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目的研究细粒棘球绦虫(Eg)重组双歧杆菌(Bb)-Eg95-EgA31融合蛋白免疫小鼠后诱导的免疫应答。方法 56只SPF级雌性Balb/c小鼠随机均分7组,分别为重组Bb-Eg95-EgA31皮下注射组(A组)、肌肉注射组(B组)、鼻腔内接种组(C组)、口服灌胃组(D组)、空载体对照组(E组)、Bb对照组(F组)和Bb培养用液体培养基(MRS)对照组(G组)。免疫后8周各组鼠用50个Eg原头节攻击,攻击25周后,处死小鼠,分离细粒棘球蚴包囊并称重,计算囊重抑制率;ELISA检测血清IgG及其亚类和IgE和脾细胞上清液IL-12、IFN-γ、TNF-α和IL-10水平;MTT法测定脾细胞增殖反应;流式细胞仪检测脾CD4+和CD8+T细胞百分比和脾细胞凋亡发生率。结果重组Bb-Eg95-EgA31免疫组小鼠的囊重抑制率分别为45.33%、41.33%、70.67%和62.67%;血清IgG、IgG2a、IgG2b和IgG1水平升高,IgG3和IgE降低;脾IFN-γ、IL-12和TNF-α水平升高,IL-10水平降低;脾T淋巴细胞明显增殖;脾CD4+和CD8+T细胞显著增加;脾细胞凋亡发生率显著降低。结论细粒棘球绦虫重组Bb-Eg95-E-gA31融合蛋白能诱导小鼠产生有效的保护性免疫应答。
Objective To study the immune response induced by recombinant Echinococcus granulosus (Eg) recombinant Bb-Eg95-EgA31 fusion protein. Methods 56 SPF female Balb / c mice were randomly divided into 7 groups: subcutaneous injection of recombinant Bb-Eg95-EgA31 (group A), intramuscular injection of group B (intranasal injection), group C (Group D), empty vector control group (group E), Bb control group (group F) and Bb culture liquid medium (group MR) control group (group G). Eight weeks after the immunization, the mice in each group were challenged with 50 Eg protoscoleces. After being attacked for 25 weeks, the mice were sacrificed, cystic echinococcosis was isolated and weighed, and the inhibition rate of cystic weight was calculated. The serum IgG and its subunit were detected by ELISA The levels of IL-12, IFN-γ, TNF-α and IL-10 in the supernatant of IgE and spleen cells were measured. The proliferation of spleen cells was determined by MTT assay. The percentage of splenic CD4 + and CD8 + T cells and the percentage of splenocytes were detected by flow cytometry The incidence of apoptosis. Results The inhibition rates of capsule weight in recombinant Bb-Eg95-EgA31 immunized mice were 45.33%, 41.33%, 70.67% and 62.67%, respectively. Serum IgG, IgG2a, IgG2b and IgG1 levels were elevated, IgG3 and IgE were decreased, γ, IL-12 and TNF-α levels, IL-10 levels decreased; splenic T lymphocytes significantly proliferated; splenic CD4 + and CD8 + T cells were significantly increased; and the incidence of splenocyte apoptosis was significantly decreased. Conclusion Echinococcus granulosus recombinant Bb-Eg95-E-gA31 fusion protein can induce effective protective immune response in mice.