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目的研究抗CD40配体单克隆抗体(CD40LmcAb)干预重症肌无力(MG)患者外周血单个核细胞(PBMC)的效果并探讨其机制。方法MG患者25例和健康对照组16名,分离培养外周血单个核细胞,分别用植物血凝素(PHA)和美洲商陆原(PWM)进行刺激,并用CD40LmcAb进行干预。PHA刺激组培养后收集上清检测γ干扰素(IFN γ)和白细胞介素4 (IL 4);PWM刺激组培养后收集上清检测抗乙酰胆碱受体抗体(AchRab)和抗突触前膜受体抗体(PsmRab)。比较MG患者两组中CD40LmcAb干预与否的差异并与健康对照组比较。结果MG患者PBMC体外诱生AchRab(0 32±0 11)、PsmRab(0 28±0 11)、IFN γ[ (36 24±10 47)pg/ml]和IL 4水平[ (263 08±35 95)pg/ml]对比健康对照组显著增高[分别为(0 10±0 02), (0 15±0 03), (17 56±2 94)pg/ml和(190 00±6 50)pg/ml,P<0 01),CD40LmcAb干预后MG组的上述四项指标均基本降至健康对照组水平(P>0 05)。结论CD40LmcAb在体外能有效干预MG患者PBMC诱生AchRab、PsmRab、IFN γ和IL 4的水平。
Objective To investigate the effect of anti-CD40 ligand monoclonal antibody (CD40LmcAb) on peripheral blood mononuclear cells (PBMC) in patients with myasthenia gravis (MG) and to explore its mechanism. Methods Twenty-five MG patients and 16 healthy controls were cultured and isolated. Peripheral blood mononuclear cells (PBMCs) were isolated and cultured. Phosphatidylinositol (PHA) and pokeweed (PPC) were respectively used for the stimulation and CD40LmcAb intervention. After stimulated with PHA, the supernatants were collected to detect IFN-γ and IL-4. After stimulated with PWM, the supernatants were collected and tested for anti-acetylcholine receptor antibody (AchRab) and anti-presynaptic membrane Antibody (PsmRab). Differences in the intervention of CD40LmcAb between the two MG groups were compared and compared with the healthy control group. RESULTS: PBMCs from MG patients were induced with AchRab (0 32 ± 0 11), PsmRab (0 28 ± 0 11), IFN γ [(36 24 ± 10 47) pg / ml] and IL 4 levels [263 08 ± 35 95 ) pg / ml] were significantly higher than those in healthy controls [(0 10 ± 0 02), (0 15 ± 0 03), (17 56 ± 2 94) pg / ml and (190 00 ± 6 50) pg / ml, P <0.01). After the intervention of CD40LmcAb, the above four indexes in MG group were all reduced to the level of healthy control group (P> 0.05). Conclusion CD40LmcAb can effectively induce the levels of AchRab, PsmRab, IFNγ and IL 4 induced by PBMCs in MG patients in vitro.