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Filling the Antibody Pipeline in Allergy: PIPE Cloning of IgE, IgG1 and IgG4 against the Major Birch Pollen Allergen Bet v 1

Authors :
Köhler, Verena K.
Crescioli, Silvia
Fazekas-Singer, Judit
Bax, Heather J.
Hofer, Gerhard
Pranger, Christina L.
Hufnagl, Karin
Bianchini, Rodolfo
Flicker, Sabine
Keller, Walter
Karagiannis, Sophia N.
Jensen-Jarolim, Erika
Source :
International Journal of Molecular Sciences, Vol 21, Iss 5693, p 5693 (2020), International Journal of Molecular Sciences, Volume 21, Issue 16
Publication Year :
2020
Publisher :
MDPI AG, 2020.

Abstract

Birch pollen allergy is among the most prevalent pollen allergies in Northern and Central Europe. This IgE-mediated disease can be treated with allergen immunotherapy (AIT), which typically gives rise to IgG antibodies inducing tolerance. Although the main mechanisms of allergen immunotherapy (AIT) are known, questions regarding possible Fc-mediated effects of IgG antibodies remain unanswered. This can mainly be attributed to the unavailability of appropriate tools, i.e., well-characterised recombinant antibodies (rAbs). We hereby aimed at providing human rAbs of several classes for mechanistic studies and as possible candidates for passive immunotherapy. We engineered IgE, IgG1, and IgG4 sharing the same variable region against the major birch pollen allergen Bet v 1 using Polymerase Incomplete Primer Extension (PIPE) cloning. We tested IgE functionality and IgG blocking capabilities using appropriate model cell lines. In vitro studies showed IgE engagement with Fc&epsilon<br />RI and CD23 and Bet v 1-dependent degranulation. Overall, we hereby present fully functional, human IgE, IgG1, and IgG4 sharing the same variable region against Bet v 1 and showcase possible applications in first mechanistic studies. Furthermore, our IgG antibodies might be useful candidates for passive immunotherapy of birch pollen allergy.

Details

Language :
English
ISSN :
16616596 and 14220067
Volume :
21
Issue :
5693
Database :
OpenAIRE
Journal :
International Journal of Molecular Sciences
Accession number :
edsair.dedup.wf.001..5611d77788a9dc004b38669842bb25f5