TLR9 played a more important role than TLR2 in the combination of maltose-binding protein and BCG-induced Th1 activation.
Our previous study demonstrated that maltose-binding protein (MBP) combined with BCG induced synergistic mouse Th1 activation in vivo. Here, to explore the mechanism of MBP combined with BCG on Th1 activation, mouse purified CD4+T cells were stimulated with MBP and BCG in vitro. The results showed that MBP combined with BCG synergistically increased IFN-γ production, accompanied with the upregulation of TLR2/9 expressions, suggesting that TLR2/9 were involved in the combination-induced Th1 activation. Next, TLR2 antibodies and TLR9 inhibitor were used to further analyze the effects of TLRs in Th1 activation. Results showed TLR2 antibody partly decreased MBP combined with BCG-induced IFN-γ production, MyD88 expression and IκB phosphorylation, indicating that TLR2-mediated MyD88-dependent pathway was involved in the MBP combined with BCG-induced Th1 activation. Moreover, MBP combined with BCG-induced Th1 activation was completely abrogated by TLR9 inhibitor, suggesting that TLR9-mediated MyD88-dependent pathway played a more important role than TLR2 in the combination-induced Th1 activation. Further study showed that TLR9 inhibitor downregulated TLR2 expression, suggesting that TLR9 signaling regulated TLR2 activation to favor Th1 resonse induced by MBP combined with BCG. Collectively, we demonstrated for the first time that the cross-talk of TLR2 and TLR9 triggered Th1 activation collaboratively and our findings provided valuable information about designing more effective adjuvant for cancer therapy.
Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun, China.
Weihua
W
Ni
Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun, China.
Fang
F
Wang
Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun, China.
Guomu
G
Liu
Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun, China.
Nannan
N
Zhang
Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun, China.
Hongyan
H
Yuan
Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun, China.
Jing
J
Jie
Department of Immunology, Norman Bethune College of Medicine, Jilin University, Changchun, China. Electronic address: taiguixiang@163.com.
Guixiang
G
Tai
D000818:Animals; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D008213:Lymphocyte Activation / Q000276:immunology*; D058619:Maltose-Binding Proteins / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D009163:Mycobacterium bovis / Q000276:immunology; D016133:Polymerase Chain Reaction; D020239:Receptor Cross-Talk; D018417:Th1 Cells / Q000276:immunology*; D051195:Toll-Like Receptor 2 / Q000276:immunology*; D051217:Toll-Like Receptor 9 / Q000276:immunology*
D058619:Maltose-Binding Proteins; C493486:Tlr2 protein, mouse; C495370:Tlr9 protein, mouse; D051195:Toll-Like Receptor 2; D051217:Toll-Like Receptor 9
BCG; MBP; Maltose-binding protein; TLR2/9 cross-talk; Th1 activation