Antigen Interaction and Heat Inactivation Expose New  Epitopes on Human IgE

Pachlopnik, Jana M.; Stämpfli, Martin R.; Rudolf, Michael P.; Aebischer, Iwan; Kricek, Franz; Miescher, Sylvia; Stadler, Beda M.

doi:10.1159/000024016

Cited by 7 publications

(7 citation statements)

References 16 publications

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“…Thus, detection of antiFcεRI autoAbs that compete with IgE for binding may require lactic acid treatment of the basophils and heat denaturation of sera. Heat denatured IgE is no longer capable of binding to FcεRI and most monoclonal anti-IgE antibodies seem to recognize epitopes that disappear during heat denaturation [50]. For the measurement of serum anti-FcεRI autoAbs that compete with IgE for binding to the receptor it can be useful to denature the serum samples even though it will not fully inactivate complement.…”

Section: Discussionmentioning

confidence: 99%

“…any disturbance of the balance between occupied versus free FcεRI ) contributes to the pathogenesis of autoimmune urticaria is not known. Detection of anti-FcεRI autoAbs in histamine release assays as well as in diagnostic tests based on recombinant FcεRI includes measurement after removal of IgE from the system, either by lactic acid (removal of IgE from cell surface) [56] or by heat denaturation (removal of IgE from sera) [50]. Such assays do not account for the fact that urticaria most often appears on the skin as a local phenomenon and thus a serological profile may not be a true indicator of the in vivo pathology.…”

Section: Discussionmentioning

confidence: 99%

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Natural anti-FcεRIα autoantibodies may interfere with diagnostic tests for autoimmune urticaria

Pachlopnik¹,

Horn²,

Fux³

et al. 2004

Journal of Autoimmunity

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Natural anti-FcεRIα autoantibodies may interfere with diagnostic tests for autoimmune urticaria

Pachlopnik¹,

Horn²,

Fux³

et al. 2004

Journal of Autoimmunity

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“…17 and unpublished observation). However, an anti-C⑀3 mAb (termed 8E7/H8) raised against recombinant C⑀3 was shown to also recognize heat-denatured IgE (41). Thus, it may be speculated that recombinant C⑀3 as well as phage-displayed C⑀3 were not properly folded, thereby adopting a structure similar to heat-denatured IgE.…”

Section: Discussionmentioning

confidence: 99%

Molecular Basis for Nonanaphylactogenicity of a Monoclonal Anti-IgE Antibody

Rudolf

Zuercher

Nechansky

et al. 2000

The Journal of Immunology

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IgE Abs mediate allergic responses by binding to specific high affinity receptors (FcεRI) on mast cells and basophils. Therefore, the IgE/FcεRI interaction is a target for clinical intervention in allergic disease. An anti-IgE mAb, termed BSW17, is nonanaphylactogenic, although recognizing IgE bound to FcεRI, and interferes with binding of IgE to FcεRI. Thus, BSW17 represents a candidate Ab for treatment of IgE-mediated disorders. By panning BSW17 against random peptide libraries displayed on phages, we defined mimotopes that mimic the conformational epitope recognized on human IgE. Two types of mimotopes, one within the Cε3 and one within the Cε4 domain, were identified, indicating that this mAb may recognize either a large conformational epitope or eventually two distinct epitopes on IgE. On the basis of alignments of the two mimotopes with the human IgE sequence, we postulate that binding of BSW17 to the Cε3 region predominantly blocks binding of IgE to FcεRI, leading to neutralization of IgE. Moreover, binding of BSW17 to the Cε4 region may explain how BSW17 recognizes FcεRI-bound IgE, and binding to this region may also interfere with degranulation of IgE sensitized cells (basophils and mast cells). As a practical application of these findings, mimotope peptides coupled to a carrier protein may be used for the development of a peptide-based anti–allergy vaccine by induction of anti-IgE Abs similar to the current approach of using humanized nonanaphylactogenic anti-IgE Abs as a passive vaccine.

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“…Generation and properties of mouse anti–hIgE mAb BSW17 has been described [17]. Anti–hIgE mAbs 4F4 (Cε3 specific) and 11–3 (Cε2 specific) have been described elsewhere [18]. Pc, rabbit anti–mouse IgG1 was obtained from Pharmacia, Uppsala, Sweden, and goat anti–mouse IgG was from BioRad, Hercules, Calif., USA.…”

Section: Methodsmentioning

confidence: 99%

Interaction of Human IgE with Fc Epsilon RI Alpha Exposes Hidden Epitopes on IgE

Nechansky

Ruf

Pursch

et al. 1999

Int Arch Allergy Immunol

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Background: Binding of human IgE via the heavy–chain constant region domain 3 (Cε3) to the α–chain of its high affinity receptor (FcεRIα) is a key event in mediating allergic reactions. We wanted to identify epitopes within Cε3 that are stable to denaturation and to evaluate whether such structures are involved in receptor binding. The existence of stable epitopes would facilitate the generation of compounds that inhibit the IgE–FcεRIα interaction. Methods: Monoclonal anti–human IgE–antibodies against recombinant bacterially synthesized Cε3, which is known to be partly misfolded, were raised in mice. These antibodies were probed for binding to native, immobilized and receptor–bound IgE, respectively, providing tools for the identification of the indicated stable epitopes. Results: Two of the generated antibodies (8E7, 3G9) discriminate between IgE in solution and IgE attached to FcεRIα, pointing towards a steric rearrangement within Cε3 induced upon receptor binding. The described antibodies represent tools for studying the mechanism of the Fcε–FcεRIα interaction and may be of diagnostic value since serum IgE from various human donors was differently recognized by 8E7, which is indicative for naturally occurring IgE molecules with different steric conformation. Conclusion: The presented data support the hypothesis of a conformational change within IgE Cε3 upon receptor binding by showing that monoclonal antibodies raised against recombinant Cε3 differently recognize soluble and receptor–bound IgE. The presence of an IgE portion in sera of human donors that is recognized by 8E7 indicates the existence of IgE molecules in different steric conformations in human blood, which may be related to pathologic parameters.

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Antigen Interaction and Heat Inactivation Expose New Epitopes on Human IgE

Cited by 7 publications

References 16 publications

Natural anti-FcεRIα autoantibodies may interfere with diagnostic tests for autoimmune urticaria

Natural anti-FcεRIα autoantibodies may interfere with diagnostic tests for autoimmune urticaria

Molecular Basis for Nonanaphylactogenicity of a Monoclonal Anti-IgE Antibody

Interaction of Human IgE with Fc Epsilon RI Alpha Exposes Hidden Epitopes on IgE

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