| Objectives: The aim of this research is to study IIDDs in three aspects: diagnosis,cohort establishment,and treatment.AQP4-Ig G,MOG-Ig G,and GFAP-Ig G antibody detection are essential to the diagnosis of these autoimmune diseases.Significantly,the laboratory-testing platform is the basis for further study.In addition,establishment of cohort facilitates follow-up and further management of patients.The analysis of antigen-specific antibody secreting cells and important pathogenic factors in patients can contribute to guide treatment and provide potential targets for treatment.Methods: We recruited 109 IIDDs patients hospitalized in the department of Neurology and Ophthalmology of First Affiliated Hospital of Fujian Medical University from March 2017 to October 2019.AQP4-Ig G,MOG-Ig G,and GFAP-Ig G were detected by CBA assay.We further analyzed clinical characteristics,laboratory tests,and MRI data of these patients.PBMCs of NMOSD patients who were positive for AQP4-IgG antibody were cultured in vitro and added IL-2 and R848 cytokines without exogenous antigens.Then we analyzed the proportion of CD19+CD38+CD27+ cells by flow cytometry.The cell culture supernatant was collected and tested specific antibody by CBA.BAFF,a key factor in NMOSD pathogenesis,may be a potential therapeutic target.In this study,we investigated the potential regulators of BAFF through whole transcriptome sequencing,dual-luciferase reporter gene assay,magnetic bead sorting,and quantitative real-time PCR to confirm the molecular regulatory mechanism of BAFF and seek potential diagnostic markers and therapeutic targets.Results:1.Among 109 IIDDs patients,63(57.8%)were serumal AQP4-Ig G positive,12(11.0%)were serumal MOG-Ig G positive,26(23.8%)were serumal GFAPα-Ig G positive,and 3 cases were double positive of AQP4-Ig G and MOG-Ig G.2.Summarize patients’ clinical symptoms,imaging features,antibody condition(Part 2).3.The necessity of detection of antibody subtype: 1)The AQP4-Ig G detection result of a patient with area postrema syndrome was suspicious,whose antibody subtype was AQP4-Ig G1 and AQP4-Ig M.2)A patient with MOG encephalomyelitis underwent a suspicious MOG-Ig G detection result while antibody subtype was MOG-Ig G1.4.Compared with healthy controls,PBMCs of AQP4-Ig G-positive patients showed a significant increase in ASCs after IL-2 and R848 factor added in vitro without AQP4 antigen stimulation.Subsequently,the supernatant of culture system was positive for AQP4-Ig G detected by CBA.5.The expression of MALAT1 and BAFF were significantly upregulated while mi R-30b-5p was significantly downregulated in monocytes of NMOSD patients,which demonstrated that BAFF expression was negatively correlated with mi R-30b-5p.Furthermore,MALAT1 is able to bind to and negatively modulate mi R-30b-5p.Conclusion: In this study,the first laboratory detection platform for antibody of neuroimmune diseases in Fujian Province was established.Detecting antibody and its subtype are essential for the diagnosis and treatment of IIDDs.When it comes to the therapeutic aspects,future therapeutic trends in NMOSD focus on specific antibody-secreting cells or key factors.The stimulation of ASCs from PBMCs without antigens suggests the presence of specific antibody-secreting cells in the patient’s peripheral blood,helping to guide treatment and provide the foundation for further experiments about ASCs.Additionally,it is an important means of studying specific B cells in autoantibody-mediated diseases.Furthermore,the study of BAFF which is a key factor in NMOSD may provide a potential therapeutic target for the disease.Last but not the least,the series of study of IIDDs have expanded the understanding of the disease in both diagnostic and therapeutic terms,and the establishment of the platform and various experimental methods can serve as a cornerstone for subsequent further studies. |
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