以α甘露聚糖肽为佐剂的EV71全病毒灭活抗原黏膜免疫小鼠的抗体应答

基础医学与临床 ›› 2013, Vol. 33 ›› Issue (11): 1435-1439.

以α甘露聚糖肽为佐剂的EV71全病毒灭活抗原黏膜免疫小鼠的抗体应答

荆晶¹,孙强²,冯素玲³,赵兴华³,廖怡然³,王冰³,黎卫平⁴

1. 兰州大学基础医学院
2. 西安交通大学药理系
3. 西安交通大学医学院
4. 兰州大学基础医学院病理研究所

收稿日期:2013-01-15 修回日期:2013-03-27 出版日期:2013-11-05 发布日期:2013-10-28
通讯作者: 黎卫平 E-mail:liwp@lzu.edu.cn
基金资助:
国家高技术研究发展计划资助项目（863计划）

Immunization response of enterovirus 71 whole virus inactivation antigen combined with PA adjuvant by nasal mucosa in mice

Received:2013-01-15 Revised:2013-03-27 Online:2013-11-05 Published:2013-10-28
Supported by:
The National High Technology Research and Development Program of China (863 Program ）

摘要/Abstract

摘要： 目的肠道71型全病毒灭活抗原(EV71 Ag)联合α-甘露聚糖肽（PA）佐剂滴鼻免疫，观察全身免疫应答及持续时间。方法 5~6周龄BALB/c雌性小鼠随机分为阴性对照组，阳性对照组，高剂量疫苗组，低剂量疫苗佐剂组和高剂量疫苗佐剂组，每组10只。阴性对照组以PBS滴鼻，阳性对照组以EV71Ag肌注给药，免疫组以EV71 Ag单独和不同剂量分别配伍PA滴鼻给药。首次给药后第0、2、3、4、5和6周采集血清及鼻肺灌洗液，ELISA法检测血清特异性IgG抗体水平及鼻肺灌洗液sIgA抗体水平，MTT法检测淋巴细胞增殖，中和抗体实验检测小鼠血清中和抗体水平。结果各滴鼻组小鼠血清中均产生一定水平的特异性IgG抗体，高剂量疫苗佐剂组第三次免疫后其血清IgG（2.477±0.500）可达到阳性对照组（3.040±0.120）的80%，佐剂组体外淋巴细胞增殖率较阳性对照组有显著增高(P<0.05)，各滴鼻组血清中和抗体滴度均>8。结论 EV71全病毒灭活联合α-甘露聚糖肽佐剂免疫小鼠可诱导全身免疫应答，且α-甘露聚糖肽佐剂可增强其作用。

关键词: 粘膜免疫, EV71, 中和抗体, 特异性IgG抗体, sIgA抗体, 淋巴细胞增殖率

Abstract: Objective To study the systemic immune response intranasally immunized by type 71 whole virus inactivation antigen (EV71 Ag) combined with alpha mannan peptide (PA) adjuvant.. Methods Forty BALB/c mice that were five to six week old were randomly divided into five groups,10 mice per group. Immune groups were accepted EV71 Ag with or without adjuvants. Blood serum and bronchoalveolar lavages were collected at 0,2,3,4,5and6weeks since the first immune treatment.The level of IgG,IgA and neutralizing antibody in serum were detected by enzyme-linked immunosorbent assay(ELISA)and Neutralizing antibody tests repectively.MTT assay was involved to assess the proliferation of lymphocyte. Results The specific IgG antibody in serum could be detected in every group.There was no statistically significant different between the high-dose adjuvanted group and EV71Ag intramuscular injecting group（2.477±0.500 verse 3.040±0.120).EV71 Ag combined with alpha mannan peptide(PA)adjuvant caused lymphocyte proliferation in vitro,neutralizing antibody titer increased in all group.There were no significant difference between high-dose adjuvanted group and the EV71 Ag intramuscular injuecting group.Conclusion EV71 Ag combined with alpha mannan peptide(PA) adjuvant intranasally induced an effective systemic immune reponse.

Key words: Key words：Mucosal immunity, Enterovirus 71 antigen, Neutralizing antibody, specific IgG antibody, sIgA antibody, lymphocyte proliferation

中图分类号:

荆晶孙强冯素玲赵兴华廖怡然王冰黎卫平. 以α甘露聚糖肽为佐剂的EV71全病毒灭活抗原黏膜免疫小鼠的抗体应答[J]. 基础医学与临床, 2013, 33(11): 1435-1439.

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