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目的观察不同剂量霍乱毒素(cholera toxin,CT)联合弓形虫排泄-分泌抗原(excreted-secreted antigens,ESA)滴鼻免疫小鼠的佐剂效应,探索CT作为鼻黏膜佐剂的适宜剂量。方法 5~6周龄BALB/c小鼠60只,随机分为5组,每组12只,分别以0、0.5、1.0、1.5或2.0μg CT联合ESA 20μg滴鼻免疫小鼠,间隔2周进行加强免疫,共2次。末次免疫后30 d,眼静脉丛采血并颈椎脱臼处死小鼠,用ELISA法检测血清IgG和粪便sIgA水平。分离脾、Peyer’s patch(PP)、肠系膜淋巴结(MLN)淋巴细胞。结果 1.5和2.0μg CT组小鼠健康状况下降、存活率降低。免疫后30 d,小鼠粪便sIgA水平随CT剂量的增加而升高,1.0、1.5和2.0μg CT组小鼠粪便sIgA水平显著高于无佐剂组(P<0.05),3组间差异无统计学意义(P>0.05)。CT联合ESA鼻内免疫小鼠后MLN、PP和脾淋巴细胞数显著高于无佐剂组(P<0.05),并均呈现一定的剂量效应,但较高剂量组(1.0、1.5和2.0μg)之间差异无统计学意义(P>0.05)。结论 1.0μg CT联合ESA鼻内免疫小鼠可诱导较高水平的黏膜和系统免疫应答,且对小鼠的健康无不良影响。
Objective To observe the adjuvant effect of different doses of cholera toxin (CT) combined with excreted-secreted antigens (ESA) intranasal immunization mice to explore the appropriate dose of CT as nasal mucosa adjuvant. Methods Sixty BALB / c mice aged 5-6 weeks were randomly divided into 5 groups with 12 mice in each group. Mice were immunized intranasally with 0, 0.5, 1.0, 1.5 or 2.0μg CT and 20μg ESA, To strengthen the immune, a total of 2 times. At 30 days after the last immunization, ophthalmic venous blood was collected and mice were sacrificed by cervical dislocation. Serum IgG and stool sIgA levels were measured by ELISA. Spleen, Peyer’s patch (PP), and mesenteric lymph node (MLN) lymphocytes were isolated. Results The health condition of 1.5 and 2.0μg CT mice decreased and the survival rate decreased. At 30 days after immunization, the sIgA level in feces of mice increased with the increase of CT dose. The levels of sIgA in feces of mice in 1.0, 1.5 and 2.0μg CT groups were significantly higher than those in the non-adjuvant group (P <0.05) Statistical significance (P> 0.05). The numbers of MLN, PP and spleen lymphocytes in mice immunized intranasally with CT combined with ESA were significantly higher than those without adjuvant (P <0.05), and all showed dose effects. However, higher doses of 1.0, 1.5 and 2.0 μg ) Was not statistically different (P> 0.05). Conclusion 1.0μg CT combined with ESA intranasally immunized mice can induce higher levels of mucosal and systemic immune response, and no adverse effects on the health of mice.