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目的研究烟酰胺腺嘌呤二核苷酸(Nicotinamide adenine dinucleotide,NAD)与氢氧化铝联合佐剂对HBsAg诱导小鼠体液免疫应答的影响。方法将NAD与氢氧化铝按不同剂量联合后,与不同剂量的HBsAg混合,分别经肌肉注射免疫ICR小鼠1针或2针(间隔2周),每只注射0.2 ml,并设阴性对照组(生理盐水)、单纯抗原对照组、单纯铝佐剂对照组和单纯NAD对照组,共23组,每组6只。分别于末次免疫后2、4、6、8、10、12周采血,分离血清,采用ELISA法检测小鼠血清中抗HBsAg IgG水平;试验期内,观察小鼠是否发生异常状况;14周后,取最佳剂量免疫组及阴性对照组小鼠心、肝、脾、肺组织做组织切片,进行病理分析。结果阴性对照组在各时间点均未检测到抗HBsAg IgG抗体,除联合佐剂12组外,其余各组血清抗HBsAg IgG抗体水平均在末次免疫后第6周达峰值,随着时间的延长,抗体水平呈下降趋势;联合佐剂14组(HBsAg 2μg+氢氧化铝100μg+NAD 2.5μg)产生的抗体水平最高,且持续时间较长,其对HBsAg的体液免疫增强效应显著优于单纯抗原对照组、单纯铝佐剂对照组及单纯NAD佐剂对照组(P<0.05)。试验期内,各组小鼠均未出现异常反应;最佳剂量免疫组(14组)及阴性对照组组织切片观察结果显示,均为发生病变。结论 NAD与氢氧化铝联合佐剂能显著增强HBsAg诱导的小鼠特异性体液免疫应答,在免疫剂量范围内,联合佐剂安全性良好。
Objective To investigate the effect of adjuvant combined with nicotinamide adenine dinucleotide (NAD) and aluminum hydroxide on humoral immune response induced by HBsAg in mice. Methods NAD and aluminum hydroxide were combined with different doses of HBsAg and mixed with different doses of HBsAg. ICR mice were immunized intramuscularly with 1 or 2 needles (interval 2 weeks) and injected with 0.2 ml each. Negative control group (Normal saline), simple antigen control group, simple aluminum adjuvant control group and simple NAD control group, a total of 23 groups, 6 in each group. Blood samples were collected at 2, 4, 6, 8, 10, and 12 weeks after the last immunization, respectively. Serum anti-HBsAg IgG levels were measured by ELISA. Abnormalities of mice were observed during the experimental period. , Take the best dose of immunization group and negative control group mice heart, liver, spleen, lung tissue sections, pathological analysis. Results The anti-HBsAg IgG antibody was not detected in the negative control group at all time points. The serum anti-HBsAg IgG levels in all the groups except the combination adjuvant group reached the peak at the 6th week after the last immunization. As time went by, , And the level of antibody showed a decreasing trend. The antibody produced by combination adjuvant 14 group (HBsAg 2 μg + aluminum hydroxide 100 μg + NAD 2.5 μg) had the highest level of antibody for longer duration, and its enhancement effect on humoral immunity of HBsAg was significantly better than that of the simple antigen control Group, simple aluminum adjuvant control group and simple NAD adjuvant control group (P <0.05). During the experimental period, no abnormal reaction occurred in all the mice in each group. The best dose immunization group (14 groups) and the negative control group tissue sections showed that all lesions occurred. Conclusion NAD and aluminum hydroxide adjuvant can significantly enhance the specific humoral immune response induced by HBsAg in mice, and the adjuvant is safe in the range of immunological dose.