4-苯基咪唑+氢氧化铝复合佐剂对甲型肝炎减毒活疫苗诱导小鼠体液免疫应答的影响

Impact of 4-Phenylimidazole Combined with Aluminum Hydroxide on Humoral Immune Response Induced by Hepatitis A Virus Attenuated Live Vaccine in Mice

目的研究4-苯基咪唑(4-PI)+氢氧化铝[Al(OH)_3]复合佐剂对甲型肝炎减毒活疫苗(HepA-1)诱导的小鼠体液免疫应答的影响。方法实验设置7个分组,分别是:阴性对照组(1×PBS),铝佐剂组[HepA-1+Al(OH)_3],疫苗组(HepA-1),M1[HepA-1+Al(OH)_3+4-PI500μg]组,M2[HepA-1+Al(OH)_3+4-PI 1 mg]组,M3[HepA-1+Al(OH)_3+4-PI 1.5 mg]组和M4(HepA-1+4-PI 1mg)组,200μL HepA-1,24μL Al(OH)_3,随机分配小鼠,7只/组,分别皮下注射300μL,接种一次。用间接酶联免疫吸附法测试抗-甲型肝炎病毒(HAV)IgG抗体滴度,检测时间为免疫后4周、8周、12周、16周。实验过程中,观察和记录小鼠的健康情况。结果除阴性对照组,其余各组4个时间点均检测到抗-HAV IgG抗体,12周达到峰值。M2组在整个检测阶段抗体水平最高,显著高于疫苗组(t=4.449、3.633、2.565、6.809,P<0.05)和M4组(t=6.256、4.796、4.153、4.113,P<0.05),且第4周高于铝佐组(t=-2.877,P<0.05);M4组在4个时间点检测到的抗体水平与疫苗组相当。4-PI最佳剂量组是1 mg/只。实验全程观察到小鼠每项生理指标均正常。结论 4-PI+氢氧化铝复合佐剂能增强HepA-1诱导的小鼠体液免疫应答,有望开发成HepA-1新型复合佐剂。

Objective To investigate the impact of 4-phenylimidazole（4-PI） combining with aluminum hydroxide as an immunological adjuvant on humoral immune response in mice immunized with live hepatitis A virus （HAV） attenuated live vaccine （HepA-1）. Methods Seven experiment groups are set up, with 7 ICR mice per group randomly. Use 1 × phosphate buffer saline as negative control group, aluminum adjuvant group （HepA-1 200 μL＋AI（OH）3 24μL）, antigen group （HepA-1）, M4 （HepA-1＋4-PI 1 mg） as control, in which mice were injected subcutaneously with 300 μL, immunizing one time. The mice of three groups （M1, M2, M3） were immunized with live hepatitis A vaccine HepA-1 mixed with aluminum hydroxide,4-PI at concentrations of 500 μg, 1 mg, 1.5 mg respectively. The anti-HAV specific IgG antibody levels were tested by indirect ELISA method in the serum of mice after the first 4th, 8th, 12th, 16th week of injection. The health condition of mice were observed and record during the whole study. Results HAV-specific IgG levels of all mice were detected at four setting time point except negative control group and reached the maximum at 12th week. The IgG titers of group M2 mice were the highest at all detected time point and showed significant difference with those of antigen group （t=4.449, 3.633, 2.565, 6.809; P〈0.05）, group M4 （t=6.256,4.796, 4.153, 4.113; P〈0.05） and aluminum group at 4th week （t=2.877, P〈0.05）. The IgGs induced by group M4 was comparable to the antigen group in each test phase. The optimal dosage of 4-PI was 1 mg for each mouse. The physiological indexes in all the groups of mice was normal during the course of experiment. Conclusions The humoral immune responses induced by HepA-1 in mice was heightened by 4-PI combined with aluminum hydroxide,which had potential for developing a novel and compound HepA-1 adjuvant.