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【Objective】 To establish an effective method for acquiring bone marrow-derived dendritic cells (DCs) from BALB/C mice in vitro and to establish a reservoir of DC precursor cells. 【Methods】 CD117+ hematopoietic stem cells (HSCs) were isolated and purified from bone marrow of BALB/C mice by immunomagnetic beads separation system (MACS), and then amplified in vitro with mouse stem cell factor (SCF) and interleukin-3 (IL-3). HSC was induced to differentiate into DCs by adding granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and IL-4. Different cytokines (tumor necrosis factor-alpha or IL-10) were added to control the maturity of dendritic cells. Then the morphology (electron microscopy), surface molecular markers (FACS method) and cytokine secretion level (ELISA method) were identified. 【Results】 ① The purity of CD117 + HSC isolated and purified by MACS system was over 95%. ② SCF plus IL-3 could effectively stimulate HSC amplification. ③ The morphology of mature DC (mDC) and immature DC (imDC) was significantly different under light and scanning electron microscopy. ④ In the expressions of surface markers CD40, CD80, CD86, I-A/I-E, there were significant differences between imDC group and mDC group (P<0.01). ⑤ After LPS stimulation, the secretion of IL-12 in imDC group did not change significantly (P=0.064), while the secretion of IL-12 in mDC group increased significantly (P=0.009). LPS and TNF-α had a synergistic effect in stimulating DC maturation. 【Conclusion】 Specific combinations of cytokines can effectively induce the differentiation of bone marrow HSCs into DCs in BALB/C mice, and can control the maturity of DCs. This study makes it possible to use gene modified dendritic cells in GD immunotherapy.
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Objective To assess the disorders of glucose metabolism and insulin resistance in patients with rheumatoid arthritis (RA) and its relationship with disease activity.Methods One hundred and twenty-three RA patients along with 98 age and sex matched controls were studied.Seventy-five g oral glucose tolerance test was performed.The homeostasis model assessment of insulin resistance (HOMA-IR) and beta cell function (HOMA-β) were evaluated.Disease activity score (DAS28) was used to assess disease activity.According to their DAS28 values,patients were divided into high disease activity group and low to moderate disease activity group.Glucose tolerance and HOMA-IR were compared between the two groups.Parameters that reflects disease activity,such as CRP and ESR,as well as disease activity scores were compared between patients with T2DM or prediabetes and patients with normal glucose tolerance.The data was analyzed by t test,Pearson correlation analysis and chi-square test.Results The prevalence of T2DM [20.3％(25/123) vs 5.1％ (5/98),x2=10.774,P＜0.01] and prediabetes [39.0％ (48/123) vs 7.1％ (7/98),x2=29.657,P＜0.01] increased in RA patients compared to controls.RA patients had higher HOMA-IR (2.5±1.5 vs 0.8±0.4; t=5.185,P＜0.01) and lower HOMA-β (83±69 vs 192±85; t=3.768,P＜0.01) compared to controls.ESR [(55±30) mm/1 h vs (37±26) mm/1 h; t=3.159,P＜0.01],CRP [(40±23) mg/L vs (19±10) mg/L; t=3.628,P＜0.01] and DAS28 score (5.6±1.3 vs 4.8±1.2; t=2.923,P＜0.01) were higher in RA patients with T2DM or prediabetes than in RA patients with normal glucose tolerance.In RA patients,the HOMA-IR was significantly positively correlated with DAS28 (r=0.39,P＜0.01),ESR (r=0.54,P＜0.01)and CRP (r=0.20,P＜0.05).The HOMA-IR value and fasting insulin levels were higher in high disease activity patients (DAS28＞ 5.5) than in low-to-moderate disease activity patients (DAS28 ≤5.5) although fasting plasma glucose level did not differ significantly in these two groups.Conclusion The prevalence of T2DM and prediabetes increases in RA patients comparing to controls.RA patients have insulin resistance that is associated with disease activity and systemic inflammation.