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目的探讨壳寡糖(chitooligosaccharide,COS)佐剂对甲型肝炎病毒(hapatitis A virus,HAV)疫苗诱导小鼠体液免疫应答的影响。方法将ICR小鼠随机分成9组:甲型肝炎减毒活疫苗Hep A-l 18 EU+不同剂量COS(100μg、500μg、1 mg、2.5 mg、5 mg、10 mg)组、阴性对照组(生理盐水)、抗原对照组(Hep A-l 18 EU)和铝佐剂对照组[Hep A-l18 EU+Al(OH)3200μg],每组6只,各组均经皮下注射ICR小鼠,200μl/只。于免疫后的第4、8、12和16周,采用间接ELISA法检测小鼠血清中抗-HAV特异性Ig G抗体水平。在实验期间,观察小鼠的健康状况,并于免疫16周后,取COS最佳剂量组及阴性对照组小鼠肾脏、脾脏、肝脏、肺脏、心脏组织,制备病理切片,进行对比观察。结果除阴性对照组外,各组小鼠免疫4周后均产生抗-HAV Ig G抗体,并随着时间的延长呈上升趋势,在第8周时均达峰值,随后逐渐下降;COS 2.5 mg、5 mg和10 mg组小鼠血清在各时期的抗体水平均显著高于抗原对照组和铝佐剂对照组(P<0.05),且能维持较长时间,至16周抗体水平仍与铝佐剂对照组相当,差异无统计学意义(P>0.05),其中COS5 mg组在整个试验过程中产生的抗体水平均最高,且与2.5 mg组峰值水平差异有统计学意义(P<0.05),与10 mg组水平相当,至免疫后16周,COS 5 mg组与10 mg组抗体水平差异有统计学意义(P<0.05)。COS最佳剂量为5 mg/只。试验期内,观察小鼠各项生理指标未见异常,COS 5 mg组小鼠心脏、肝脏、脾脏、肺脏、肾脏组织均未观察到病变。结论 COS可显著增强HAV疫苗诱导小鼠的体液免疫应答,有望开发为一种可替代铝佐剂的新型佐剂。
Objective To investigate the effect of chitooligosaccharide (COS) adjuvant on humoral immune response induced by hapatitis A virus (HAV) vaccine in mice. Methods The ICR mice were randomly divided into 9 groups: Hep Al 18 EU + live attenuated Hepatitis A vaccine group (100 μg, 500 μg, 1 mg, 2.5 mg, 5 mg, 10 mg), negative control group , Hep Al 18 EU and aluminum adjuvant control group [Hep A -18 EU + Al (OH) 3200 μg] with 6 mice in each group. All mice were injected subcutaneously with ICR mice at 200μl / mouse. At 4, 8, 12 and 16 weeks after immunization, the level of anti-HAV specific Ig G antibody in serum of mice was detected by indirect ELISA. During the experiment, the mice were observed for their health status. After 16 weeks of immunization, the kidney, spleen, liver, lung and heart tissues of mice in COS optimal dose group and the negative control group were taken for pathological observation. Results In addition to the negative control group, anti-HAV Ig G antibody was raised in all groups after 4 weeks of immunization and increased with time, reaching the peak value at the 8th week and then gradually decreasing. COS 2.5 mg, 5 mg and Antibody levels of 10 mg group serum at each time were significantly higher than that of the antigen control group and the aluminum adjuvant control group (P <0.05), and can maintain a long time, the antibody level is still 16 weeks and aluminum adjuvant control group (P> 0.05). COS5 mg group had the highest antibody level during the whole experiment, and the difference was statistically significant (P <0.05) with the peak value of 2.5 mg group The level of antibody in COS 5 mg group was significantly higher than that in 10 mg group at 16 weeks after immunization (P <0.05). The optimal dose of COS is 5 mg / bird. During the experimental period, no abnormalities were observed in all the physiological indexes of the mice. No lesions were observed in the heart, liver, spleen, lung and kidney of the COS 5 mg group. Conclusion COS can significantly enhance the humoral immune response induced by HAV vaccine and is expected to be a novel adjuvant that can replace aluminum adjuvant.