卡介苗接种对新生BALB／c小鼠脾树突状细胞表型和功能的影响  

Effect of neonatal BCG vaccination on phenotype and function of splenic dendritic cells of BALB/c mice

作　　者：刘春花[1] 刘恩梅[1] 刘崇海[2] 刘玮[2] 杨锡强[2] 李欣[3] 

机构地区：[1]重庆医科大学附属儿童医院呼吸科,400014 [2]重庆医科大学附属儿童医院免疫室,400014 [3]重庆医科大学附属儿童医院中心实验室,400014

出　　处：《中华儿科杂志》2008年第10期784-788,共5页Chinese Journal of Pediatrics

基　　金：国家自然科学基金委面上项目（30470760）,国家自然科学基金青年基金（30300321）,重庆市科委科技计划项目（2003-43-97）

摘　　要：目的探讨卡介苗（BCG）接种对新生BALB／c小鼠脾树突状细胞（DC）的作用，以明确BCG影响小鼠T细胞亚群分化发育的机制。方法BCG分别经腹腔、皮下接种新生鼠，经腹腔接种成年BALB／c小鼠。接种后4周：流式细胞仪检测脾DC表型；脾细胞体外用LPS或BCG再刺激培养后ELISA检测上清液细胞因子水平。结果（1）新生BALB／c小鼠腹腔接种BCG后，脾CD11C^＋CD8α^-DC百分比较对照组低[（45．00±14．14）％vs（67．00±8．27）％，P〈0．01]，相应的CD11C^＋CD8α^＋DC则较高；成年BALB／c小鼠腹腔接种BCG后，脾CD11C^＋CD8α^-DC百分比较对照组高[（70．00±5．21）％vs（57．00±6．22）％，P〈0．01]，CD11C^＋CD8α^＋DC则较低；同为腹腔接种BCG，成年BALB／c小鼠脾CD11C^＋CD8α^-DC百分比高于新生鼠，CD11C^＋CD8α^＋DC则低于新生鼠（P〈0．01）。（2）新生BALB／c小鼠腹腔接种BCG后，脾DC表面共刺激分子表达较对照组高（P〈0．05）；成年BALB／c小鼠腹腔接种BCG后，脾DC表达CD40、MHC-Ⅱ类分子较对照组高，CD86表达为低（P〈0．05）；新生鼠和成年BALB／c小鼠腹腔接种BCG后，脾DC表面共刺激分子表达差异无明显统计学意义。（3）新生BALB／c小鼠腹腔接种BCG后，脾细胞在体外受到LPS／BCG再次刺激后，较对照组IL-12p70[（68．49±7．78）pg／ml vs（58．64±8．03）pg／ml，P〈0．05]或IL-10[（346．28，280）pg／ml vs（221．90，180）ps／ml，P〈0．05]的分泌较高，IL-6无明显改变。结论BCG接种可通过改变新生BALB／c小鼠脾DC表型和细胞因子分泌诱导脾Th1／Tr1应；BCG接种对成年BALB／c小鼠脾DC的影响作用与新生鼠有所不同。Objective The impact of dendritic cells （DCs） and regulatory T cells （Tr） on the pathogensis of asthma have been investigated over the past decades. As the professional antigen presenting cells, DCs not only prime immune response directing Th0 cells toward different T subtypes but also induce immune tolerance. As an immunoregulator, Bacillus Calmette-Guerin （BCG） has potential to be applied in allergic diseases such as asthma for prevention. Previous study showed that neonatal BCG vaccination could induce Thl/Trl development in mice in vivo. To further identify the mechanism of neonatal BCG vaccination on T cell subsets differentiation, the present study was designed to investigate the impact of BCG vaccination on splenic DCs development in neonatal mice. Methods Neonatal specefic pathogen free （SPF） BALB/c mice （2-3 days） were divided into intraperitonal BCG-treated group, subcutaneous BCG-treated group and control group; simultaneously adult SPF BALB/c mice （6-8 weeks） were divided into intraperitonal BCG- treated and control group. The BCG-treated mice were inoculated with 1 ×10^5 CFU BCG, the mice in control group were not inoculated with any vaccine. Four weeks post BCG vaccination, spleen cells were isolated. With flow eytometry, subtype and maturity of splenic DCs were analysed. Moreover, cells were further separated into mononuclear cell by Ficoll solution. The mononuclear cells were stimulated by 1μg/ml lipopolysaccharidc （LPS） for 18 or 105 CFU /ml BCG for 48 hours at 37 ℃in a humidified atmosphere containing 5% CO2 and cytokines concentration was detected by ELISA. Results （ 1 ） CD11C^＋CD8α^＋and CD11C^＋CD8α^- DCs were found in spleen cells of the BALB/c mice. In comparison with the control group, the percent of CD11C^＋CD8α^- DCs in intraperitoneal BCG group significantly declined（ P 〈0. 01 ） and that of CD11C^＋CD8α^＋ DCs significantly increased （ P 〈 0. 01 ） , there were no significant difference in DC subtypes between intraperitonal and s

关 键 词：婴儿 新生 卡介苗 小鼠 脾 树突状细胞 

分 类 号：R686[医药卫生—骨科学]