Biomarkers Of Ocular Autoimmune Inflammation&Immunemodulation Effecets Of A Novel Peptide H-CN

Purpose:The percentage of autoimmune inflammatory diseases in ocular inflammatory diseases is increasing gradually.Among them,autoimmune uveitis and autoimmune optic neuritis take an imporant role due to the poor vision prognosis,and are the major causes of blindness in young people.In the current paper,we took a comprehensive study on the diagnosis,pathophysiological mechanisms and treatment of autoimmune uveitis and autoimmune optic neuritis through the fundus imaging technology,biological technology and experimental methods.Methods:(1)The characteristics of active lesions in multifocal choroiditis(MFC)were investigated with OCTA and other multimodal fundus imaging techniques.(2)Macular layer thicknesses and retinal blood flow density of optic neuromyelitis(NMO)patients were measured with OCT and OCTA,and the relationship between fundus flow density and blood-brain barrier biomarkers was investigated.(3)To explore novel immunological biomarkers of NMO,the cerebrospinal fluid and serum of NMO patients were tested through membrane-protein chip and cytometric beads array,and Th subsets were analyzed by flow cytometry.(4)A novel peptide targeting the above immunological biomarkers was screened with bioinformatics techniques,and the experimental autoimmune uveitis(EAU)model and experimental autoimmune encephalitis(EAE)model were established to verify the immunomodulatory effects of the peptide.Results:(1)OCTA could effectively distinguish the inflammatory lesions and secondary CNV lesions in MFC patients,of which the utility is superior to other fundus imaging techniques.(2)The macula layer thicknesses(especially the inner layers)of NMO patients were thinner than those of the normal control(P<0.01).Also,the macular and peripapillary blood flow density of NMO patients were obviously smaller than the normal group.Moreover,there was a significant correlation between the peripapillary flow density and the serum blood-brain barrier biomarkers(P<0.01).(3)T cell dysfunction,including increase of Th17subset and relative reduction of Treg subset,along with the abnormal levels of serum IL-17A,IL-6 and other related inflammatory cytokines,take an important part in the mechanism of NMO.(4)H-CN can effectively change the subsets of CD4~+T cells in both peripheral immune organs and effected organs of EAE and EAU models.It could reduce the percentage of Th1 and Th17 subsets(P<0.01),increase the percentage of Treg subsets(P<0.01),then reduce the levels of IFN-r,IL-17A,IL-2,IL-6and TNF-a,and increase the level of IL-10.Besides it could reduce the clinical scores,improve visual function and attenuated the ocular pathological damge in EAU and EAE mice.Conclusion:OCTA provides a new imaging biomarker for MFC and NMO.It can be used in the disease diagnosis,differentiation and the study of pathophysiological mechanism.It also provides a new method for assessment the blood-brain barrier injury.In addition,the abnormalities of Th17 and Treg axis took an important parts in the pathogenesis of NMO.Targetting this novel immunolgical biomarker,we screened out a novel immunomodulatory peptide H-CN,which could reduce the Th1,Th17,and other proinflammatory T cell subsets,as well as promote the proliferation of Treg subset.Therefore,H-CN plays an immune regulation effect in the autoimmune inflammatory diseases and could be a promising treatment approach in ocular autoimmune abnormalities.