Local Group 2 Innate Lymphoid Cell Affect The Manifestations And Inflammatory Cell Changes Of Fungal Keratitis In Mice

BackgroundFungal keratitis is a common cause of corneal lesions caused by fungal infections in our country,the blinding rate is extremely high,and mainly occurs in rural areas of young workers.Fungal meningitis is the main cause of corneal disease caused by plant trauma,in the previous epidemiological survey found that in our country the main pathogenic Fusarium and Aspergillus.In the treatment of fungal keratitis autoimmune state determines the fungal infection and prognosis,in the past study,the immune cells are mainly neutrophils,macrophages,monocytes and NK cells。In recent years,the discovery of a new immune cell may provide a new idea and direction for the participation and treatment of fungal keratitis-associated immune cells,that is ILCs.It has been proved in the early stage that at least ILC2 s exist in the cornea,so we hope to observe the changes of the disease course of fungal keratitis through the early intervention of ILC2 s,the influence of the inflammatory cells in different periods and the influence on the structure of the cornea in the later period.The experiment further in-depth understanding of the pathogenesis of fungal keratitis,provide new ideas for the treatment of fungal keratitis.ObjectiveTo investigate the effects of ILC2 s on the repair of fungal keratitis and the changes of inflammatory cells.Materials and methodsEighty-four male C57BL/6J mice of SPF grade were used in the experiment.All mice were 8-10 weeks of age and weighed 24-28 g.Forty-two of these mice were subconjunctivally injected with Anti-CD90.2 to remove ILC2 s.The mice were divided into Anti-CD90.2 group and WT group.,and the fungal keratitis model was prepared for each group of mice.The corneas were scored and photographed at 12 h,24h,48 h,72h,96 h,120h and 168 h respectively.The mice were randomly divided into 12 h,24h,48 h,72h,120 h and 168 h Group 8 eyes(n = 8).At corresponding time points to be put to death in mice and materials,to spread the whole piece of cornea with anti-mouseF4 PE / 80 respectively,FITC Rat anti-mouseGr-1 mark macrophages and neutrophils,applied research fluorescence microscope photograph spread sheet,calculate the area of neutrophils and macrophages.In addition,4 eyes(n=4)of each subgroup at 24 h,48 h,72 h,96 h,120 h,and 168 h were collected from the eyeballs and placed in a fixed solution to make pathological sections and HE staining.Changes in corneal structure at the time point.Statistical data were determined and analyzed using SPSS24.0 software.Results1.Compared with the clinical scores of Anti-CD90.2 group and WT group at the same time point,there was no statistically significant difference in the clinical scores of fungal infection of 12 h,24h,36 h,and 48 h,respectively(P=0.626,0.389,0.306,0.106>0.05).The clinical scores of fungal infection of 72 h,120h and 168 h corneal lesions were statistically significant(P=0.025,0.014,0.005<0.05).The clinical score of Anti-CD90.2 group of corneal lesions was significantly lower than that of WT group,and the corneal transparency was also better than that of WT group after the fungal infection of 48 h.2.Both groups of neutrophils increased early,36 h corneal neutrophil infiltration reached its highest point,and then began to decline.Compared with the same time point of neutrophil infiltration in Anti-CD90.2 group and WT group,the difference was statistically significant at 12 h,24h,36 h and 168h(P = 0.000);72h and 120 h were also statistically significant(P = 0.002,0.017 <0.05).3.In WT group,the infiltration amount of macrophages gradually increased with the time of fungal infection,after 72 h gradually decreased.However,the Anti-CD90.2 group showed no significant changes in the infiltration amount of macrophages in the early stage of fungal infection,and there was no statistically significant difference between the fungal infection of 36 h,24h and 12h(P =0.080,0.946>0.05).There was no significant difference in the time points between the two groupsbetween the two groups of the Anti-CD90.2 group and the WT group of macrophage corneal infiltration at the same time point.(P >0.05)The comparison of the time points between the two groups in the fungal infection of 48 h,72h,120 h and 168 h was statistically significant(P =0.000).Conclusions1.ILC2 s increase the severity and perforation rate of fungal keratitis.2.ILC2 s enhance neutrophil and macrophage infiltration of the lesion area.