Expression And Significance Of CD19⁺CD5⁺CD1d^hi Regulatory B Cell And APRIL In Peripheral Blood Of Patients With Primary Sj???gren''s Syndrome

Objective : primary sj?gren's syndrome is an autoimmune connective tissue disease with external exocrine glands dysfunction especially lacrimal glands and salivary glands.The essential characteristic is the lymphocytes infiltration highly within the tissues and organs involved.Clinically,the most frequent symptom is dryness of eyes and mouth,and the patients always have multiorgan involvement including lung,liver,kidneys,musculoskeletal symptoms,nervous and blood systemic manifestations.The etiology and pathogenesis of pSS are not clear.Studies have found that the onset of pSS is associated with genetic factors,environment,different cells and cytokines.In recent years,research on Bregs in pSS is a hot spot.This study was performed to investigate the expression of CD19⁺CD5⁺CD1d^hiBreg and APRIL in pSS patients,analysis the correlation of the expression levels with immunoglobulins,autoantibodies,labial gland pathological grades,disease activity;and to provide a new diagnostic method to diagnose and a new treatment target for pSS.Methods:1 The 30 pSS patients were selected from April 2015 to October 2015 in Department of Rheumatology and Clinical Immunology of Hebei General Hospital,as experimental group.All the patients were fulfilled with 2002 European classification criteria for sj?gren's syndrome,without other connective tissue diseases,HCV,AIDS,lymphoma and so on.20 cases of normal healthy individuals were enrolled as control group.2 The expression of CD19⁺CD5⁺CD1d^hiBregs from peripheral blood mononuclear cells?PBMCs?was detected by flow cytometry?FCM?.ELISA was used to determine the levels of serum APRIL in both groups.We selectedthe basic information and clinical data of experimental subjects including WBC,RBC,Plt,Hb,ANA,ENAs,RF,GLB,Igs,CRP,ESR,C3,C4.3 Labial salivary gland tissues were stained by HE and graded by the degree of lymphocyte infiltration according to the Chisholm standard.4 According to the SS Disease assessment index?ESSDAI?established by European League Against Rheumatism?EULAR?in 2009,we assessed the disease activity index of pSS patients.5 SPSS17.0 software was used for data analysis.Results:1 Expression of CD19⁺CD5⁺CD1d^hi Bregs and APRIL in pSS patients:In experimental group,the percentage of CD19⁺CD5⁺CD1d^hiBregs occupied 5.06±0.69% of CD19⁺cells,significantly higher than that in the control group which was 2.55±0.54%?P<0.001?.The levels of serum APRIL in pSS patients and health individuals were 7.40±2.20 ng/ml and 1.99±0.44 ng/ml respectively.There was statistics significance between differences of the two groups?P<0.001?.2 Expression of CDl9⁺CD5⁺CD1d^hi Bregs and APRIL in pSS patients with and without autoantibodies: There were 7 cases with ANA negative,23 with ANA positive;19 with RF negative,11 with RF positive;11 with anti-SSA negative,19 with anti-SSA positive;20 with anti-SSB negative,10 with anti-SSB positive.The percentages of CD19⁺CD5⁺CD1d^hiBreg?5.44±0.46%,5.59±0.53%?in anti-SSA and anti-SSB positive groups were higher than that?4.41±0.49%,4.79±0.61%?in anti-SSA,anti-SSB antibody negative groups?t=-5.70,-3.53;P<0.001?.The levels of serum APRIL were no difference between the two groups?P>0.05?.3 Correlation among CD19⁺CD5⁺CD1d^hiBregs,APRIL and clinical indicators in pSS patients: the percentage of CD19⁺CD5⁺CD1d^hi Breg was positively correlated with IgG?r=0.44;P<0.05?,the percentage of CD19⁺CD5⁺CD1d^hiBregs was negatively correlated with age?r=-0.43;P<0.05?in pSS.There were no correlation between the percentages of CD19⁺CD5⁺CD1d^hiBregs and other clinical indicators.The level of serum APRIL waspositively correlation with ESR?r=0.91;P<0.05?.4 Expression of CDl9⁺CD5⁺CD1d^hi Bregs and APRIL in pSS patients with different grades of salivary gland tissues: there were 8 cases with ?grade of salivary gland tissues in pSS,22 with ?grade.The percentages of CD19⁺CD5⁺CD1d^hiBregs were 4.84±0.69%,5.14±0.68% respectively and levels of serum APRIL were 6.90±1.84ng/ml,7.59±2.34ng/ml respectively.There were no significant differences between the groups?t=-1.07,-0.75;P>0.05?.5 Expression of CDl9⁺CD5⁺CD1d^hi Bregs and APRIL in pSS patients with different degrees of ESSDAI: according to ESSDAI,we divided pSS patients into three groups with 6,19 and 5 cases respectively.The percentages of CDl9⁺CD5⁺ CD1 dhiBregs were 4.95±0.78%,5.09±0.68%,5.08±0.76%.The levels of serum APRIL were 8.73±3.11ng/ml,6.76±1.69 ng/ml,8.27±2.17ng/ml.The levels had no differences among the three groups?F=0.10,0.93;P>0.05?.6 Correlation between CD19⁺CD5⁺CD1d^hiBregs and serum APRIL:There was no correlation between the level of serum APRIL and the percentage of CD19⁺CD5⁺CD1d^hiBregs?r=-0.04;P>0.05?.Conclusion:1 CD19⁺CD5⁺CD1d^hiBregs and APRIL in pSS patients were both expressed abnormally,higher than healthy individuals.It suggested that the onset of pSS was associated with abnormal B cells sub-group CD19⁺CD5⁺CD1d^hiBregs and serum APRIL.2 The expression of CD19⁺CD5⁺CD1d^hiBregs was higher in pSS patients with anti-SSA antibody than those without anti-SSA;compared with anti-SSB antibody negative pSS patients,the expression of CD19⁺CD5⁺CD1d^hiBregs was higher.The percentage of CD19⁺CD5⁺CD1d^hiBregs was positively correlated with IgG.It indicated that the increase of CD19⁺CD5⁺CD1d^hiBregs maybe associated with autoantibody production.3 The level of serum APRIL was positively associated with ESR.It could be served as an inflammatory marker for monitoring the activities ofpSS.