Differential Expression And Significance Of Cytokine In Anti-N-methyl-D-aspartate Receptor Encephalitis

Objective:Anti-N-Methyl-D-Aspartate(NMDA)receptor encephalitis is an autoimmune encephalitis mediated by anti-NMDA receptor antibody,and its diagnosis mainly depends on the detection of NMDA receptor antibody in cerebrospinal fluid.However,the pathogenesis of this disease is not completely clear at present,and some studies have shown that antibody titer is not significantly correlated with the severity and prognosis of this disease,suggesting that other factors may be involved in the occurrence and development of anti-NMDA receptor encephalitis.Therefore,it is of great significance to find biomarkers related to the occurrence,development and prognosis of anti-NMDA receptor encephalitis for further exploration of the its pathogenesis.Method:1.In the first part of this study,6 patients with anti-NMDA receptor encephalitis and 6 patients with non-inflammatory neurological diseases were included.Cerebrospinal fluid samples were obtained.Human inflammatory cytokines arrays were used to screen the differentially expressed cytokines in cerebrospinal fluid samples of two groups.Functional enrichment and hierarchical clustering analysis of the differentially expressed proteins was also performed.2.The second part expanded the sample size which including 13 patients with anti-NMDA receptor encephalitis and 13 patients with non-inflammatory neurological diseases.Cerebrospinal fluid samples and clinical data were collected.The differentially expressed cytokines were verified.At the same time,the expression of some other biomarkers in the cerebrospinal fluid specimens of patients with anti-NMDA receptor encephalitis and control group were detected and compared by ELISA.3.The correlations between candidate biomarkers and the clinical characteristics of patients with anti-NMDA receptor encephalitis were analysed,and the optimal diagnostic efficiency thresholds of candidate biomarkers were evaluated by ROC curve.Results:1.Three differentially expressed proteins were screened from the cerebrospinal fluid of patients with anti-NMDA receptor encephalitis by protein chip technology,in which the expressions of c IAP-1 and MCSF were up-regulated and the expression of LAMP-1 was down-regulated.2.Go pathway analysis shows that,in terms of biological process,differentially expressed proteins are mainly involved in regulating macrophage colony-stimulating factor response,regulating cytokine response and regulating the proliferation and migration of microglia;in terms of cell composition,differentially expressed proteins are mainly located in phagolysosome,cytolytic granule,autolysosome,CD40 receptor complexe,secondary lysosome,synaptic vesicle membrane and other parts;in terms of molecular function,they mainly mediate apoptosis,exogenous protein binding,ubiquitin binding,ubiquitin-like protein binding,and so on.KEGG pathway analysis presented that the differentially expressed proteins were mainly concentrated in the TNF signaling pathway.3.ELISA was used to test the level of screened differentially expressed proteins in cerebrospinal fluid of patients with anti-NMDA receptor encephalitis,the levels of some cytokines reported previously in cerebrospinal fluid of anti-NMDA receptor encephalitis group and control group were detected simultaneously.The results showed that compared with the control group,the levels of c IAP-1,MCSF,LAMP-1,CXCL13 and NLRP3 increased significantly.Among them,the difference of LAMP-1between groups was opposite to that detected by protein chip technology.4.The level of CXCL13 was positively correlated with the duration of antiNMDA receptor encephalitis,the highest m RS in the acute phase,and the m RS score at 3 months follow-up and NEOS score.The level of MCSF was positively correlated with the duration of the disease.The level of NLRP3 was significantly correlated with the content of cerebrospinal fluid protein.However,there was no significant correlation between the levels of all cytokines and gender,age,proinfection,cerebrospinal fluid leukocyte count,antibody titer,brain MRI and EEG abnormalities.5.The c IAP-1,MCSF,CXCL13 and NLRP3 levels in cerebrospinal fluid can be used as evaluation indicators of anti-NMDA receptor encephalitis.The AUC of c IAP-1 factor is closer to 1.Conclusion:The levels of c IAP-1,MCSF,CXCL13 and NLRP3 in cerebrospinal fluid can be used as potential biomarkers for evaluating the condition of anti-NMDA receptor encephalitis.These cytokines may be involved in the pathogenesis of anti-NMDA receptor encephalitis through biological processes such as neuroinflammation,microglia polarization,autophagy,and regulation of TNF signaling pathway.