基于微针途径的艰难梭菌类毒素A经皮免疫效应研究

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摘要目的比较两种不同免疫方式对艰难梭菌类毒素A 抗体效价的影响，以探究经微针介导艰难梭菌疫苗的经皮免疫效果。方法采用透析培养、纯化、脱毒等方法制备艰难梭菌类毒素A 疫苗后，选择18～22 g 雌性BALB/c 小鼠（SPF 级）共24 只，采用随机数字表法将其分为对照组与观察组，每组各12 只。对照组选用腹腔注射方式，观察组利用微针经皮导入，于处理第0、7、14、28、42 天共分5 次进行免疫，定期检测特异性血清免疫球蛋白G（IgG）和粪便中免疫球蛋白A（IgA）的效价水平变化，最后通过艰难梭菌动物感染实验来评价微针经皮免疫的实际免疫效果。结果两组处理第42 天的血清抗体IgG 效价均高于本组处理第7、14、28 天，差异有统计学意义（P＜0.05）；两组处理第14、28、42 天的血清抗体IgG 效价比较，差异无统计学意义（P＞0.05）。两组处理第42 天的粪便IgA 效价高于本组处理第7、14、28 天，差异有统计学意义（P＜0.05）；两组处理第7 天的粪便IgA 效价比较，差异无统计学意义（P＞0.05）；观察组处理第14、28、42 天的粪便IgA 效价高于对照组，差异有统计学意义（P＜0.05）。攻毒后两组小鼠均无死亡现象，经皮免疫效果良好。处死小鼠后，不同于未经免疫的小鼠攻毒后表现出的黏膜上皮溃疡及严重黏膜炎症，两组小鼠的炎症反应均较轻。结论微针途径介导艰难梭菌类毒素A 疫苗具有较好的黏膜免疫效果。

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关键词 ：艰难梭菌类毒素, 疫苗, 微针, 艰难梭菌感染

Abstract：Objective The effect of two different immunization methods on the titer of Clostridium difficile toxoid A were compared to explore the transdermal immune effect of microneedle-mediated Clostridium difficile vaccine.Meth-ods After preparing Clostridium difficile toxoid A vaccine by dialysis,culture,purification,detoxification,twenty-four 18-22 g female BALB/c mice(SPF grade)were selected.They were divided into control group and observation group by random number table method,with 12 mice in each group.The control group was given intraperitoneal injection,and the observation group was immunized with microneedles percutaneously.At the 0,7th,14th,28th and 42th day of treatment,the titer levels of specific serum immunoglobulin G(IgG)and fecal secretory immunoglobulin A(IgA)were regularly detected.Finally,the actual immune effect of microneedle percutaneous immunization was evaluated by Clostridium difficile animal infection experiment.Results The serum antibody IgG titer of the two groups at the 42th day of treatment was higher than that at the 7th,14th and 28th day of treatment,and the difference was statistically significant(P＜0.05).There was no significant difference in the titer of serum antibody IgG between the two groups at the 14th,28th and 42th day of treatment(P＞0.05).The fecal IgA titer of the two groups at the 42th day of treatment was higher than that at the 7th,14th and 28th day of treatment,and the difference was statistically significant(P＜0.05).There was no significant difference in fecal IgA titer between the two groups at the 7th day(P＞0.05).The fecal IgA titer of the observation group at the 14th,28th and 42th day of treatment was higher than that of the control group in the same period,and the difference was statistically significant(P＜0.05).There was no death in both groups after challenge,and the percutaneous immune effect was good.After the mice were killed,compared with the mucosal epithelial ulcer and severe mucosal inflammation in the unimmunized mice,the inflammatory reactions in both groups were mild.Conclusion Microneedle-mediated Clostridium difficile toxin A vaccine has good mucosal immune effect.

Key words： Clostridium difficile toxoid Vaccine Microneedle Clostridium difficile infection

收稿日期: 2021-06-17

基金资助:西北民族大学国家级大学生创新创业训练计划项目（202010742115）；国家自然科学基金资助项目（8156 0090）

引用本文:

何菁荣；张祎；代文秀；王月颖；傅思武. 基于微针途径的艰难梭菌类毒素A经皮免疫效应研究[J]. 中国当代医药, 2022, 29(11): 40-44.
HE Jingrong ZHANG Yi DAI Wenxiu WANG Yueying FU Siwu▲. Study on percutaneous immune effect of Clostridium difficile toxoid A mediated by microneedle. 中国当代医药, 2022, 29(11): 40-44.

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