不同佐剂细胞基质四价流感裂解疫苗的小鼠免疫效果评价  

Immunogenicity of MDCK cell-based quadrivalent influenza split vaccine with different adjuvants in mice

作　　者：年悬悬 刘京 邓涛 吕传硕 张青梅 彭飞霞 张家友 杨晓明 Nian Xuanxuan;Liu Jing;Deng Tao;Lyu Chuanshuo;Zhang Qingmei;Peng Feixia;Zhang Jiayou;Yang Xiaoming(National Engineering Technology Research Center for Combined Vaccines,Wuhan 430207,China;The Second Research Department of Viral Vaccine,Wuhan Institute of Biological Products Co.,Ltd.,Wuhan 430207,China;China National Biotech Group Company Limited,Beijing 100024,China)

机构地区：[1]国家联合疫苗工程技术研究中心,武汉430207 [2]武汉生物制品研究所有限责任公司病毒性疫苗研究二室,430207 [3]中国生物技术股份有限责任公司,北京100024

出　　处：《中华微生物学和免疫学杂志》2021年第9期711-718,共8页Chinese Journal of Microbiology and Immunology

基　　金："重大新药创制"科技重大专项支持(2016ZX09106003-008)。

摘　　要：目的评价犬肾细胞基质四价流感裂解疫苗(MDCK-Va)配伍不同佐剂后的免疫原性。方法将高、中、低剂量的MDCK-Va和鸡胚基质四价流感裂解疫苗(egg-Va)原液分别肌肉免疫BALB/c小鼠2次,间隔3周,小鼠眼静脉采血,分离血清,血凝抑制(hemagglutination inhibition,HI)试验检测抗体效价。不同剂量的MDCK-Va分别配伍QS21、AddVax、PolyI∶C、CpG ODN 1826以及AddVax/PolyI∶C(Add/Poly)联合佐剂,免疫小鼠,初次免疫21 d后,小鼠眼静脉采血,HI和微量中和(microneutralization,MN)试验检测抗体效价,同等剂量加强免疫21 d后,再次采血检测抗体效价。10μg MDCK-Va配伍上述佐剂,加强免疫5 d后,取小鼠脾脏检测脾指数和脾细胞分型。结果MDCK-Va诱导的各型别(H1N1、H3N2、B/Victoria和B/Yamagate)的血清效价均非劣效egg-Va诱导的血清效价。QS21/Va、AddVax/Va、PolyI∶C/Va、CpG ODN 1826/Va,以及Add/Poly/Va诱导的小鼠血清HI抗体效价均高于MDCK-Va,尤其是QS21/Va和Add/Poly/Va诱导的保护性效价,其差异具有统计学意义。对于H1N1疫苗,血清HI抗体和MN抗体之间的Pearson相关系数为0.737~0.910;对于H3N2亚型,HI抗体和MN抗体的Pearson相关系数为0.839~0.947。QS21/Va组具有明显的脾指数增高,但淋巴细胞占比降低。QS21/Va和Add/Poly/Va对小鼠脾脏中性粒细胞和CD4/CD8细胞的刺激明显强于其他佐剂。结论Add/Poly/Va组对MDCK-Va具有较强的免疫增强作用,可作为MDCK-Va的候选佐剂。HI和MN试验检测的抗体具有很强的正相关性。Objective To evaluate the immunogenicity of Madin-Darby canine kidney(MDCK)cell-based quadrivalent influenza split vaccine(MDCK-Va)combined with different adjuvants.Methods Different doses of MDCK-Va and chicken embryo-based quadrivalent influenza split vaccine(egg-Va)were intramuscularly immunized BALB/c mice twice with an interval of three weeks.Serum samples were collected to detect antibody titers using hemagglutination inhibition(HI)assay.BALB/c mice were immunized with different doses of MDCK-Va combined with QS21,AddVax,PolyI∶C,CpG ODN 1826 and AddVax/PolyI∶C(Add/Poly),respectively.HI and microneutralization assays were used to detect antibody titers 21 d after the first and booster immunization.Spleen tissues were collected from the mice immunized with 10μg MDCK-Va combined with the above adjuvants 5 d after the booster immunization to analyze spleen index and the types of spleen cells.Results The immunoprotective effect of MDCK-Va was not inferior to that of egg-Va.MDCK-Va combined with each of the above adjuvants could induce higher HI antibody titer than MDCK-Va alone,especially the QS21/Va and Add/Poly/Va groups,and the differences were statistically significant.For H1N1 vaccine,the Pearson′s correlation coefficient(r)between HI antibody and neutralizing antibody was 0.737-0.910,and for H3N2 subtype vaccine,the value of r was 0.839-0.947.Compared with the MDCK-Va group,the QS21/Va group showed significantly increased spleen index and decreased proportion of single lymphocytes.QS21 and Add/Poly were much better than other adjuvants in stimulating mouse splenic neutrophils and CD4/CD8 cells.Conclusions Add/Poly had a stronger immune enhancement effect on MDCK-Va,suggesting that it was a potential adjuvant for MDCK-Va.The antibody titer detected by HI and MN assays had a strong positive correlation.

关 键 词：细胞基质四价流感裂解疫苗 佐剂 稳定性 免疫效果 

分 类 号：R392[医药卫生—免疫学]