EV-A71灭活疫苗长期稳定性考察及结果分析
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摘要
目的考察肠道病毒A组71型(EV-A71)灭活疫苗的长期稳定性。方法对中国医学科学院医学生物学研究所生产的,在2~8℃条件下保存54个月的Ⅲ期临床样品,进行全面检定以分析各质量指标的变化;用原倍疫苗样品以0、28d两剂腹腔免疫6~8周龄的Balb/c小鼠,检测其血清抗体效价,进行疫苗效力单位(EU)测定;用2倍系列稀释法稀释疫苗,取不同稀释度样品免疫6~8周龄的Balb/c小鼠,检测其血清抗体效价,进行半数有效量ED50测定。结果该样品在2~8℃保存54个月,所有指标均在质量标准的合格要求范围内,仅pH值7.4接近标准上限;抗原含量为180 U/ml,与原检测值相比,下降仅9.1%;半数有效量(ED50)为6.29U,与原检测结果4.90U保持了良好一致;疫苗初次免疫小鼠后28d抗体阳转率为100%,中和抗体效价为1∶16~1∶256,GMT值为1∶52,EU值为6.5;加强免疫后第7天,中和抗体效价为1∶1 024~1∶4 096,几何平均滴度GMT大幅度上升至1∶1 552.1,其EU值为194.0;疫苗样品稀释至45 U/ml的抗原量,仍能使免疫小鼠产生100%的抗体阳转,且GMT值达1∶27.9。结论 EV-A71灭活疫苗于2~8℃条件下保存54个月,其质量指标无明显变化。
Objective To study the long-term stability of the EV-A71 inactivated vaccine. Methods A comprehensive test of EV-A71 inactivated vaccine,which was used for phaseⅢ clinical trial and stored at 2-8 ℃for 54 months,is performed to analyze the changes of quality indicators.Balb/c mice of 6-8weeks were immunized with the vaccine at 0and 28 days by intraperitoneal injection;the serum antibody titer and EU were detected;Balb/c mice were immunized with two fold serial dilution vaccine,and ED50 was evaluated. ResultsThis batch of vaccine was stored at 2-8 ℃ for 54 months,and all indicators reach the requirement of quality standard;antigen concentration(180 U/ml)only decreased 9.1% comparing to the original level;ED50(6.29U)is consistent with its original level(4.90 U).The positive seroconversion rate is 100% at 28 day point after primary immunization,and neutralization antibody titer 1∶16-1∶256,GMT 1∶52,EU6.5.Neutralization antibody titer reached 1∶1 024-1∶4 096 at 7day point after booster immunization,GMT increased to 1∶1 552.1,and EU is 194.0.The positive seroconversion rate also reached 100% when the vaccine was diluted to 45U/ml,and GMT was 1∶27.9. Conclusions After storing at 2-8 ℃ for 54 months,the EV-A71 inactivated vaccine showed no obvious changes in quality indicators.
Objective To study the long-term stability of the EV-A71 inactivated vaccine. Methods A comprehensive test of EV-A71 inactivated vaccine,which was used for phaseⅢ clinical trial and stored at 2-8 ℃for 54 months,is performed to analyze the changes of quality indicators.Balb/c mice of 6-8weeks were immunized with the vaccine at 0and 28 days by intraperitoneal injection;the serum antibody titer and EU were detected;Balb/c mice were immunized with two fold serial dilution vaccine,and ED50 was evaluated. ResultsThis batch of vaccine was stored at 2-8 ℃ for 54 months,and all indicators reach the requirement of quality standard;antigen concentration(180 U/ml)only decreased 9.1% comparing to the original level;ED50(6.29U)is consistent with its original level(4.90 U).The positive seroconversion rate is 100% at 28 day point after primary immunization,and neutralization antibody titer 1∶16-1∶256,GMT 1∶52,EU6.5.Neutralization antibody titer reached 1∶1 024-1∶4 096 at 7day point after booster immunization,GMT increased to 1∶1 552.1,and EU is 194.0.The positive seroconversion rate also reached 100% when the vaccine was diluted to 45U/ml,and GMT was 1∶27.9. Conclusions After storing at 2-8 ℃ for 54 months,the EV-A71 inactivated vaccine showed no obvious changes in quality indicators.
