AIM： To investigate how macrophage inducible C-type lectin（Mincle） influences inflammation in mice fungal keratitis induced by Aspergillus fumigatus（A. fumigatus）.METHODS： C57 BL/6 mice were infected with A. fum...AIM： To investigate how macrophage inducible C-type lectin（Mincle） influences inflammation in mice fungal keratitis induced by Aspergillus fumigatus（A. fumigatus）.METHODS： C57 BL/6 mice were infected with A. fumigatus after pretreated with Mincle agonist TDB or Mincle neutralizing antibody（MincleAb）, taking DMSO or IgG as control group respectively. The cornea lesions were monitored with slit-lamp microscope and evaluated by clinical score. Mincle expression was assessed using reverse transcriptionploymerase chain reaction（RT-PCR） and immunostaining. The expression of cytokines（IL-1β, TNF-α and IL-6） chemokines（CXCL-1 and MIP-2） was determined by RTPCR and ELISA. Neutrophil infiltration was observed by immunostaining. The levels of nitric oxide（NO） generated by corneas were tested by Griess reaction. RESULTS： Mincle mRNA and protein levels were higher in infected corneas than normal corneas of C57 BL/6 mice, saving clinical scores revealed differences. When pretreated with Mincle agonist TDB, the mRNA and protein levels of IL-1β, TNF-α and IL-6 in infected corneas were significantly increased compared with the control group（P〈0.01）. Results of the counterpart in corneas pretreated with Mincle neutralizing antibody was decreased consistently（P〈0.01）. Expression of CXCL1 and MIP-2 mRNA levels were upregulated in TDB group and down-regulated in Mincle Ab group（P〈0.01）, coincide with neutrophil aggregation degree in corneas showed by immunostaining. As for the concentration of NO, it was promoted in TDB group compared with DMSO control group, and decreased in Mincle Ab group compared with Ig G control group.CONCLUSION： Mincle plays a dual role in mice fungal keratitis. It participates in the innate immune system by enhancing inflammation. What＇s more, Mincle can mediate cytotoxic effects by regulating the formation of NO.展开更多
基金Supported by the National Natural Science Foundation of China(No.81470609No.81500695)
文摘AIM： To investigate how macrophage inducible C-type lectin（Mincle） influences inflammation in mice fungal keratitis induced by Aspergillus fumigatus（A. fumigatus）.METHODS： C57 BL/6 mice were infected with A. fumigatus after pretreated with Mincle agonist TDB or Mincle neutralizing antibody（MincleAb）, taking DMSO or IgG as control group respectively. The cornea lesions were monitored with slit-lamp microscope and evaluated by clinical score. Mincle expression was assessed using reverse transcriptionploymerase chain reaction（RT-PCR） and immunostaining. The expression of cytokines（IL-1β, TNF-α and IL-6） chemokines（CXCL-1 and MIP-2） was determined by RTPCR and ELISA. Neutrophil infiltration was observed by immunostaining. The levels of nitric oxide（NO） generated by corneas were tested by Griess reaction. RESULTS： Mincle mRNA and protein levels were higher in infected corneas than normal corneas of C57 BL/6 mice, saving clinical scores revealed differences. When pretreated with Mincle agonist TDB, the mRNA and protein levels of IL-1β, TNF-α and IL-6 in infected corneas were significantly increased compared with the control group（P〈0.01）. Results of the counterpart in corneas pretreated with Mincle neutralizing antibody was decreased consistently（P〈0.01）. Expression of CXCL1 and MIP-2 mRNA levels were upregulated in TDB group and down-regulated in Mincle Ab group（P〈0.01）, coincide with neutrophil aggregation degree in corneas showed by immunostaining. As for the concentration of NO, it was promoted in TDB group compared with DMSO control group, and decreased in Mincle Ab group compared with Ig G control group.CONCLUSION： Mincle plays a dual role in mice fungal keratitis. It participates in the innate immune system by enhancing inflammation. What＇s more, Mincle can mediate cytotoxic effects by regulating the formation of NO.
