流感病毒灭活疫苗可溶性微针制备及评价

发布时间：2018-07-12 22:01

本文选题：微针 + 可溶性　；参考：《免疫学杂志》2017年06期

【摘要】：目的初步研究负载流感全病毒灭活疫苗可溶性微针体外评价方法和经皮免疫效果。方法将流感全病毒灭活疫苗溶液与微针基质材料混匀干燥后复溶,体外血凝试验检测疫苗血凝素活性,透射电镜和动态光散射分析疫苗微针复溶后流感全病毒疫苗变化情况;选择筛选的材料羧甲基纤维素和硫酸软骨素作为主要基质与疫苗混匀制备微针溶液,滴加到微针制作模具上,抽真空使疫苗基质溶液填充针体,室温干燥、脱模,贴于猪皮肤检测微针硬度,将Cy7标记流感疫苗可溶性微针贴于小鼠腹部后活体成像检测流感疫苗皮内存留时间,动物免疫试验第2次免疫后14 d检测血抑效价。结果通过血凝试验筛选出硫酸软骨素和羧甲基纤维素为主要基质,贴片疫苗血凝活性为96%和86%,显著高于其它3种材料贴片疫苗活性;透射电镜显示PBS对照组中流感灭活病毒颗粒多呈单个散在分布,罕见聚团,2种贴片复溶后流感灭活病毒多呈聚团,罕见单个病毒颗粒;动态光散射分析显示疫苗贴片复溶后病毒颗粒粒径明显增大,高于对照组中病毒粒径;皮肤组织病理显示2种疫苗微针均可刺穿猪皮肤;活体成像显示疫苗微针刺穿小鼠皮肤并溶解于皮内,疫苗存留时间较肌肉长;Balb/c小鼠免疫后分离血清,血凝抑制试验显示2种微针均产生血抑效价且硫酸软骨素微针血抑效价高于羧甲基纤维素微针。结论可溶性疫苗微针可刺穿皮肤溶于皮内产生免疫应答,为微针疫苗的研究提供了思路。
[Abstract]:Objective to study the in vitro evaluation and percutaneous immunization of inactivated influenza virus vaccine with soluble microneedle. Methods Influenza virus inactivated vaccine solution was mixed with microneedle matrix and then redissolved. Hemagglutination test in vitro was used to detect hemagglutinin activity. Transmission electron microscopy and dynamic light scattering were used to analyze the changes of influenza virus vaccine after microneedle relysis. The selected materials, carboxymethyl cellulose and chondroitin sulfate, were used as the main matrix to prepare the microneedle solution, and the microneedle solution was added to the microneedle mould. The vaccine matrix solution was filled with the needle body, dried at room temperature, and demoulded. The microneedle hardness of swine skin was detected, the soluble microneedle of Cy7 labeled influenza vaccine was applied to mouse abdomen to detect the survival time of influenza vaccine skin, and the blood inhibitory value was detected 14 days after the second immunization of animal immunity test. Results Chondroitin sulfate and carboxymethyl cellulose were selected as the main matrix by hemagglutination test. The hemagglutination activity of patch vaccine was 96% and 86%, which was significantly higher than that of the other three materials. Transmission electron microscopy showed that influenza inactivated virus particles in PBS control group were mostly distributed in a single form. Dynamic light scattering (DLS) analysis showed that the particle size of the virus increased significantly after redissolution of the vaccine patch, which was higher than that of the control group, and the skin histopathology showed that two kinds of vaccine microneedles could puncture the pig skin. In vivo imaging showed that the vaccine punctured the skin and dissolved in the skin of the mice, and the vaccine remained longer than the muscle and separated the serum from Balb / c mice after immunization. The results of hemagglutination inhibition test showed that both microneedles produced blood inhibitory potency and the blood inhibitory effect of chondroitin sulfate microneedles was higher than that of carboxymethyl cellulose microneedles. Conclusion soluble vaccine microneedle can penetrate the skin and produce immune response in skin, which provides a way for the study of microneedle vaccine.
【作者单位】：宁夏医科大学基础医学院病原生物学与免疫学教研室;军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室;中国科学院理化技术研究所;
【基金】：国家科技重大专项课题(2013ZX10004003001)
【分类号】：R392-33

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