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

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

目的肠道71型全病毒灭活抗原（EV71Ag）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EV71Ag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，MTr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全病毒灭活联合仪．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。

Objective To study the systemic immune response of animals immunized by type 71 virus antigen （EV71 Ag） combined with alpha mannan peptide （PA） adjuvant. Methods Forty BALB/c mice of five to six weeks old were randomly divided into five groups. Immune groups accepted EV71 Ag with or without adjuvants. Blood serum and bronchoalveolar lavages were collected at 0,2,3,4,5 and 6 weeks since the first immune treat- ment. The level of IgG, IgA and neutralizing antibody in serum was detected by enzyme-linked immunosorbent assay （ELISA） and Neutralizing antibody tests repectively. MTT assay was involved to assess the proliferation of lympho- cyte. Results The specific IgG antibody in serum was detected in all groups. There was no statistically significant different between the high-dose adjuvanted group and EV71 Ag intramuscular injecting group （2. 477±0. 500 vs 3.040± 0. 120）. EV71 Ag combined with alpha mannan peptide （PA）adjuvant caused lymphocyte proliferation invitro, neutralizing antibody titer increased in all groups. There were no significant difference between high-dose adju- vanted group and the EV71 Ag intramuscular injuecting group. Conclusions EV71 Ag combined with alpha man- nan peptide（PA） adjuvant intranasally induced an effective systemic immune reponse.