引文
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[2]Yang JJ,Li W,Wang YL.Research progress of enterovirus71and its vaccine[J].Weishengwuxue Mianyixue Jinzhan,2011,39(2):51-56.(in Chinese)杨俊杰,李薇,王玉琳.肠道病毒71型及其疫苗的研究进展[J].微生物学免疫学进展,2011,39(2):51-56.
[3]Zhou HL,Li HD.Clinical analysis on humoral immunity in children with hand-foot-mouth disease by EV71infection[J].Zhonghua Shiyan He Linchuang Ganranbing Zazhi(Dianzi Ban),2011,5(2):208-211.(in Chinese)周海兰,李华东.EV71感染的手足口病患儿体液免疫临床分析[J].中华实验和临床感染病杂志(电子版),2011,5(2):208-211.
[4]Xu HM,Lai FF.Effect of enterovirus 71on nervous system and immune function and research progress of vaccine[J].Shiyong Erke Linchuang Zazhi,2012,27(22):1701-1704.(in Chinese)许红梅,赖方方.肠道病毒71型对神经系统和免疫功能的影响及疫苗研究进展[J].实用儿科临床杂志,2012,27(22):1701-1704.
[5]Yao X,Mao QY,Liang ZL.Progress in research on enterovirus 71vaccine[J].Zhongguo Shengwu Zhipinxue Zazhi,2011,24(2):233-236.(in Chinese)姚昕,毛群颖,梁争论.肠道病毒71型疫苗的研发现状及进展[J].中国生物制品学杂志,2011,24(2):233-236.
[6]Jiang MB,Lu J,Mou W,et al.Disease burden of hand,foot-and-mouth disease and research progress of EV-A71vaccine[J].Shanghai Yufang Yixue,2016,28(6):382-384.(in Chinese)姜铭波,陆瑾,牟文,等.手足口病疾病负担和EV71疫苗的研究进展[J].上海预防医学,2016,28(6):382-384.
[7]Li RC,Liu LD,Zhao JM,et al.An inactivated enterovirus71vaccine in healthy children[J].New Engl J Med,2014,370(9):829-837.
[8]Liu LD,Zhao M,Liang ZL,et al.Immunity and clinical efficacy of an inactivated enterovirus 71vaccine in healthy Chinese children:a report of further observations[J].BMC Med,2015,13(1):226.
[9]Sun S,Gao F,Mao Q,et al.Immunogenicity and protective efficacy of an EV-A71virus-like particle vaccine against lethal challenge in newborn mice[J].Hum Vacc Immunother,2015,11(10):2406-2413.
[10]Zhang Y,Liu LD,Dong CH,et al.Consistency of various batches of inactivated EV-A71vaccine[J].Zhongguo Shengwu Zhipinxue Zazhi,2015,28(3):223-227.(in Chinese)张莹,刘龙丁,董承红,等.不同批次肠道病毒71型(EV71)灭活疫苗的一致性分析[J].中国生物制品学杂志,2015,28(3):223-227.
[11]Liu ZL,Li H,Long RX,et al.Research of CoxAl6antigen detection and preliminary application[J].Zhongguo Weisheng Jianyan Zazhi,2013,23(4):801-805.(in Chinese)刘正玲,李华,龙润乡,等.CoxA16抗原检测试剂研究及初步应用[J].中国卫生检验杂志,2013,23(4):801-805.
[12]Reed Z,Cardosa MJ.Status of research and development of vaccines for enterovirus 71[J].Vaccine,2016,34(26):2967-2970.
[13]Lin SY,Chung YC,Chiu HY,et al.Evaluation of the stability of enterovirus 71virus-like particle[J].J Biosci Bioeng,2014,117(3):366-371.