hours significantly reduces anaphylactic activity, triggered by the binding of Fc of IgE to the Fcε receptor, but does not have a notable effect on the antigen-binding activity in the Fab region (Prouvost-Danon et al 1977;Binaghi and Demeulemester 1983;Ishizaka et al 1986). Under the same experimental conditions, anaphylactic IgG (IgG2a) did not lose its activity (Bloch et al 1968), indicating the difference in heat resistance between the Fc domains of IgE and IgG.…”

Section: Heat-induced Denaturation Of Iggmentioning

confidence: 99%

Heat denaturation of the antibody, a multi-domain protein

2017

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The antibody is one of the most well-studied multi-domain proteins because of its abundance and physiological importance. In this article, we describe the effect of the complex, multi-domain structure of the antibody on its denaturation by heat. Natural antibodies are composed of 6 to 70 immunoglobulin fold domains, and are irreversibly denatured at high temperatures. Although the separated single immunoglobulin fold domain can be refolded after heat denaturation, denaturation of pairs of such domains is irreversible. Each antibody subclass exhibits a distinct heat tolerance, and IgE is especially known to be heat-labile. IgE starts unfolding at a lower temperature compared to other antibodies, because of the low stability of its CH3 domain. Each immunoglobulin domain starts unfolding at different temperatures. For instance, the CH3 domain of IgG unfolds at a higher temperature than its CH2 domain. Thus, the antibody has a mixture of folded and unfolded structures at a certain temperature. Co-existence of these folded and unfolded domains in a single polypeptide chain may increase the tendency to aggregate which causes the inactivation of the antibody.

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“…It has been shown that mouse IgE binds with high affinity to rat mast cell Fce receptors (25,26). Rat mast cells incubated with mouse IgE under the conditions described above later bound an average of 24 If one assumes the relative yield of unoccupied receptors to be 100%, the receptors that had been occupied by mouse IgE were isolated with a relative efficiency of 68%.…”

Section: Selective Phosphorylation Of Ige Receptor a Componentmentioning

confidence: 99%

Selective phosphorylation of the IgE receptor in antigen-stimulated rat mast cells.

Hempstead¹,

Parker²,

Kulczycki³

1983

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

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Purified rat serosal mast cells were sensitized with mouse immunoglobulin E (IgE) anti-2,4-dinitrophenyl antibody, partially depleted of phosphate, labeled with [32P]orthophosphate, and stimulated with dinitrophenylated bovine serum albumin or control protein. After 15-120 seconds at 37C, the cells were extracted with nonionic detergent. IgE receptors were purified by repetitive affinity chromatography and were analyzed by NaDodSO4/polyacrylamide gel electrophoresis and radioautography. Antigenic stimulation of intact rat mast cells produced a rapid and marked increase in the phosphorylation of the surface-exposed a component of the IgE receptor. However, phosphorylation of the 33,000 Mr (3 component of the IgE receptor was not altered significantly by antigen stimulation. This suggests that the selective increase in phosphorylation of the IgE receptor a component may be part of the physiologic mediator secretion process triggered by antigen.Mast cells and basophils have cell-surface receptors that bind IgE with high affinity. Each IgE receptor is composed of a surface-exposed glycoprotein (1, 2), referred to as the a component in recent terminology (3), and a nonglycosylated protein (4, 5) termed the ,B component. Tryptic peptide mapping studies indicate that each component is a distinct polypeptide (6). The a component of rat mast cells, Mr 45,000 (7), is somewhat smaller than the a component of rat leukemic basophils analyzed in the same NaDodSO4/polyacrylamide gel system (6), whereas P components of both cell types are indistinguishable in size (Mr 33,000) (6, 7).The apposition of adjacent receptors on rat mast cells is the necessary and sufficient signal (8) for cell activation leading to the noncytotoxic release and generation of a variety of cellular mediators. Mast cell activation has been associated with changes in diacylglycerol levels (9), phospholipid methylation (10), cyclic AMP levels (11), influx of calcium (10), and protein phosphorylation (12). We have demonstrated that the IgE receptor a component of rat mast cells is a phosphoprotein and that its phosphorylation is significantly increased by a nonimmunologic stimulus of histamine release (7). In this study, we demonstrate that an immunologic stimulus with antigen produces a rapid and selective increase in phosphorylation of the a component of the rat mast cell IgE receptor. Thus, it seems possible that increased phosphorylation of the receptor a component is part of the antigen-stimulated process leading to mediator secretion. MATERIALS AND METHODSAntigen and IgE Antibody. The monoclonal 2,4-dinitrophenyl (DNP)-specific murine IgE antibody H1 DNP-E-26 (13) was isolated from ascites from the hybridoma made available by Fu-Tong Liu and David Katz. Protein soluble in 30% saturated ammonium sulfate was precipitated by 60% saturated ammonium sulfate (13) and was further purified by gel filtration on Sepharose CL-6B. Based on polyacrylamide gel electrophoresis in NaDodSO4, the IgE was >95% pure. DNP-treated bovine serum albumin (DNP-albumin...

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Effect of heating at 56°C on mouse IgE antibodies

Cited by 34 publications

References 14 publications

Active Cutaneous Anaphylaxis (ACA) in the Mouse Ear

Active Cutaneous Anaphylaxis (ACA) in the Mouse Ear

Heat denaturation of the antibody, a multi-domain protein

Selective phosphorylation of the IgE receptor in antigen-stimulated rat mast cells.

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