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目的探讨负载卵清蛋白(OVA)的树突细胞回输至小鼠体内对过敏性哮喘的影响。方法将30只雄性BALB/c小鼠采用随机数字表法分为3组,每组10只。健康对照组不做任何处理;哮喘组给予磷酸盐缓冲液(PBS),然后致敏和激发;OVA-DC干预组尾静脉注射负载OVA的树突细胞1×106个,然后致敏和激发。将小鼠骨髓细胞用重组小鼠粒系-巨核系集落刺激因子(rmGM-CSF)和重组白介素4(rmIL-4)诱导分化为树突细胞,并通过形态学和流式细胞术检测细胞表面的CD11c予以鉴定。用OVA和脂多糖(LPS)刺激后,流式细胞术检测细胞表面的主要组织相容性复合体Ⅱ类分子(MHCⅡ)和CD86,以此判断树突细胞的成熟状态;混合淋巴细胞增殖试验(MLR)检测不同状态的树突细胞刺激淋巴细胞增殖的能力。通过尾静脉注射树突细胞至小鼠体内,然后在特定时间进行致敏和激发,观察小鼠的生存状态、测定脾脏质量;激发后24 h收集小鼠肺泡灌洗液检测组胺水平;取肺组织行苏木素-伊红(HE)染色,镜检观察肺部炎症严重程度。结果小鼠骨髓细胞经rmGM-CSF和IL-4诱导分化为树突细胞,树突细胞在负载OVA后免疫表型未发生明显的变化,仍处于未成熟状态,刺激淋巴细胞增殖的能力也很微弱。3组脾脏质量和肺泡灌洗液组胺水平比较,差异有统计学意义(P<0.05),OVA-DC干预组较健康对照组和哮喘组升高,哮喘组较健康对照组升高(P<0.05)。HE染色显示OVA-DC干预组小鼠肺组织气道管壁增厚,周围有大量炎性细胞浸润;哮喘组小鼠炎性反应较轻;健康对照组小鼠未发现炎性反应的特征。结论负载OVA后的树突细胞通过尾静脉回输至小鼠体内可以促进过敏性哮喘的发生,树突细胞在过敏性哮喘的发生机制方面发挥重要的作用。
Objective To investigate the effect of ovalbumin (OVA) loaded dendritic cells on allergic asthma in mice. Methods Thirty male BALB / c mice were randomly divided into three groups (n = 10). The rats in the asthma group were given phosphate buffered saline (PBS) and then sensitized and challenged. OVA-DC group was injected with 1 × 106 dendritic cells loaded with OVA through tail vein, and then sensitized and challenged. The mouse bone marrow cells were induced to differentiate into dendritic cells using recombinant murine myeloid-megakaryocyte colony stimulating factor (rmGM-CSF) and recombinant interleukin-4 (rmIL-4), and the cell surface was detected by morphological and flow cytometry Of the CD11c be identified. After stimulation with OVA and lipopolysaccharide (LPS), flow cytometry was used to detect the major histocompatibility complex class II molecules (MHC II) and CD86 on the cell surface to determine the maturation status of dendritic cells; mixed lymphocyte proliferation assay (MLR) to detect the ability of different states of dendritic cells to stimulate lymphocyte proliferation. The dendritic cells were injected into the mice via the tail vein, then sensitized and challenged at a specific time, the survival state of the mice was observed, and the quality of the spleen was measured; the mouse alveolar lavage fluid was collected 24h to detect histamine levels; Tissue samples were stained with hematoxylin-eosin (HE), and the severity of lung inflammation was observed microscopically. Results The mouse bone marrow cells were induced to differentiate into dendritic cells by rmGM-CSF and IL-4. The immunophenotype of dendritic cells did not change significantly after loaded OVA, and was still immature, and the ability of stimulating lymphocyte proliferation was also very high weak. Compared with healthy control group and asthmatic group, the levels of spleen mass and histamine in bronchoalveolar lavage fluid of 3 groups were significantly different (P <0.05) <0.05). HE staining showed that the airway wall thickening in the lung tissue of OVA-DC-treated mice was accompanied by a large number of inflammatory cell infiltration. The inflammatory response in asthmatic mice was mild. No inflammatory reaction was found in healthy control mice. Conclusions Dendritic cells loaded with OVA are able to promote the development of allergic asthma through the tail vein transfusion into mice. Dendritic cells play an important role in the pathogenesis of allergic asthma.