Prevention of Intestinal Allergy in Mice by rflaA: Ova Is Associated with Enforced Antigen Processing and TLR5-Dependent IL-10 Secretion by mDC

被引：17

作者：

Schuelke, Stefan ^{[1
]}

Wolfheimer, Sonja ^{[1
]}

Gadermaier, Gabriele ^{[2
]}

Wangorsch, Andrea ^{[1
]}

Siebeneicher, Susanne ^{[3
]}

Briza, Peter

Spreitzer, Ingo ^{[4
]}

Schiller, Dirk ^{[1
]}

Loeschner, Bettina ^{[4
]}

Uematsu, Satoshi ^{[5
]}

Ryffel, Bernard ^{[6
]}

Akira, Shizuo ^{[5
]}

Waibler, Zoe ^{[7
]}

Vieths, Stefan ^{[1
]}

Toda, Masako ^{[3
]}

Scheurer, Stephan ^{[1
]}

机构：

[1] Paul Ehrlich Inst, Div Allergol, Langen, Hessen, Germany

[2] Salzburg Univ, Dept Mol Biol, Christian Doppler Lab Allergy Diag & Therapy, A-5020 Salzburg, Austria

[3] Paul Ehrlich Inst, Jr Res Grp Expt Allergy Models, Langen, Hessen, Germany

[4] Paul Ehrlich Inst, Div Microbiol, Langen, Hessen, Germany

[5] Osaka Univ, Microbial Dis Res Inst, Dept Host Def, Osaka, Osaka, Japan

[6] Univ Orleans, CNRS, Dept Mol & Expt Immunol & Neurogenet, Orleans, France

[7] Paul Ehrlich Inst, Jr Res Grp Novel Vaccinat Strategies & Early Immu, Langen, Hessen, Germany

来源：

PLOS ONE | 2014年 / 9卷 / 02期

关键词：

FUSION PROTEIN; BACTERIAL FLAGELLIN; ADJUVANT ACTIVITY; IMMUNE-RESPONSE; VACCINE; CELLS; EXPRESSION; STIMULATION; ACTIVATION; TOLERANCE;

D O I：

10.1371/journal.pone.0087822

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Conjugated vaccines consisting of flagellin and antigen activate TLR5 and induce strong innate and adaptive immune responses. Objective of the present study was to gain further insight into the mechanisms by which flagellin fusion proteins mediate their immune modulating effects. In a mouse model of Ova-induced intestinal allergy a fusion protein of flagellin and Ova (rflaA:Ova) was used for intranasal and intraperitoneal vaccination. Aggregation status of flaA, Ova and flaA: Ova were compared by light scattering, uptake of fluorescence labeled proteins into mDC was analyzed, processing was investigated by microsomal digestion experiments. Mechanism of DC-activation was investigated using proteasome and inflammasome inhibitors. Immune responses of wildtype, IL-10(-/-), TLR5(-/-) mDCs and Ova-transgenic T cells were investigated. Mucosal and i.p.-application of rflaA: Ova were able to prevent allergic sensitization, suppress disease-related symptoms, prevent body weight loss and reduction in food uptake. Intranasal vaccination resulted in strongest suppression of Ova-specific IgE production. These protective effects were associated with increased aggregation of rflaA: Ova and accompanied by tenfold higher uptake rates into mDC compared to the mixture of both proteins. Microsomal digestion showed that stimulation with rflaA: Ova resulted in faster degradation and the generation of different peptides compared to rOva. rflaA: Ova-mediated activation of mDC could be suppressed in a dose-dependent manner by the application of both inflammasome and proteasome inhibitors. Using TLR5(-/-) mDC the rflaA: Ova induced IL-10 secretion was shown to be TLR5 dependent. In co-cultures of IL-10(-/-) mDC with DO11.10 T cells the lack of rflaA: Ova-mediated IL-10 secretion resulted in enhanced levels of both TH2 (IL-4, IL-5) and TH1 (IL-2 and IFN-y) cytokines. In summary, mucosal vaccination with flaA: Ova showed strongest preventive effect. Stimulation with rflaA: Ova results in strong immune modulation mediated by enhanced uptake of the aggregated fusion protein, likely resulting in a different processing by DC as well as stronger TLR5 mediated cell activation.

引用

页数：12