摘要
目的 以具有自主知识产权的纳米乳(NEA)和商品化的氢氧化铝[Al(OH)_(3)]作为配方组分,制备出新型NEA/Al(OH)_(3)复合佐剂系统,并对增强MRSA重组亚单位疫苗免疫应答效应的作用进行研究。方法 以血清特异性IgG作为评价指标,将2种乳剂(NEA和MF59)和2种铝盐[Al(OH)_(3)和AlPO4]进行复配筛选;采用多因素实验设计,对初步筛选出的复合佐剂成分进行三因素三水平正交实验,确定优选的NEA/Al(OH)_(3)复合佐剂系统处方;采用金黄色葡萄球菌国际标准株(MRSA252)建立Balb/c小鼠脓毒血症动物感染致死模型,进一步对新型NEA/Al(OH)_(3)复合佐剂系统增强MRSA重组亚单位疫苗抗原HI的免疫应答效应(特异性体液免疫应答、细胞免疫应答、免疫保护效力)进行比较研究,评价其免疫增强的保护效果及初步机制。结果 NEA和Al(OH)_(3)的复配是优选的乳剂与铝盐复合佐剂组分;通过三因素三水平正交实验确定优化的NEA/Al(OH)_(3)复合佐剂系统处方为:HI抗原量60μg、佐剂量200μg、NEA与Al(OH)_(3)的配比为2:8(w/w);脓毒血症动物模型攻毒保护的研究结果表明:基于新型NEA/Al(OH)_(3)复合佐剂系统的MRSA重组亚单位疫苗可于首次免疫后第7天即产生特异性免疫应答,特异性IgG抗体效价最高可达1:143 360,同时诱导出高水平的特异性细胞免疫应答(IL-12P40、INF-γ),LD90的攻毒保护率为80%,具有优于单一佐剂组的良好免疫原性及保护效果。结论 成功优选制备的新型NEA/Al(OH)_(3)复合佐剂系统可有效增强MRSA重组亚单位疫苗的免疫应答水平和保护效力,为新型复合佐剂在重组蛋白疫苗中的应用与开发提供了重要的实验基础。
In this study, a novel NEA/Al(OH)_(3)compound adjuvant system was prepared using nanoemulsion(NEA) with proprietary intellectual property rights and commercial aluminum hydroxide [(AlOH)3] as formula components, and the effect of enhancing the immune response of MRSA recombinant subunit vaccine was studied.Using serum specific IgG as the evaluation index, two emulsions(NEA and MF59) and two aluminum salts [Al(OH)_(3)and AlPO4] were screened. The optimized NEA/Al(OH)_(3)compound adjuvant system prescription was determined by using the multi-factor experimental design, and the compound adjuvant components initially selected were verified by an orthogonal experiment with three factors and threelevels. The international standard strain ofStaphylococcus aureus(MRSA252) was used to establishan animal model of sepsis in Balb/c mice. Furthermore,the enhancing effects of the novel NEA/Al(OH)_(3)compound adjuvant system on immune responses(specific humoral immune response, cellular immuneresponse, immune protective efficacy) of MRSA recombinant subunit vaccine antigen HI was evaluated by comparative study, and the protective effect andpreliminary mechanism of immune enhancement were explored. Data showed that the combination of the NEA andAl(OH)_(3)were the optimal component of emulsion and aluminum salt compound adjuvant. The optimal prescription ofNEA/Al(OH)_(3)compound adjuvant system was determined by three-factor and three-level orthogonal experiment asfollows: HI antigen volume 60 μg, adjuvant volume 200 μg, the ratio of the NEA to Al(OH)_(3)was 2:8(w/w). Thechallenge protection of sepsis animal model showed that the recombinant MRSA subunit vaccine based on the novelNEA/Al(OH)_(3)compound adjuvant system could produce a specific immune response on day 7 after the firstimmunization, with the titer of specific Ig G antibody up to 1:143 360, and induce a high level of specific cellularimmune response(IL-12P40, INF-γ). The challenge protection rate of LD90was 80%, which had betterimmunogenicity and protective effect
作者
曾小强
罗兴
叶演
杨赟
孙红武
曾浩
ZENG Xiaoqiang;LUO Xing;YE Yan;YANG Yun;SUN Hongwu;ZENG Hao(National Engineering Research Center of Immunological Products,Department of Microbiology and Biochemical Pharmacy,College of Pharmacy&Laboratory Medicine,Army Medical University,Chongqing 400038,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2023年第2期176-184,共9页
Immunological Journal
基金
国家自然科学基金(32070924,32000651)
重庆自然科学基金(2014jcyjA0107,2019jcyjA-msxmx0159)
重庆英才计划(CQYC202003220)。
关键词
纳米乳
氢氧化铝
复合佐剂系统
MRSA疫苗
免疫应答
Nanoemulsion
Aluminium hydroxide
Compound adjuvant systems
MRSA vaccine
Immune response
