Objective To analyze the effect of cyclosporin A(CsA), rapamycin(RPM) and macophenolic acid(MPA) on the co-stimulated lymphocytes, CD28 and CD40, and their production of Th1/Th2 cytokine, IL-2, IFN-γ, IL-4, IL-10 and IL-12. Methods The experimental groups were divided into ①mono-stimulating and co-stimulating groups: CD3 mAb mono-stimulating (group a),CD3 mAb+CD28 mAb co-stimulating (group b), CD3 mAb+CD28 mAb+CD40 L mAb co-stimulating(group c), CD3 mAb+CD28 mAb+CTLA4 mAb co-stimulating (group d). ②CsA groups: 300 ng/ml of CsA was added to group a, b, c and d. ③RPM groups: 300 ng/ml of RPM was added to group a,b, c and d. ④MPA groups:300 ng/ml of MPA was added to group a, b, c and d. The cytokine production was measured by ELISA.Results The co-stimulated CD28 and CD40 Th1/Th2 cytokines production of IFN-γ, IL-2, IL-4 and IL-10 were significantly increased. Compared with group a, IFN-γ increased from (248.91±11.20)ng/ml to (555.08±24.42)ng/ml and (548.19±33.06)ng/ml, IL-2 increased from (29.48±8.61)ng/ml to (1100.82±99.29)ng/ml and (842.23±29.31)ng/ml, IL-4 increased from (32.29±6.76)ng/ml to (116.02±15.03)ng/ml and (147.28±18.07)ng/ml, IL-10 increased from (147.01±10.47)ng/ml to (291.79±12.47)ng/ml and (302.52±35.18)ng/ml,respectively, P＜0. 01. Compared group b with group c, the Th1 cytokines production was decreased.IL-2 and IL-12 decreased (P＜0.05). The Th2 cytokine IL-4 production was increased (P＜0. 05).CTLA4 mAb and three other immunosuppressants, CsA, RPM and MPA, inhibited co-stimulated lymphocyte's both cytokines Th1 and Th2 production. The inhibitory effect of CsA on Th1/Th2 cytokine production was more significant than RPM and MPA did. The co-inhibitory effect of CTLA4 mAb and CsA was observed as well. The increased co-stimulated CD28 and CD40 IL-12 production could be suppressed by MPA. CsA and RPM had no inhibitory effect on the IL-12 production.Conclusions CD28/CD40 co-stimulatory pathway plays the key role in lymphocyte activation and Th1/Th2 cytokine production. CsA, RPM and MPA can inhibit co-stimulated lymphocyte's Th1 and Th2 cytokine production. CsA and CTLA4 mAb have co-inhibitory effect on co-stimulated lymphocyte's Th1/Th2 cytokines production. CD40 L mAb decreases the Th1 cytokines production(including IL-12) and increases the Th2 (mainly IL-4) production, which may be the mechanism of its longevity effect on allograft.

Objective To assess the expression and significance of proprotein convertase subtilisin kexin 9 (PCSK9) in patients with rheumatoid arthritis (RA).Methods Sixty-five RA patients and forty-seven healthy controls were recruited in this study.The body mass index (BMI) and serum total cholesterol (TC),triglyceride (TG),high density lipoprotein (HDL),lipoprotein a,low density lipoprotein (LDL),very low density lipoprotein(VLDL),apolipoprotein A (ApoA),apolipoprotein B (ApoB) and the ratio of LDL-C/HDL-C were tested.Other parameters included disease activity score 28 (DAS28),rheumatoid factor (RF),anti-cyclic citrullinated peptide (CCP) antibody,erythrocyte sedimentation rate (ESR),c-reactive protein (CRP).Serum PCSK9 level was measured by ELISA and compared between RA patients and healthy controls.Results (1) The serum PCSK9 levels in RA patients were higher than those in healthy controls [(409.36 ±223.52) μg/L vs (292.19 ± 109.79) μg/L,P ＜ 0.05].(2) Compared with subgroup of moderate and low active disease and patients in remission,PCSK9 was significantly higher in patients with highly active disease (P ＜ 0.05).(3) The serum PCSK9 levels were positively correlated with RF,TC,TG,LDL,very low density lipoprotein (VLDL),ApoB,with r values as 0.303,0.490,0.320,0.451,0.319,0.463,respectively (P ＜ 0.05).(4) Multiple stepwise regression analysis showed that DAS28,RF,TC and LDL-C/HDL-C were relevant factors for PCSK9 in RA patients.Conclusions The serum PCSK9 level is elevated in RA patients,which is related to RF,disease activity,TC,TG,LDL,VLDL,ApoB.This suggests that PCSK9 is potentially linked to inflammatory reaction and lipid metabolism in rheumatoid arthritis.