尘螨过敏性支气管哮喘患者肺泡巨噬细胞表面CD86的表达及抗CD86单抗对其炎性细胞因子生成的作用

目的　观察尘螨过敏性支气管哮喘 (简称哮喘 )患者肺泡巨噬细胞表面CD86的表达 ,探讨抗CD86单抗抑制哮喘炎症反应的机制。方法　从 10例尘螨过敏性哮喘患者支气管肺泡灌洗液(BALF)和外周血中分离出肺泡巨噬细胞 (AM)和CD 4 T淋巴细胞 ,AM经螨变应原的刺激活化后与CD 4 T淋巴细胞共培养。每例患者的培养细胞分为抗CD86单抗干预组和CD86表达组 ,每组内再分为实验组和对照组。抗CD86单抗干预组、实验组分别加抗CD86单抗至 0 1mg/L(低剂量 )和 1.0mg/L(高剂量 )两个剂量 ,对照组不加相应抗体。共培养 72h后分别留取培养上清液 ,用酶联免疫吸附测定(ELISA)法检测细胞因子γ干扰素 (INF γ)、白细胞介素 4 (IL 4 )、IL 5的产生量。CD86表达组、实验组加螨变应原 ,对照组不加变应原 ,共培养 2 4h后收集培养细胞 ,流式细胞仪 (FCM)检测AM表面CD86分子的表达水平。结果　AM在尘螨变应原的刺激活化下 ,实验组AM表面CD86分子平均荧光密度表达水平为 (5 0± 9) % ,对照组为 (2 3± 5 ) % ,两组比较差异有显著性 (P <0 0 1) ;实验低剂量组IL 4、IL 5的分泌量分别为 (135± 19)ng/L、(10 4± 2 1)ng/L ,实验高剂量组分别为 (90± 17)ng/L、(6 8± 14 )ng/L ,对照组IL 4、IL 5的分泌量分别为 (187± 2 4 )ng/L、(16

CD86 expression on alveolar macrophages and effects of anti-CD86 monoclonal antibody on the production of inflammatory cytokines in dust-mite allergic asthma patients

OBJECTIVE: To investigate the CD(86) molecule expression on alveolar macrophages (AMs) before and after AM activation with dust-mite allergen and effects of anti-CD(86) monoclonal antibody on the production of inflammatory cytokines produced by cultured CD(34)(+) T cells, and to explore the pathogenesis of anti-CD(86) monoclonal antibody on inhibiting asthmatic inflammation. METHODS: AMs and CD(34)(+) T cells were isolated from bronchoalveolar lavage fluid (BALF) and from peripheral blood derived from 10 asthma patients allergic to house dust-mite dermatophagoides pteronyssinus. AMs were stimulated and activated by house dust-mite allergen and co-cultured with CD(34)(+) T cells. The co-cultured cells were divided into anti-CD(86) monoclonal antibody intervention group and CD(86) expression group with which the control groups were set up. The anti-CD(86) antibody was added to 0.1 mg/L and 1.0 mg/L in the experiment group and no antibody was added in the control group. The supernatants were harvested after 72 hour co-culture. The concentration of interferon-gamma (INF-gamma), interleukin-4 (IL-4) and IL-5 were determined by enzyme linked immunosorbent assay (ELISA). CD(86) expression group was divided into experiment and control groups, and house dust-mite allergen was added to the experiment group but no allergen was added in the control group. The cells of the two groups were harvested after 24 hour co-culture. The CD(86) molecule expression on the AMs was detected by flow cytometry. RESULTS: After activated by house dust-mite allergen, the average fluorescence density of CD(86) on AM in the experiment group [(50 +/- 9)%] was significantly higher than that in the control group [(23 +/- 5)%, P < 0.01]. The production of IL-4 and IL-5 was (135 +/- 19) ng/L, and (104 +/- 21) ng/L in the low-dose group respectively; (90 +/- 17) ng/L and (68 +/- 14) ng/L in the high-dose group; (187 +/- 24) ng/L and (161 +/- 23) ng/L in the control group; the differences among groups were significant (P < 0.01). The production of IFN-gamma was (193 +/- 39) ng/L in the low-dose group and (201 +/- 47) ng/L in the high-dose group respectively, and there was no significant difference between the two groups (P > 0.05). CONCLUSION: AMs may effectively present allergen to T cells in allergic asthma patients. Anti-CD(86) monoclonal antibody can inhibit the production of IL-4 and IL-5 by blocking the co-stimulatory signal of CD(34)(+) T cells. These data suggest that anti-CD(86) monoclonal antibody may hold therapeutic potentials in asthma by inhibiting airway inflammation.