Determination of Allergen Specificity by Heavy Chains in Grass Pollen Allergen-specific IgE Antibodies

Overview

Journal J Allergy Clin Immunol

Specialty Allergy & Immunology

Date 2012 Dec 5

PMID 23206656

Citations 5

Authors

Elisabeth Gadermaier

Sabine Flicker

Christian Lupinek

Peter Steinberger

Rudolf Valenta

Affiliations

Soon will be listed here.

Abstract

Background: Affinity and clonality of allergen-specific IgE antibodies are important determinants for the magnitude of IgE-mediated allergic inflammation.

Objective: We sought to analyze the contribution of heavy and light chains of human allergen-specific IgE antibodies for allergen specificity and to test whether promiscuous pairing of heavy and light chains with different allergen specificity allows binding and might affect affinity.

Methods: Ten IgE Fabs specific for 3 non-cross-reactive major timothy grass pollen allergens (Phl p 1, Phl p 2, and Phl p 5) obtained by means of combinatorial cloning from patients with grass pollen allergy were used to construct stable recombinant single chain variable fragments (ScFvs) representing the original Fabs and shuffled ScFvs in which heavy chains were recombined with light chains from IgE Fabs with specificity for other allergens by using the pCANTAB 5 E expression system. Possible ancestor genes for the heavy chain and light chain variable region-encoding genes were determined by using sequence comparison with the ImMunoGeneTics database, and their chromosomal locations were determined. Recombinant ScFvs were tested for allergen specificity and epitope recognition by means of direct and sandwich ELISA, and affinity by using surface plasmon resonance experiments.

Results: The shuffling experiments demonstrate that promiscuous pairing of heavy and light chains is possible and maintains allergen specificity, which is mainly determined by the heavy chains. ScFvs consisting of different heavy and light chains exhibited different affinities and even epitope specificity for the corresponding allergen.

Conclusion: Our results indicate that allergen specificity of allergen-specific IgE is mainly determined by the heavy chains. Different heavy and light chain pairings in allergen-specific IgE antibodies affect affinity and epitope specificity and thus might influence clinical reactivity to allergens.

Citing Articles

Rhubarb-Evoke Mucus Secretion through Aggregation and Degranulation of Mast Cell in the Colon of Rat: In vivo and ex vivo studies.

Wu D, Xue X, Gao C, Liu Y, Wang T, Li L Sci Rep. 2019; 9(1):19375.

PMID: 31852975 PMC: 6920142. DOI: 10.1038/s41598-019-55937-7.

On the Lipophilic Nature of Autoreactive IgE in Chronic Spontaneous Urticaria.

Lakin E, Church M, Maurer M, Schmetzer O Theranostics. 2019; 9(3):829-836.

PMID: 30809311 PMC: 6376472. DOI: 10.7150/thno.29902.

High-density IgE recognition of the major grass pollen allergen Phl p 1 revealed with single-chain IgE antibody fragments obtained by combinatorial cloning.

Madritsch C, Gadermaier E, Roder U, Lupinek C, Valenta R, Flicker S J Immunol. 2015; 194(5):2069-78.

PMID: 25637023 PMC: 4476238. DOI: 10.4049/jimmunol.1402185.

Multiple independent IgE epitopes on the highly allergenic grass pollen allergen Phl p 5.

Levin M, Rotthus S, Wendel S, Najafi N, Kallstrom E, Focke-Tejkl M Clin Exp Allergy. 2014; 44(11):1409-19.

PMID: 25262820 PMC: 4278554. DOI: 10.1111/cea.12423.

The human IgE repertoire.

Gadermaier E, Levin M, Flicker S, Ohlin M Int Arch Allergy Immunol. 2013; 163(2):77-91.

PMID: 24296690 PMC: 4497803. DOI: 10.1159/000355947.

References

Andreasson U, Flicker S, Lindstedt M, Valenta R, Greiff L, Korsgren M . The human IgE-encoding transcriptome to assess antibody repertoires and repertoire evolution. J Mol Biol. 2006; 362(2):212-27. DOI: 10.1016/j.jmb.2006.06.062. View

Dolecek C, Vrtala S, Laffer S, Steinberger P, Kraft D, Scheiner O . Molecular characterization of Phl p II, a major timothy grass (Phleum pratense) pollen allergen. FEBS Lett. 1993; 335(3):299-304. DOI: 10.1016/0014-5793(93)80406-k. View

Christensen L, Holm J, Lund G, Riise E, Lund K . Several distinct properties of the IgE repertoire determine effector cell degranulation in response to allergen challenge. J Allergy Clin Immunol. 2008; 122(2):298-304. DOI: 10.1016/j.jaci.2008.05.026. View

Lefranc M . Nomenclature of the human immunoglobulin heavy (IGH) genes. Exp Clin Immunogenet. 2001; 18(2):100-16. DOI: 10.1159/000049189. View

