The Study Of The Role Of NK-22 Cells And Related Functional Molecules In The Pathogenesis Of Autoimmune Diseases

Objective: To investigate the role of natural killer cell-22 and interleukin-22 in the pathogenesis of rheumatoid arthritis(RA)and systemic lupus erythematosus(SLE),and to analyze their correlation with clinical indicators.Methods: Using CD3-,CD56+,NKp44+ and CCR6+ as markers,the percentages of peripheral blood NK-22 cells were detected by flow cytometry four-color fluorescence in 49 patients with active RA,9 patients with stable RA,24 healthy controls(HC),19 patients with active SLE,19 patients with stable SLE and 17 healthy controls(HC).The levels of IL-22 and TRAP in peripheral blood supernatant of 58 patients with RA(10 males and 48 females),38 patients with SLE(6 males and 32 females)and 41 healthy controls(10 males and 31 females)were measured by ELISA.Gender,age and course of disease were recorded,and rheumatoid factor(RF),anti-CCP antibody,hypersensitive C-reactive protein(hs CRP),erythrocyte sedimentation rate(ESR),lymphocyte subsets(CD3+cells,CD3+CD4+cells,CD3+CD8+cells,C D19+cells,CD16+CD56+cells),total type I collagen amino-terminal prolonging peptide(PIPN),C-terminal cross-linking telopeptide of type I collagen(?-CTX)were detected in RA patients.All the patients took X-ray examinations.The disease activity score in 28 joints(DAS28)was calculated.The correlation of NK-22 cells and IL-22 with clinical indicators and TRAP concentrations was analyzed,and the risk factors of bone destruction in RA patients were analyzed.Gender,age and course of disease were recorded,and blood routine,hs CRP,ESR,circulating immune complex(CIC),immunoglobulin(Ig G,Ig A,Ig M),lymphocyte subsets,bone turnover indexes(PIPN,?-CTX)were detected in SLE patients.And the systemic lupus erythematosus disease activity index(SLEDAI)was calculated.The correlation of NK-22 cells and IL-22 with clinical indicators was analyzed in SLE patients.T-test was used for comparison between the two groups.Multigroup measurements were compared by ANOVA.The correlation analysis was performed by Pearson linear correlation analysis.The risk factors of bone destruction were analyzed by unconditional multivariate logistic regression.Results: 1.The proportions of NK-22 cells in peripheral blood of active RA patients,stable RA patients and healthy controls were(0.274�0.084)%,(0.144 �0.039)% and(0.004�0.020)% respectively.The proportion of NK-22 cells in peripheral blood of active RA patients was higher than that in stable and healthy controls(P<0.01).The concentrations of IL-22 in peripheral blood of active RA patients,stable RA patients and healthy controls were(17.36�3.72)ng/L,(12.44�0.36)ng/L and(10.35�3.61)ng/L respectively.The concentration of IL-22 in peripheral blood of active RA patients was higher than that in stable and healthy controls(P<0.01).The concentrations of TRAP in peripheral blood of active RA patients,stable RA patients and healthy controls were(9.04�3.69)ng/L,(4.25�0.91)ng/L and(3.24�1.29)ng/L respectively.The concentration of TRAP in peripheral blood of active RA patients was higher than that in stable and healthy controls(P<0.01).The proportion of NK-22 cells in peripheral blood of RA patients was positively correlated with ESR,hs CRP,RF,DAS28 score,the proportion of CD3+C D4+cells,IL-22 and TRAP expression levels(the r values were respectively 0.890,0.306,0.850,0.831,0.388,0.961,0.295,P<0.05).The expression level of IL-22 in peripheral blood of RA patients was positively correlated with ESR,hs CRP,RF,DAS28 score and the proportion of CD3+CD4+cells((the r values were respectively 0.883,0.292,0.875,0.841,0.369,P<0.05).The expression level of TRAP in peripheral blood of RA patients was positively correlated