Expression Of Regulatory B Cell And IL-17 In Peripheral Blood Of Patients With Primary Sj?gren's Syndrome

Objective:Primary sj?gren's syndrome?pSS?is a chronic autoimmune disease with the character of highly lymphocytes infiltration within salivary glands involved hyperglobulinemia.Clinically,the most frequent symptoms are xerostomia and xerophthalmia,often accompanied with multiple organ system involvement such as pulmonary interstitial fibrosis,nervous system lesions,and renal tubular acidosis.The etiology and pathogenesis of pSS are not very clear.Studies have found that the onset of pSS is associated with genetic and environmental factors,a variety of inflammatory cells and their relevant cytokines.The role of B lymphocytes in the pathogenesis of autoimmune diseases has being paid more attention increasingly.Studies have showed that regulatory B cell and IL-17 are closely related to pSS.This study intend to investigate the expression of CD19+CD24hiCD38hi Bregs and IL-17 in pSS patients and analyze the correlation between their expression levels and clinical symptoms,laboratory indexes,labial gland pathological grading,disease activity of index in order to provide a new diagnostic method and a new way of thinking for the pSS treatment.Methods:1 Case selection: The 30 untreated pSS patients were selected from those hospitalized in Department of Rheumatology and Clinical Immunology of Hebei General Hospital from March 2016 to November 2016,as experimental group.All the patients were fulfilled with 2002 European classification criteria for sj?gren's syndrome,without other connective tissue diseases,malignant tumors,history of head and neck radiotherapy,AIDS,graft versus host disease,hepatitis B or C virus replication,use of acetylcholine and/or Glucocorticoid.20 cases of normal healthy individual were enrolled as control group.2 Data collection: The basic information and clinical data of experimental subjects were collected,including WBC,RBC,Plt,Hb,ANA,anti-ENA,RF,IgA,G,M,CRP,ESR,C3,C4.3 The expression of CD19⁺CD24^hiCD38^hiBregs was detected by flow cytometry?FCM?.Enzyme linked immunosorbent assay?ELISA?was used to determine the level of serum IL-17 in both groups.4 Labial salivary gland tissues were stained by HE and graded according to the degree of lymphocyte infiltration by the Chisholm standard.5 According to the European SS disease assessment index?ESSDAI?established by European League Against Rheumatism?EULAR?in 2009,we assessed the disease activity index of pSS patients.6 SPSS21.0 software was used for data analysis.Results:1 The expression of CD19⁺CD24^hiCD38^hiBregs and IL-17 in pSS patients:In pSS group,the CD19⁺CD24^hiCD38^hiBregs occupied 4.18 ±1.98%of CD19+cells,significantly higher than those in the control group which were1.10±0.63%?P<0.001?.The levels of serum IL-17 in pSS patients and health individuals were 17.99±9.88pg/ml and 13.32±5.29pg/ml respectively.There was statistic significance for the difference between the two groups?P< 0.05?;2 The expression of CD19⁺CD24^hiCD38^hiBregs and IL-17 in pSS patients with and without organ involvement: There were 20 cases with organ involvement,10 without organ involvement.The percentage of CD19+CD24hi CD38 hiBregs in the two groups were 4.29±2.27% and 3.96±1.33% and IL-17 were 19.04±9.93pg/ml and 15.50±9.47pg/ml.The expressions of both CD19⁺CD24^hiCD38^hiBregs and IL-17 were no differences in pSS patients with and without organ involvement?P>0.05?;3 The expression of CD19⁺CD24^hiCD38^hiBregs and IL-17 in pSS patients with and without autoantibodies: There were 2 cases with ANA negative,28 ANA positive;21 RF negative,9 RF positive;14 anti-SSA negative,16 anti-SSA positive;23 anti-SSB negative,7 anti-SSB positive.The percentages of CD19+CD24hi CD38hiBregs?4.86± 2.21% and6.08±2.08%?inanti-SSA and anti-SSB positive groups were higher than those?3.40±1.39%and 3.60±2.08%?in anti-SSA and anti-SSB negative groups?t=2.22,3.37respectively;both P<0.001?.For the levels of serum IL-17 there were no difference between the two groups?P>0.05?;4 The correlation of CD19⁺CD24^hiCD38^hiBregs and IL-17 with clinical indicators in pSS patients: The percentage of CD19⁺CD24^hiCD38^hiBregs was positively correlated with the level of IgG?r=0.36,P<0.05?and IgM?r=0.45,P<0.05?,while the percentage of CD19⁺CD24^hiCD38^hiBregs was negatively correlated with course of the disease?r=-0.37,P<0.05?and the level of hemoglobin?r=-0.43,P<0.05?in pSS.There were no correlation between the percentages of CD19+CD24hi CD38 hiBregs and other clinical indicators.The level of serum IL-17 was positively correlation with ESR?r=0.58,P<0.001?and WBC?r=0.53,P<0.01?;5 The expression of CD19⁺CD24^hiCD38^hiBregs and IL-17 in pSS patients with different grades of salivary gland tissues: According to the Chisholm standard,there were 6 cases with grade ? and 24 with grade?in 30 pSS patients.The percentages of CD19⁺CD24^hiCD38^hiBregs were 3.23±1.42%and 4.41±2.05% respectively in the two groups,while the levels of serum IL-17 were 17.97±8.35pg/ml and 17.83±10.24pg/ml.There were no significant differences of both expression of CD19⁺CD24^hiCD38^hiBregs and IL-17 in the pSS patients with different Chisholm grades?P>0.05?;6 The expression of CD19⁺CD24^hiCD38^hiBregs and IL-17 in pSS patients with different degrees of ESSDAI: According to ESSDAI,the pSS patients were divided into three groups,score 0-2 with 9 cases,score 3-5 18 and score 6-8 3 respectively.The percentages of CD19⁺CD24^hiCD38^hiBregs were 3.29±1.48%,4.34±2.02% and 5.84±1.40% respectively in the three groups,while the levels of serum IL-17 were 16.30±7.75pg/ml,17.51±9.76pg/ml and 24.62±15.80pg/ml.The differences in both percentages of CD19+CD24hiCD38hi Bregs and levels of serum IL-17 had no statistical significance among the three groups?all P>0.05?;7 The correlation between CD19⁺CD24^hiCD38^hiBregs and serum IL-17:statistical analysis showed that r=0.21,P=0.28.This indicated that there was no correlation between the level of serum IL-17 and the percentage of CD19⁺CD24^hiCD38^hiBregs in pSS patients?P>0.05?.Conclusion:1 CD19⁺CD24^hiCD38^hiBregs and IL-17 in pSS patients were both expressed abnormally,higher than those in healthy individuals.It suggested that CD19⁺CD24^hiCD38^hiBregs and IL-17 take part in the pathogenesis of pSS.2 The expression of CD19+CD24hiCD38hi Bregs was higher in pSS patients with anti-SSA antibody and anti-SSB antibody than those without anti-SSA and anti-SSB antibody,and the Bregs was positively correlated with IgG and IgM.Those indicated that the CD19⁺CD24^hiCD38^hiBregs may be involved in the immune disorders in pSS.3 The level of serum IL-17 was positively associated with ESR and WBC in pSS patients.It suggested that IL-17 could be an indicator for monitoring disease activity.