摘要
目的 研究重组大肠杆菌疫苗E.coli LLO/OVA刺激小鼠骨髓树突状细胞的细胞因子表达,探讨该疫苗对BM-Dc的免疫刺激作用.方法 以E.coli OVA为对照,采用基因芯片、RT-PCR和ELISA在基因和蛋白质水平检测E.coli LLO/OVA刺激C57BL/6小鼠骨髓树突状细胞细胞因子表达情况.结果 经E.coli LLO/OVA刺激后4～24 h,BMDC出现一系列细胞因子基因表达上调,尤其在刺激后4～8 h BMDC的细胞因子基因上调表达明显,其中G-CSF和IFN-γ的表达明显高于E.coli OVA刺激后的BMDC RT-PCR检测也证实E.coli LLO/OVA刺激8 h后BMDC的IFN-γmRNA转录水平明显高于E.coli OVA刺激的BMDC FLISA结果显示E.coli LLO/OVA刺激BMDC后12～24 h,培养上清液内IFN-γ的含量明显高于E.coli OVA刺激后的BMDC,而IL-10的含量则明显降低.结论 E.coli LLO/OVA刺激小鼠BMDC上调表达一系列细胞因子,并且较E.coli OVA刺激的BMDC更明显上调G-CSF和IFN-γ基因表达.ILO作为重组疫苗E.coli OVA的免疫佐剂在刺激BMDC表达促进机体免疫的细胞因子中发挥重要作用.
Objective To investigate the immunostimulating effects of recombinant E. coli LLO/OVA on C57BL/6 murine bone marrow dendritic cells （BMDC） by detecting cytokine expressions of the cells. Methods Microarray, reverse transefiption-polymerase chain reaction （RT-PCR）, and enzyme-linked immunosorbent assay （ELISA） were used to detect the cytokine gene transcription and protein expressions of murine BMDC stimulated by E. coli LLO/OVA and E. coli OVA. Results Compared with OVA-stimulated BMDC, the expressions of cytokines in E. coli LLO/OVA-stimulated BMDC was up-regulated, especially the expressions of colony stimulating factor 3 （G-CSF） and IFN-γ, but the expression of IL- 10 was decreased. Conclusion Recombinant E. coli LLO/OVA can effectively enhance the immune function of BMDC by up-regulating series of cytokines, especially up-regulating IFN-γ and G-CSF. LLO is an effective adjuvant in recombinant E. coli vector in promo-ting immune activity of murine BMDC.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2007年第4期402-405,408,共5页
Immunological Journal