with DAS28 score(r=0.329,P<0.05).The percentage of NK-22 cells in anti-CCP antibody positive group(n=33)was higher than that in anti-CCP antibody negative group(n=16),which was respectively(0.31�0.08)% and(0.20�0.04)%.There was significant d ifference between the two groups(P<0.05).The concentration of IL-22 in anti-CCP antibody positive group(n=33)was higher than that in anti-CCP antibody negative group(n=16),which was respectively(18.74�3.75)ng/L and(14.50�1.20)ng/L.There was significant difference between the two groups(P<0.05).The concentration of TRAP in anti-CCP antibody positive group and anti-CCP antibody negative group were respectively(9.59�3.73)ng/L and(7.91�3.44)ng/L.There was no significant difference between the two groups(P>0.05).Multivariate logistic regression analysis showed that TRAP and IL-22 in RA patients had an influence on the occurrence of imaging bone destruction(P<0.05),and OR values were respectively 1.31(1.05,1.63)and 0.49(0.25,0.97).NK-22 cells proportion,IL-22 concentration and TRAP concentration in peripheral blood of RA patients were in no correlation with age,course of disease,PIPN,?-CTX,CD3+cells,CD3+CD8+cells,CD19+cells and CD16+CD56+ cells.2.The proportions of NK-22 cells in peripheral blood of active SLE patients,stable SLE patients and healthy controls were(0.45�0.11)%,(0.24�0.04)% and(0.01�0.02)% respectively.The proportion of NK-22 cells in peripheral blood of active SLE patients was higher than that in stable and healthy controls(P<0.01).The concentrations of IL-22 in peripheral blood of active SLE patients,stable SLE patients and healthy controls were(4.08�1.51)ng/L,(6.51�3.63)ng/L and(9.13�4.78)ng/L respectively.The concentration of IL-22 in peripheral blood of active SLE patients was lower than that in stable and healthy controls(P<0.01).The concentrations of TRAP in peripheral blood of active SLE patients,stable SLE patients and healthy controls were(4.92�2.75)ng/L,(4.30�1.74)ng/L and(2.45�0.61)ng/L respectively.The concentration of TRAP in peripheral blood of active SLE patients was higher than that in stable and healthy controls(P<0.01).The proportion of NK-22 cells in peripheral blood of SLE patients ware positively correlated with CIC and Ig G(the r values were respectively 0.464 and 0.457,P<0.05).The expression level of IL-22 in peripheral blood of SLE patients was negatively correlated with SLEDAI score(r=-0.719,P=0.001).NK-22 cells proportion,IL-22 concentration and TRAP concentration in peripheral blood of SLE patients was in no correlation with gender,age,course of disease,ESR,hs C RP,Ig A,Ig M,CD3+cells,CD3+C D4+cells,CD3+CD8+cells,CD19+cells,CD16+CD56+ cells,PIPN and ?-CTX.Conclusion: 1.The percentage of NK-22 cells and the concentrations of IL-22 and TRAP are increased,in peripheral blood of patients with active RA.All of them are positively correlated with DAS28 score,suggesting that NK-22 cells,IL-22 and TRAP are correlated with the activity of RA.NK-22 cells and IL-22 are positively correlated with RF,and the proportion of NK-22 cells and the concentration of IL-22 in anti-CCP antibody positive group are higher than that in anti-CCP antibody negative group,suggesting that NK-22 cells and IL-22 may be related to the severity and prognosis of RA.IL-22 and TRAP are risk factors for bone destruction.2.The proportion of NK-22 cells in peripheral blood of patients with active SLE are increased,and are positively correlated with CIC and Ig G,suggesting that NK-22 cells may be related to the immune imbalance of SLE.The concentration of IL-22 in peripheral blood of SLE patients are decreased,and the concentration of IL-22 are negatively correlated with SLEDAI score,suggesting that IL-22 may play a protective role in the pathogenesis of SLE.