树突状细胞、炎性单核细胞及巨噬细胞在小鼠感染肺孢子菌过程中变化的研究  

Alterations of dendritic cells, inflammatory monocytes and macrophages in mice during Pneumocystis ;murina infection

作　　者：杨爽利[1] 胡旸[1] 王栋[1] 郭斐[1] 吴秀芝[1] 刘雅兰[1] 翟侃[1] 童朝晖[1] 

机构地区：[1]首都医科大学附属北京朝阳医院呼吸与危重症医学科,100020

出　　处：《中华微生物学和免疫学杂志》2015年第5期328-334,共7页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金（81370102）;北京市教育委员会科技计划重点项目（KZ20140025026）

摘　　要：目的：研究非免疫抑制小鼠感染肺孢子菌（ Pneumocystic murina，P．murina）过程中树突状细胞、炎性单核细胞以及巨噬细胞的变化和表型特征，以探讨上述细胞在肺孢子菌感染时发挥的作用。方法选择6～8周的雄性野生型C57BL／6小鼠构建肺孢子菌感染动物模型，分为实验组和手术对照组，并设空白对照组。肺孢子菌感染后1周、2周、3周和4周，采用TaqMan实时荧光定量PCR法检测肺孢子菌载量；制作病理切片观察肺组织的炎症程度；采用流式细胞术检测两组小鼠肺脏、外周血和骨髓中树突状细胞、炎性单核细胞和巨噬细胞以及脾脏中炎性单核细胞的数量变化，并检测主要组织相容复合物（ major histocompatability complex， MHC）Ⅱ和CX3C趋化因子受体（ CX3C chemokine receptor， CX3CR1）-1，低表达CC趋化因子受体（ CC chemokine receptor， CCR）-2在感染后肺内各细胞群的表达情况。结果实验组小鼠菌载量呈现先升高再降低的趋势，但差异无统计学意义（ P＞0．05）。肺孢子菌感染3周和4周时，肺组织病理改变较重，可见到肺泡结构不清，间质内有炎性细胞浸润以致肺泡间隔增厚。肺孢子菌感染后，肺内CD11 c＋CD11 b＋树突状细胞数量增高，外周血中CD11c＋CD11b＋树突状细胞数量降低，外周血中的炎性单核细胞数量先降低后增高，差异均有统计学意义（P＜0．05）；但肺内的巨噬细胞的数量变化无统计学意义（P＞0．05）。 CD11c＋CD11b＋树突状细胞高表达MHCⅡ、CX3CR1，少量表达CCR2；炎性单核细胞高表达CCR2，部分表达MHCⅡ，低表达CX3CR1；巨噬细胞高表达CX3CR1，低表达MHCⅡ和CCR2。结论非免疫抑制小鼠感染肺孢子菌后，肺内CD11c＋CD11b＋树突状细胞募集增多，高表达MHCⅡ，参与了肺孢子菌感染的免疫应答。Objective To investigate the alterations and phenotypes of dendritic cells, inflamma-tory monocytes and macrophages in immunocompetent mice during Pneumocystis murina （ P.murina） infec-tion for further analysis of the function of these cells during P.murina infection.Methods Wild type male C57BL/6 mice at age 6-8 weeks were randomly divided into two groups including the group with P.murina infection and the group receiving sham surgery.The mice without any intervention were used to set up the blank control group.The loads of P.murina strains in lung tissues of each mouse were quantified by TaqMan real-time fluorescence polymerase chain reaction after the infection.Histopathological examination was per-formed to evaluate the degree of inflammation in lung tissues.The numbers of dendritic cells, inflammatory monocytes and macrophages in lung tissues, peripheral blood and bone marrow samples, and the changes of inflammatory monocytes in spleen tissues were measured by flow cytometry analysis.The expression of major histocompatability complexⅡ（MHCⅡ）, CX3C chemokine receptor 1 （CX3CR1） and CC chemokine re-ceptor 2 （ CCR2 ） by dendritic cells, inflammatory monocytes and macrophages in lung tissues during P.murina infection were analyzed by flow cytometry analysis.All of the data were collected one, two, three and four weeks after the corresponding treatments.Results The loads of P.murina strains in P.murina in-fected mice were elevated after two and three weeks infection, but decline at week 4 （P〉0.05）.Significant pathological changes including the alveolar destruction, inflammatory cell infiltration and thickened alveolar septum in mice with P.murina infection were observed under a microscope at week 3 and week 4.Comparednbsp;to the sham surgery treatment group, the number of CD11c＋CD11b＋dendritic cells were increased in lung tissues, but decreased in blood samples during P.murina infection （ P〈0.05） .The levels of inflammatory monocytes in blood samples fell at week 3 and then rose

关 键 词：肺孢子菌感染 树突状细胞 炎性单核细胞 巨噬细胞 

分 类 号：R519[医药卫生—内科学]