Abstract:

Objective To analyze the immunological regulation mechanism of the progression of diabetes to diabetic retinopathy. We used Flow cytometry to compare the differences of immune cell profiles in peripheral blood of healthy donors （HD）， diabetic patients （DM）， non-proliferative diabetic retinopathy patients （NPDR） and proliferative diabetic retinopathy patients （PDR）. Methods Peripheral blood was collected from HD， DM， NPDR and PDR， and peripheral blood immune cells were isolated by density gradient centrifugation. The 13-color flow cytometry antibody was co-stained and detected by flow cytometry. Combined with Flowjo and its plug-in， the cells were analyzed by dimensionality reduction and clustering. Finally， Spearman correlation analysis and receiver operating characteristic （ROC） curve were used to evaluate the diagnostic value of peripheral blood immune cells in predicting the progression from DM to DR stage. Results Peripheral blood immune cells were divided into 10 groups. Among the innate immune cells， the number of mono-classical cells in DM and PDR was higher than that in healthy people （P < 0.05）. In adaptive immune cells， B cells in DM and NPDR were higher than those in HD group （P < 0.05）， while NKT and T cells were lower than those in HD group （P < 0.05）. Compared with healthy people， B cells in PDR were increased （P < 0.05）， but T cells were decreased （P < 0.05）. In addition， B cells were positively correlated with each stage. And in the PDR stage， ROC curve analysis showed that the efficiency of B cells in predicting the progression of DM to DR was greater than that of other immune cells. Conclusion Differences were noted between the three stages of peripheral blood immune cell profiles in HD， DM and DR. In addition， B cells were positively correlated with each stage， and had the potential to predict the diagnostic value of disease progression.

Key words: diabetes mellitus, diabetic retinopathy, immune cells, flow cytometry