IgE production in CD40/CD40L cross-talk of B and mast cells and mediator release via TGase 2 in mouse allergic asthma

TGase 2 is over-expressed in a variety of inflammatory diseases including allergic asthma. This study aimed to investigate the role of TGase 2 on IgE production and signaling pathways in mast cell activation related to OVA-induced allergic asthma. Bone marrow-derived mast cells (BMMCs) isolated from...

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Bibliographic Details
Published in:Cellular signalling 2013-06, Vol.25 (6), p.1514-1525
Main Authors: Hong, Gwan Ui, Park, Bum Soo, Park, Jung Won, Kim, Soo Youl, Ro, Jai Youl
Format: Article
Language:English
Subjects:
Allergic asthma
Animals
Asthma - immunology
Asthma - metabolism
Asthma - pathology
B cells
B-Lymphocytes - cytology
B-Lymphocytes - metabolism
Bone Marrow Cells - cytology
Bone Marrow Cells - metabolism
Calcium - metabolism
CD40 Antigens - metabolism
CD40 Ligand - metabolism
Cell surface molecules
Coculture Techniques
Cytokines - genetics
Cytokines - metabolism
Female
GTP-Binding Proteins - antagonists & inhibitors
GTP-Binding Proteins - genetics
GTP-Binding Proteins - metabolism
IgE
Immunoglobulin E - metabolism
Lung - metabolism
Mast cells
Mast Cells - cytology
Mast Cells - metabolism
Mast Cells - pathology
Matrix Metalloproteinases - genetics
Matrix Metalloproteinases - metabolism
Mice
Mice, Inbred C57BL
Mice, Knockout
RNA Interference
RNA, Small Interfering - metabolism
Signal Transduction
Tissue Inhibitor of Metalloproteinases - genetics
Tissue Inhibitor of Metalloproteinases - metabolism
Transglutaminase 2 (TGase 2)
Transglutaminases - antagonists & inhibitors
Transglutaminases - genetics
Transglutaminases - metabolism
Tumor Necrosis Factor-alpha - metabolism
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Summary:TGase 2 is over-expressed in a variety of inflammatory diseases including allergic asthma. This study aimed to investigate the role of TGase 2 on IgE production and signaling pathways in mast cell activation related to OVA-induced allergic asthma. Bone marrow-derived mast cells (BMMCs) isolated from WT or TGase 2−/− mice were activated with Ag/Ab (refer to act-WT-BMMCs and act-KO-BMMCs, respectively). B cells isolated from splenocytes were activated with anti-mouse IgM (act-B cells), and B cells were co-cultured with BMMCs. WT and TGase 2−/− mice were sensitized and challenged with OVA adsorbed in alum hydroxide. Intracellular Ca2+ ([Ca2+]i) levels were determined by fluorescence intensity; IgE, mediators and TGase 2 activity by ELISA; the CD138 expression by FACS analyzer; cell surface markers and signal molecules by Western blot; NF-κB by EMSA; co-localization of mast cells and B cells by immunohistochemistry; Fcε RI-mediated mast cell activation by PCA test; expression of cytokines, MMPs, TIMPs, TLR2 and FcɛRI by RT-PCR. In vitro, act-KO-BMMCs reduced the [Ca2+]i levels, NF-κB activity, expression of CD40/CD40L, plasma cells, total IgE levels and TGase 2 activity in act-B cells co-cultured with act-BMMCs, expression of inflammatory cytokines and MMPs2/9, release of mediators (TNF-α, LTs and cytokines), and activities of signal molecules (PKCs, MAP kinases, I-κB and PLA2), which were all increased in act-WT-BMMCs. TGase 2 siRNA transfected/activated-BMMCs reduced all responses as same as those in act-KO-BMMCs. In allergic asthma model, TGase 2−/− mice protected against PCA reaction, OVA-specific IgE production and AHR, and they reduced co-localization of mast cells and B cells or IgE in lung tissues, expression and co-localization of surface molecules in mast cells (c-kit and CD40L) and B cells (CD23 and CD40), inflammatory cells including mast cells, goblet cells, amounts of collagen and mediator release in BAL fluid and/or lung tissues, which were all increased in WT mice. TLR expression in TGase 2−/− mice did not differ from those in WT mice. Our data suggest that TGase 2 expression and Ca2+ influx required by bidirectional events in mast cell activation facilitate IgE production in B cells via up-regulating mast cell CD40L expression, and induce the expression of numerous signaling molecules associated with airway inflammation and remodeling in allergic asthma. •We first found that TGase 2 is expressed in mast cells.•TGase 2 expressed in mast cells e
ISSN:0898-6568
1873-3913
DOI:10.1016/j.cellsig.2013.03.010