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目的:观察用白细胞介素 2(interleukin-2,IL-2)基因修饰小鼠骨髓来源的树突状细胞(dendric cells, DC)后,DC表达细胞因子和趋化因子的变化及DC体内免疫后,小鼠体内主要脏器中IL-2表达情况。探讨其对抗原提呈微环境的改善及其可能的机制。方法:用EIJSA方法检测DC培养上清和小鼠血清及脏器中IL-2的含量,用RT-PCR方法检测DC中IL-2,IFN-γ,IL-12,GM-CSF,IL-4,IL-10,TNF-α,IL-1-β,MIP-1β,MCP-1,RANTES和IP10等的表达,用FACS分析DC表面IL-2受体的变化。结果:经IL-2基因修饰后,DC除了分泌高水平的IL-2外,尚能分泌一定水平的IFN-γ和IL-12,其表面表达的IL-2Rα密度增加。RT-PCR分析表明其表达多种细胞因子和趋化因子,用IL-2基因修饰的DC免疫小鼠后,血清和脏器中能检测到高水平的IL-2。结论:IL-2基因修饰DC,能促进DC表达IL-2受体,并以自分泌形式分泌高水平的ILZ,优化其抗原提呈的微环境,提示可增强DC的抗原提呈功能。
Objective: To observe the changes of cytokines and chemokines expressed in DCs and DC in vivo after modification of mouse bone marrow-derived dendritic cells (DCs) with interleukin-2 (IL-2) gene Afterwards, IL-2 expression was observed in major organs in mice. Explore its improvement of the microenvironment of antigen presentation and its possible mechanisms. Methods: The content of IL-2 in DC culture supernatant and mouse serum and organ was detected by EIJSA method. IL-2, IFN-γ, IL-12, GM-CSF and IL-4 in DC were detected by RT-PCR. Expression of IL-2, IL-10, TNF-α, IL-1-β, MIP-1β, MCP-1, RANTES and IP10, etc. The changes of IL-2 receptor on DC surface were analyzed by FACS. RESULTS: After IL-2 gene modification, DCs secreted high levels of IL-2, but secreted certain levels of IFN-γ and IL-12, and the surface expression of IL-2Rα increased. RT-PCR analysis showed that it expresses a variety of cytokines and chemokines. After immunization of mice with IL-2 gene-modified DCs, high levels of IL-2 can be detected in serum and organs. CONCLUSION: DCs modified by IL-2 gene can promote the expression of IL-2 receptors in DCs, secrete high levels of ILZ in autocrine form, and optimize the microenvironment for antigen presentation, suggesting that DCs can enhance the antigen presentation function of DCs.