| OBJECTIVEThis study is designed to investigate the expression and clinical significance of PD-L1 in peripheral blood and synovial fluid CD15~+CD3~-neutrophils in patients with rheumatoid arthritis(RA),and to elucidate the correlation between the percentage of PD-L1 positive neutrophils and ESR,CRP,autoantibodies and RA disease activity.METHODSThis study collected 88 patients with RA who met the inclusion criteria from the Department of Rheumatology and Immunology,the First Affiliated Hospital of Nanchang University,and 52 healthy controls(HC)who met the inclusion criteria from the health check-up center.The peripheral blood of all subjects and the synovial fluid of 27 RA patients in the RA group were collected.The medical history,physical examination,clinical manifestations and laboratory results of all RA patients were recorded.The expression levels of PD-1,PD-L1,Tim-3,CD40,TIGIT,CD80 and CD86 on CD15~+CD3~-neutrophil in peripheral blood and synovial fluid of all research objects were detected by flow cytometry.The relationship between the number and proportion of CD15~+CD3~-neutrophils expressing PD-L1 in RA patients and their autoimmune responses,ESR,CRP and disease activity was further analyzed.RESULTS1.The percentage of PD-L1 positive neutrophils in RA disease group was(21.05±2.206)%,which was significantly higher than HC group(5.565±1.039)%.The difference was statistically significant(P<0.0001).There was no significant difference in the percentage of PD1,Tim-3,TIGIT,CD40,CD80 or CD86-positive neutrophils between the RA disease group and the HC group.2.The percentage of PD-L1 positive neutrophils in the synovial fluid of the RA disease group was(49.29±5.502)%,which was significantly higher than the peripheral blood of the HC group(5.565±1.039)%.The difference was statistically significant(P<0.0001).At the same time,the percentage of PD-L1 positive neutrophils in the synovial fluid of the RA disease group was significantly higher than the peripheral blood of the RA disease group(17.47±2.318)%,and the difference was statistically significant(P<0.0001).3.Correlation analysis between the percentage of PD-L1 positive neutrophils and auto-antibodies in RA patients:In RA group,the percentage of PD-L1 positive neutrophils in the peripheral blood of RF-positive patients was(35.15±3.442)%,which is higher than the corresponding negative group(15.55±3.561)%.The difference was statistically significant(P=0.0105).The percentage of PD-L1-positive neutrophils in peripheral blood of anti-CCP antibody positive patients was(33.69±3.804)%,which was higher than that of the corresponding negative group(24.39±4.104)%,but the difference did not reach statistical significance(P=0.197).The percentage of PD-L1 positive neutrophils in joint synovial fluid was not significantly correlated with auto-antibodies(P>0.05).4.Correlation analysis of PD-L1 positive neutrophils percentage with ESR and CRP in RA patients:The percentage of PD-L1 positive neutrophil was positively correlated with ESR in the RA group(r~2=0.1105,P=0.0068)and positively correlated with CRP(r~2=0.1183,P=0.0155).The percentage of PD-L1 positive neutrophils in joint synovial fluid was not correlated with ESR and CRP(P>0.05).5.Correlation analysis between PD-L1 positive neutrophil percentage and DAS28score in RA patients:In the RA group,the percentage of PD-L1 positive neutrophils in peripheral blood of patients with DAS28≥2.6 was(20.62±2.956)%,which is higher than the corresponding negative group(9.516±2.636)%.The difference was statistically significant(P=0.029).The percentage of PD-L1 positive neutrophils was positively correlated with the DAS28 score(r~2=0.067,P=0.0341).There was no significant correlation between the percentage of PD-L1 positive neutrophils in the synovial fluid of RA patients and the DAS28 score(P>0.05).CONCLUSIONThe expression level of PD-L1 on neutrophil is elevated in RA patients,and the proportion of PD-L1 positive neutrophils is related with ESR,CRP,auto-antibodies,and RA disease activity. |
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