细粒棘球绦虫重组抗原铁蛋白诱导小鼠骨髓树突状细胞免疫应答的机制研究

目的 探索细粒棘球绦虫重组抗原铁蛋白(rEg.ferritin)免疫小鼠产生抵抗原头蚴感染的保护机制。方法 利用rEg.ferritin和囊液（HF）两种细粒棘球绦虫抗原与小鼠骨髓树突状细胞(BMDC)进行体外培养，共分6组:HF组（加入HF 30 μg/mL）、rEg.ferritin组（加入抗原rEg.ferritin 1 μg/mL）均为单独刺激组，rEg.ferritin+LPS 组（加入抗原rEg.ferritin 1 μg/mL和LPS 100 ng/mL）和HF+LPS组（加入HF 30 μg/mL和 LPS 100 ng/mL）为联合刺激组，LPS阳性对照组（加入LPS 100 ng/mL），空白对照组（不加任何抗原）。通过电镜检测各组BMDC形态学的变化；流式细胞技术检测各组DC的表面分子表达、吞噬能力及诱导T细胞增殖能力的变化；酶联免疫吸附试验(ELISA) 检测各组BMDC培养上清液中所含Th1型〔白介素（IL）-12p70，肿瘤坏死因子-α（TNF-α）〕和Th2型（IL-6和IL-10）细胞因子含量。结果 rEg.ferritin 组和rEg.ferritin+LPS组BMDC具有成熟的形态，表面突起数目高于空白对照组、HF组和HF+LPS组（P<0.05）；高表达MHCⅡ、CD80、CD86及CD40分子，与空白对照组比较，差异有统计学意义（ P<0.05)；诱导T细胞增殖能力强于空白对照组、HF组和HF+LPS组（P<0.05），吞噬能力弱于HF组；高表达IL-6、IL-12p70、TNF-α及IL-10，与空白对照组比较，差异有统计学意义（ P<0.05)。结论 rEg.ferritin可以刺激BMDC细胞成熟，促进T增殖，释放高含量的Th1型及Th2型细胞因子，激活Th1型或Th2型细胞，引发免疫应答。

Effect of Recombinant Antigen Ferritin on DC Induce Immune Response

Objective To explore the immunoreaction against protoscolex infection by the recombinant ferritin of echinococcus granulosus （rEg.ferritin） immunized mice. MethodsBone marrow deriedstem cells (BMDCs) were collected and exposed in different antigens by cell culture and included 6 groups, there were hydatid fluid （HF） group (HF 30 μg/mL), rEg.ferritin group (rEg.ferritin 1 μg/mL)，rEg.ferritin+LPS group (rEg.ferritin 1 μg/mL and LPS 100 ng/mL), HF+LPS group (HF 30 μg/mL and LPS 100 ng/mL), LPS group (LPS 100 ng/mL) and control (not add antigen). Morphology of BMDCs in different groups were detected by scan electron microscope (SEM). Phagocytic-ability, T cell multiplication capability and surface marker expression of BMDCs in different groups were detected by flow cytometry.Cytokines of Th1 〔interleukin (IL)-12p70, tumor necrosis factor-α (TNF-α)〕 and Th2 (IL-6 and IL-10) in surpernatant of BMDCs in different groups were detected by enzyme-linked immunosorbent assay (ELISA). Results rEg.ferritin or rEg.ferritin+LPS stimulated BMDCs maturation，which the dendrites number of BMDCs in these 2 groups were more and longer than control , HF and HF+LPS groups ( P<0.05)，and induced higher levels of surface molecules major histocompatibility complexⅡ (MHCⅡ), CD80, CD86 and CD40, also had strong ability to induce T cell multiplication than control, HF, HF+LPS groups ( P<0.05), but had weak phagocytosis than HF group, and secreted higher levels of IL-6, IL-12p70, TNF-α and IL-10 than control group. ConclusionBMDCs are stimulated with rEg.ferritin and became maturation. By activating T cell and releasing more cytokines of Th1 and Th2, rEg.ferrtin can induce BMDCs to produce Th1 and Th2 immunoreaction.