壳聚糖佐剂白喉类毒素疫苗对小鼠的免疫效果评价  

Evaluation of immune effect of diphtheria toxoid vaccine with chitosan as adjuvant on mice

作　　者：李晓彤 张晨宵 于广福 沈书园 赵英会 邵祝军[2] 郭蓬勃 LI Xiaotong;ZHANG Chenxiao;YU Guangfu;SHEN Shuyuan;ZHAO Yinghui;SHAO Zhujun;GUO Pengbo(School of Clinical and Basic Medicine(lnstitute of Basic Medicine),Shandong First Medical University&Shandong Academy of Medical Sciences,Jinan 250000,Shandong Province,China;不详)

机构地区：[1]山东第一医科大学(山东省医学科学院)临床与基础医学院(基础医学研究所),山东济南250000 [2]中国疾病预防控制中心传染病预防控制所,北京102206

出　　处：《中国生物制品学杂志》2023年第11期1286-1290,共5页Chinese Journal of Biologicals

基　　金：山东省自然科学基金青年项目(ZR2022QH039);山东省大学生创新创业训练项目(S202110439081);山东第一医科大学2020年人才专项21科教融合(921001007002)。

摘　　要：目的评价以壳聚糖为佐剂的白喉类毒素(diphtheria toxoid,DT)疫苗对小鼠的免疫效果,为DT新型佐剂疫苗的制备提供实验依据。方法将30只雄性C57BL/6小鼠分为黏膜免疫组和体液免疫组,每组再随机分为6组:阴性对照组(PBS)、阳性对照组(DT)、铝佐剂组、壳聚糖佐剂组(配成含壳聚糖0.5%的终溶液)、铝佐剂疫苗组和壳聚糖佐剂疫苗组,每组5只。黏膜免疫组和体液免疫组分别通过滴鼻和腹腔注射接种1次,PBS缓冲液500μL/只,DT10μg/只,铝佐剂500μg/只,壳聚糖佐剂500μL/只。观察小鼠状态,并分别于接种前(0 d)、接种后7、21、35 d采集小鼠眼球血清,同时收集鼻洗液,采用ELISA法分别检测血清IgG抗体水平和鼻洗液sIgA抗体水平。结果小鼠腹腔注射及鼻滴壳聚糖佐剂疫苗后,精神及行为状态正常。壳聚糖佐剂疫苗可有效诱导小鼠产生白喉IgG抗体,与经典铝佐剂疫苗组相比,免疫后35 d差异无统计学意义(F=0.742<F_(0.05(1,8)),P>0.05);也可有效诱导小鼠产生sIgA,免疫后21 d明显高于经典铝佐剂疫苗组(F=127.926>F_(0.05(1,8)),P<0.05)。结论壳聚糖佐剂DT疫苗安全性良好,可有效诱导小鼠黏膜免疫和体液免疫,具有替代铝佐剂制备疫苗的潜力,且可考虑制备鼻滴疫苗。Objective To evaluate the immune effect of the diphtheria toxoid(DT)vaccine using chitosan as adjuvant on mice,so as to provide an experimental basis for the preparation of novel adjuvant vaccines of DT.Methods A total of 30 male C57BL/6 mice were divided into mucosal immunization group and humoral immunization group,and each group was randomly divided into 6 groups:negative control group(PBS),positive control group(DT),aluminum adjuvant group,chitosan adjuvant group(final solution with 0.5%chitosan),aluminum adjuvant vaccine group and chitosan adjuvant vaccine group,with 5 mice in each group.Mucosal immunization group and humoral immunization group were inoculated with 500μL PBS buffer,10μg DT,500μg aluminum adjuvant and 500μL chitosan adjuvant per mouse by nasal drip and intraperitoneal injection respectively.The mice were observed for the status,collected for the eyeball serum before inoculation(0 d)and 7,21,35 d after inoculation,and collected for the nasal lotion simultaneously.The levels of IgG antibody in serum and slgA antibody in nasal lotion were detected by ELISA.Results After intraperitoneal injection and nasal drip of chitosan adjuvant vaccine,the mental and behavioral state of mice was normal.Chitosan adjuvant vaccine effectively induced the IgG antibody against diphtheria in mice with not significant difference,compared with the classic aluminum adjuvant vaccine group(F=127.926>F_(0.05(1.8)),P>0.05);It also induced the production of slgA in mice,which was significantly higher than that in the classical aluminum adjuvant vaccine group 21 d after immunization(F=127.926>F_(0.05(1.8)),P<0.05).Conclusion Chitosan adjuvant DT vaccine can ffectively stimulate the mucosal immunity and humoral immunity in mice with good safety,and has the potential to be an alternative to aluminum adjuvant for vaccine preparation,which can be considered as nasal drop vacines.

关 键 词：壳聚糖 白喉类毒素 疫苗 免疫效果 

分 类 号：R18[医药卫生—流行病学]