Padavattan S, Flicker S, Schirmer T, Madritsch C, Randow S, Reese G . High-affinity IgE recognition of a conformational epitope of the major respiratory allergen Phl p 2 as revealed by X-ray crystallography. J Immunol. 2009; 182(4):2141-51. DOI: 10.4049/jimmunol.0803018. View

Bischoff S . Role of mast cells in allergic and non-allergic immune responses: comparison of human and murine data. Nat Rev Immunol. 2007; 7(2):93-104. DOI: 10.1038/nri2018. View

Horst A, Hunzelmann N, Arce S, Herber M, Manz R, Radbruch A . Detection and characterization of plasma cells in peripheral blood: correlation of IgE+ plasma cell frequency with IgE serum titre. Clin Exp Immunol. 2002; 130(3):370-8. PMC: 1906552. DOI: 10.1046/j.1365-2249.2002.02025.x. View

Vrtala S, Sperr W, Reimitzer I, Van Ree R, Laffer S, Muller W . cDNA cloning of a major allergen from timothy grass (Phleum pratense) pollen; characterization of the recombinant Phl pV allergen. J Immunol. 1993; 151(9):4773-81. View

Eckl-Dorna J, Pree I, Reisinger J, Marth K, Chen K, Vrtala S . The majority of allergen-specific IgE in the blood of allergic patients does not originate from blood-derived B cells or plasma cells. Clin Exp Allergy. 2012; 42(9):1347-55. DOI: 10.1111/j.1365-2222.2012.04030.x. View

10.

Vrtala S, Grote M, Duchene M, Van Ree R, Kraft D, Scheiner O . Properties of tree and grass pollen allergens: reinvestigation of the linkage between solubility and allergenicity. Int Arch Allergy Immunol. 1993; 102(2):160-9. DOI: 10.1159/000236567. View

11.

van Neerven R, Knol E, Ejrnaes A, Wurtzen P . IgE-mediated allergen presentation and blocking antibodies: regulation of T-cell activation in allergy. Int Arch Allergy Immunol. 2006; 141(2):119-29. DOI: 10.1159/000094714. View

12.

Edwards M, Brouwer W, Choi C, Ruhno J, Ward R, Collins A . Analysis of IgE antibodies from a patient with atopic dermatitis: biased V gene usage and evidence for polyreactive IgE heavy chain complementarity-determining region 3. J Immunol. 2002; 168(12):6305-13. DOI: 10.4049/jimmunol.168.12.6305. View

13.

Lefranc M . Nomenclature of the human immunoglobulin kappa (IGK) genes. Exp Clin Immunogenet. 2001; 18(3):161-74. DOI: 10.1159/000049195. View

14.

Jakobsen C, Bodtger U, Kristensen P, Poulsen L, Roggen E . Isolation of high-affinity human IgE and IgG antibodies recognising Bet v 1 and Humicola lanuginosa lipase from combinatorial phage libraries. Mol Immunol. 2004; 41(10):941-53. DOI: 10.1016/j.molimm.2004.05.009. View

15.

Collins A, Sewell W, Edwards M . Immunoglobulin gene rearrangement, repertoire diversity, and the allergic response. Pharmacol Ther. 2003; 100(2):157-70. DOI: 10.1016/j.pharmthera.2003.07.002. View

16.

LARKIN M, Blackshields G, Brown N, Chenna R, McGettigan P, McWilliam H . Clustal W and Clustal X version 2.0. Bioinformatics. 2007; 23(21):2947-8. DOI: 10.1093/bioinformatics/btm404. View

17.

Geha R, Jabara H, Brodeur S . The regulation of immunoglobulin E class-switch recombination. Nat Rev Immunol. 2003; 3(9):721-32. DOI: 10.1038/nri1181. View

18.

Flicker S, Steinberger P, Norderhaug L, Sperr W, Majlesi Y, Valent P . Conversion of grass pollen allergen-specific human IgE into a protective IgG(1) antibody. Eur J Immunol. 2002; 32(8):2156-62. DOI: 10.1002/1521-4141(200208)32:8<2156::AID-IMMU2156>3.0.CO;2-A. View

19.

Flicker S, Vrtala S, Steinberger P, Vangelista L, Bufe A, Petersen A . A human monoclonal IgE antibody defines a highly allergenic fragment of the major timothy grass pollen allergen, Phl p 5: molecular, immunological, and structural characterization of the epitope-containing domain. J Immunol. 2000; 165(7):3849-59. DOI: 10.4049/jimmunol.165.7.3849. View

20.

Gadermaier E, Flicker S, Aberer W, Egger C, Reider N, Focke M . Analysis of the antibody responses induced by subcutaneous injection immunotherapy with birch and Fagales pollen extracts adsorbed onto aluminum hydroxide. Int Arch Allergy Immunol. 2009; 151(1):17-27. DOI: 10.1159/000232567. View