A Preliminary Study On The Immune Repertoire Of Peripheral Blood IgH-CDR3 In Patients With Nmosd

Background Neuromyelitis optica spectrum disorders(NMOSD)is an autoimmune demyelinating disease of the central nervous system with severe disability,which is characterized by severe opticneuritis and / or transverse myelitis.Its exact etiology and pathogenesis are still not been revealed.In order to explore more effective methods for diagnosis,prevention and treatment,domestic and foreign researchers have made extensive exploration,aiming at reducing the disability rate of the disease and improving its prognosis,so as to improve patien ts' living quality witch with NMOSD.Objective An Immune Repertoire of Ig H-CDR3 in peripheral blood of patients with NMOSD was established.To explore the correlation between the changes in the diversity of Immune Repertoire and the dominant use of the Ig H-CDR3 sequence and the occurrence,progression and therapeutic intervention of NM OSD.The characteristics of Immune Repertoire and the diversity of the immune system of NMOSD patients were analyzed.Materials and method Fresh peripheral blood of female patients with NMOSD which were diagnosed in the hospital from May 2015 to December 2017 was collected.Clinical data and laboratory examination data were collected,including 6 cases of NMOSD patients,which were divided into early stage and stable period,and 6 cases of female healthy volunteers who underwent physical examination in our hospital during the same period as the control group.A total of 18 samples were included.All the peripheral blood samples of the subjects were sent to Wu Han BGI for testing.The nucleotide sequence of the IgH-CDR3 region of B cells in peripheral blood was specifically amplified by multiple PCR,and the second generation of high-throughput sequencing technology was used to sequence the immunome library.The sequencing results and clinical data were combined to analyze the Immune Repertoire characteristics of NMOSD patients.Results A total of 18 samples of peripheral blood B cells IgH-CDR3 were successfully constructed included 6 patients with NMOSD and 6 healthy volunteers.It was found that the total number and types of IgH-CDR3 in peripheral blood of NMOSD patients decreased compared with that of healthy volunteers.Compared with healthy volunteers,the diversity of Ig H-CDR3 sequence in the N MOSD patients group was also lower,and the diversity of IgH-CDR3 sequence in the early NMOSD patients was generally higher than that in the stable patients.Compared with the healthy volunteers group,it was revealed that the use f requency of alleles IGHV1-69,IGHV3-23,IGHJ1,IGHJ2,IGHJ3 and IGHJ4 in NMOSD patients was higher than that in the normal control group,in contrast,the use frequency of alleles IGHV1-8,IGHV3-30,IGHJ5 and IGHJ6 was lower than that of the normal control group.In this study,two disease characteristic sequences were selected: “CASSICLGSGCGGYYYGMDVW”?”CARPNYYGS VRTGYGMDVW”.The characteristic sequence "CASSICLGSGCGGYYYGMDV W" was highly expressed in the early NMOSD treatment group,and its clonal frequency was significantly higher than that of the stable NMOSD patients and healthy volunteers,and the difference was statistically significant.Conclusion This study established the Immune Repertoire of Ig H-CDR3 in the peripheral blood of 6 patients with NMOSD,and the patients were divided in to early stage and stable stage.By analyzing the diversity of the IgH-CDR3 of peripheral blood B cells in NMOSD patients,the expression of disease charact eristic advantage extraction sequence and the use frequency of V and J genes,and then combined with the clinical data of NMOSD patients,we found that the clonal amplification of disease characteristic advantage extraction sequence and the diversity of Immune Repertoire could effectively reflect the disease status of NMOSD patients and evaluate the therapeutic effect.The study also revealed that the occurrence and progression of NMOSD may be closely related to the mass cloning and amplification of peripheral blood B cells with specific Ig HCDR3 sequence.Further research is needed to provide relevant molecular targets and important theoretical basis for clinical immunotherapy.