蛋白体佐剂及用于鼠疫F1-Ⅴ抗原的免疫效果

目的以蛋白体(proteosomes)佐剂,非共价结合鼠疫F1-V重组蛋白为免疫原,探讨滴鼻免疫BALB/c小鼠后诱导的免疫应答和免疫保护效果。方法佐剂与鼠疫F1-V重组蛋白为免疫原非共价结合,滴鼻免疫BALB/c小鼠4次后,采用间接ELISA检测血清特异性抗F1-V的IgG和IgA抗体及抗体亚型分类,并检测鼻咽、肺、小肠及阴道灌洗液中特异性抗F1-V的黏液分泌型IgA;并用流式细胞术检测鼻相关淋巴组织淋巴细胞、脾淋巴细胞、肠系膜淋巴结及小肠PP结T淋巴细胞表型的变化。第4次免疫后7d,用100 LD_(50)的鼠疫141强毒株进行腹腔攻毒。结果(1)以蛋白体为佐剂的鼠疫F1-V抗原与单纯的F1-V组相比,蛋白体疫苗组诱导血清IgG、IgA抗体显著升高(P<0.01),同时蛋白体疫苗组能诱导鼻咽、肺、小肠和阴道内多个黏膜部位特异性IgA抗体的产生,尤其是肺和生殖道冲冼液内抗体升高极为显著(P<0.01);(2)蛋白体疫苗组主要引起IgG1型抗体,主要诱导T_H2型免疫反应;(3)蛋白体疫苗组NALT和SP中CD4~ /CD8~ 比值比PBS对照有显著增高(P<0.01),MLN和PP中CD4~ /CD8~ 比值与PBS对照差异无统计学意义(P>0.05)。(4)小鼠在100 LD_(50)的鼠疫141强毒株腹腔攻毒后鼠疫F1-V重组蛋白组小鼠免疫保护率为0,而蛋白体佐剂疫苗组小鼠免疫保护率为67%。结论以自制的蛋白体为鼠疫F1-V抗原的佐剂滴鼻免疫小鼠,蛋白体不仅提高鼠疫F1-V抗原的系统免疫应答,而且能诱导小鼠呼吸道、消化道和生殖道局部黏膜免疫应答。蛋白体佐剂鼠疫疫苗对100 LD_(50)的鼠疫141强毒株腹腔攻毒具有一定的免疫保护作用,这为鼠疫黏膜疫苗的研制提供候选材料,也为鼠疫黏膜疫苗深入研究奠定了基础。

Proteosome adjuvant and its application in anti-plague immunity induced by recombinant F1-Ⅴ protein

Objective To develop outer membrane protein proteosomes from Neisseria meningitids as an adjuvant for mucnsal and systemic immunity elicited by intranasal immunization with F1-V vaccine.Methods Outer membrane protein proteosomes were prepared from group B serotype 2b Neisseria meningitids and proteosome- adjuvanted F1-V vaccine was prepared by formulating recombinant F1-V protein with proteosome protein at an ap- propriate ratio.After intranasal immunization of mice with proteosome-adjuvanted F1-V vaccine,mucosal and sys- temic immunity was determined by ELISA,FACS and experimental challenge.Results Intranasal immunization of mice with proteosome-adjuvanted F1-V vaccine induced significant antibody response.67% mice immunized in- tranasally with proteosome-adjuvanted F1-V vaccine were protected against challenge by 100 LD_(50)of Y.pestis(141 strain).Conclusion The proteosome adjuvant/delivery system is capable of interacting not-covalently with anti- gents to form vaccines which,elicited enhanced immunity and so is a potential vaccine for clinical use.