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目的探讨尿苷二磷酸(Uridine diphosphate,UDP)与尿苷三磷酸(Uridine triphosphate,UTP)两种佐剂对HAV抗原和HBsAg诱导小鼠体液免疫应答的影响。方法分别在HBsAg(1μg)和HAV抗原(18 EU)中加入不同浓度的UDP和UTP(HBsAg组:UDP和UTP均500μg及1、2、5、10 mg,HAV抗原组:UDP和UTP均500μg及1、2 mg),均经腹部皮下多点注射免疫ICR小鼠(HBsAg组:2针,间隔2周,0.1 ml/只;HAV抗原组:单针免疫,0.2 ml/只),并设空白对照组(生理盐水100μl)、抗原组对照组(HBsAg 1μg;HAV抗原18 EU)、铝佐剂组对照组(铝佐剂300μg)、UDP+铝佐剂复合组(HBsAg 1μg+铝佐剂300μg+UDP 2 mg;HAV 18 EU+铝佐剂300μg+UDP 2 mg)和UTP+铝佐剂复合组(HBsAg 1μg+铝佐剂300μg+UTP 2 mg;HAV 18 EU+铝佐剂300μg+UTP 2 mg),分别于末次免疫后(HBsAg组:第4、8、12和16周;HAV抗原组:第4、8、12周)采血,分离血清,ELISA法检测小鼠血清中抗-HBsAg IgG和抗-HAV IgG水平。免疫18周后,分别取HBsAg组中UDP和UTP最佳浓度组及空白对照组小鼠心脏、肝脏、脾脏、肾脏及肺组织进行病理观察。结果除空白对照组小鼠血清检测不到抗-HBsAg IgG外,其余各组小鼠血清抗-HBsAg IgG及抗-HAV IgG水平均在第8周达到峰值,以后逐渐下降。末次免疫后,UDP及UTP各组抗-HBsAg IgG及抗-HAV IgG水平均高于抗原对照组,差异均有统计学意义(P均<0.05)。HBsAg组UDP+铝佐剂复合组产生的IgG水平明显高于抗原对照组、铝佐剂对照组及UDP和UTP各浓度组(P<0.05);HAV抗原组UTP+铝佐剂复合组产生的IgG水平明显高于抗原对照组、铝佐剂对照组及UDP和UTP各浓度组(P<0.05)。在两种抗原中,UDP和UTP的最佳浓度均为2 mg/只。在设置的浓度范围内,UDP和UTP佐剂均未观察到毒性反应。结论UDP和UTP均能明显增强HBsAg及HAV抗原诱导小鼠的体液免疫应答。
Objective To investigate the effects of Uridine diphosphate (UDP) and Uridine triphosphate (UTPT) on the humoral immune responses induced by HAV antigen and HBsAg in mice. Methods Different concentrations of UDP and UTP were added to HBsAg (1μg) and HAV antigen (18 EU) respectively (HBsAg group: 500μg and 1,2,5,10 mg for both UDP and UTP, HAV antigen group: 500μg for both UDP and UTP And 1, 2 mg) were injected intraperitoneally subcutaneously in mice ICR mice (HBsAg group: 2-pin interval of 2 weeks, 0.1 ml / only; HAV antigen group: single needle immunization, 0.2 ml / only) The control group (HBsAg 1μg; HAV antigen 18 EU), aluminum adjuvant group (aluminum adjuvant 300μg), UDP + aluminum adjuvant group (HBsAg 1μg + aluminum adjuvant 300μg + UDP 2 mg; HAV 18 EU + aluminum adjuvant 300 μg + UDP 2 mg) and UTP + aluminum adjuvant combination group (HBsAg 1 μg + aluminum adjuvant 300 μg + UTP 2 mg; HAV 18 EU + aluminum adjuvant 300 μg + UTP 2 mg) After the last immunization (HBsAg group: 4th, 8th, 12th and 16th weeks; HAV antigen group: 4th, 8th and 12th weeks), blood serum was collected and the serum levels of anti-HBsAg IgG and anti-HAV IgG Level. After 18 weeks of immunization, the pathological changes of heart, liver, spleen, kidney and lung were observed in the optimal concentrations of UDP and UTP in HBsAg group and the blank control group respectively. Results The serum levels of anti-HBsAg IgG and anti-HAV IgG in the other groups all reached the peak value at the 8th week except the anti-HBsAg IgG in the serum of the blank control group, and then decreased gradually. After the last immunization, the levels of anti-HBsAg IgG and anti-HAV IgG in both UDP and UTP groups were significantly higher than those in the antigen control group (all P <0.05). The level of IgG produced by HBsAg + UDP + aluminum adjuvant group was significantly higher than that of antigen control group, aluminum adjuvant control group and UDP and UTP groups (P <0.05). The levels of IgG produced by HAV antigen group combined with UTP + aluminum adjuvant Significantly higher than the antigen control group, the aluminum adjuvant control group and the concentration of UDP and UTP groups (P <0.05). The best concentration of UDP and UTP for both antigens was 2 mg / mouse. No toxic reaction was observed in both UDP and UTP adjuvants within the concentration range set. Conclusion Both UDP and UTP can significantly enhance the humoral immune response induced by HBsAg and HAV antigens in mice.