Etude du TLR9 à la membrane plasmique des lymphocytes B : caractérisation des anticorps anti-TLR9 commerciaux

Abstract : Regulatory B cells (Breg) play a key role in regulating the immunological response.This regulatory behaviour can be induced by stimulation of the B cells. Toll-like receptor 9 (TLR9), an innate immune receptor, participate to that induction. TLR9 signalling pathway is deficient in B cells from systemic lupus erythematosus patients suffering from chronic hyper-inflammation and having defective Breg cells. To study TLR9, commercial antibodies are commonly used. Among them, the anti-TLR9 monoclonal antibody 26C593.2 targets an epitope located at the plasma membrane of the B cells. Moreover, this particular antibody activates the B cells in vitro and blocks the CpGODN effects, an agonist of the endosomal TLR9. In this work, we show that five commercial antibodies are not specific for the TLR9 protein and recognize other antigens.However, four tested antibodies can recognize recombinant TLR9 molecule. In order to avoid the specificity problem, we achieved to build a chimeric TLR9 protein expressing a hemagglutinin tag at its C-terminal region (TLR9-HA). Three antibodies efficiently target this overexpressed protein in transfected cells.However, no TLR9-HA molecule seems to be located at the plasma membrane of transfected cells. This approach allowed us to demonstrate that anti-TLR9 antibodies recognize another target than TLR9. It would of interest to identify that protein in order to better understand the mouse monoclonal anti-TLR9 antibody effect on B cell activation and Breg properties.
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Submitted on : Wednesday, December 5, 2018 - 11:37:54 AM
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Marie Cumin. Etude du TLR9 à la membrane plasmique des lymphocytes B : caractérisation des anticorps anti-TLR9 commerciaux. Immunologie. Université de Bretagne occidentale - Brest, 2018. Français. ⟨NNT : 2018BRES0042⟩. ⟨tel-01945286v2⟩

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