摘要
目的比较不同浓度配比的CpG ODN和Al(OH)3佐剂乙型肝炎疫苗的免疫原性。方法按乙肝表面抗原浓度为20μg/ml,分别加入3个浓度的Al(OH)3(300、400、500μg/ml)和CpG ODN(125、250、500μg/ml)佐剂,按不同配比制备9组不同佐剂浓度的疫苗样品,以不加CpG ODN佐剂的疫苗作为对照。将各组疫苗样品分别按1∶16、1∶32、1∶64、1∶128、1∶256共5个稀释度稀释后,经腹腔免疫BALB/c小鼠,第5周检测血清中抗-HBs水平,计算抗体阳转率及半数有效量(ED50);将各组疫苗样品分别于第0和28天经左前胫肌肉免疫BALB/c小鼠,分别于第1次免疫后第2、4、6、8、10周检测体液免疫应答水平,第5周检测细胞免疫应答水平。结果不同浓度CpG ODN和Al(OH)3的9组疫苗样品免疫小鼠后,ED50均小于0.2,对照组疫苗的ED50为0.235。不同浓度CpG ODN和Al(OH)3的9组疫苗样品小鼠血清抗体滴度均高于对照组,且抗体产生时间略提前;Al(OH)3浓度为400和500μg/ml时,疫苗诱导小鼠产生的抗体滴度与CpG ODN佐剂浓度呈正相关。对照组诱导产生的IFNγ水平均较其他疫苗组低;随着A1(OH)3浓度的升高,同一CpG ODN浓度的疫苗样品诱导IFNγ的水平逐渐下降;相同A1(OH)3浓度下,低浓度CpG ODN组斑点形成细胞数(spot-forming cell,SFC)均值较高,而高浓度组SFC均值较低。结论 A1(OH)3和CpG ODN作为双佐剂系统协同乙型肝炎疫苗,不仅能增强体液免疫应答,还能诱导IFNγ的表达,激活Th1细胞免疫应答,比传统A1(OH)3单佐剂乙型肝炎疫苗更具优势,其中CpG ODN和Al(OH)3浓度均为500μg/ml的疫苗制剂免疫原性最佳。
Objective To compare the immunogenicity of hepatitis B（HB）vaccines containing various concentrations of CpG ODN and aluminum hydroxide adjuvant. Methods Nine groups of HB vaccines were prepared by mixing 20 μg / ml HBsAg containing 300,400 and 500 μg / ml aluminum hydroxide with 125,250 and 500 μg / ml CpG ODN separately,using CpG ODN-free vaccine,containing 500 μg / ml aluminum hydroxide ＋ 20 μg / ml HBsAg,as control. The vaccine samples in various groups were diluted to dilutions of 1 ∶ 16,1 ∶ 32,1 ∶ 64,1 ∶ 128 and 1 ∶ 256,and immunized i. p. to BALB / c mice. The anti-HBs level in sera was determined at week 5 after immunization,based on which the antibody positive conversion rate and ED50were calculated. BALB / c mice were immunized i. m. with the vaccines in various groups on days 0 and 28,of which the humoral immune response levels were determined at weeks 2,4,6,8 and 10,while cellular immune response levels at week 5 after the first immunization. Results All the ED50values of vaccines in nine test groups were less than 0. 2,while that in control group was 0. 235. Compared with those in control group,all the serum antibody titers of mice immunized by the nine groups of vaccines were high,while the antibodies were produced slightly early. When the aluminum hydroxide concentrations were 400 and 500 μg / ml,the antibody titers induced by vaccines were positively related to the concentration of CpG ODN. The IFNγ level induced in control group was lower than those in test groups. Along with the increasing aluminum hydroxide concentration,the IFNγ induced by vaccines containing CpG ODN at the same concentrations decreased gradually. However,at the same aluminum hydroxide concentrations,the mean SFCs induced by vaccines containing CpG ODN at low concentrations were large,while those at high concentrations were small. Conclusion The combination of aluminum hydroxide and CpG ODN enhanced the humoral immune response induced by HB vaccine,induced the expression of IFNγ,and activated Th1 cellul
出处
《中国生物制品学杂志》
CAS
CSCD
2014年第5期596-600,共5页
Chinese Journal of Biologicals
基金
国家高技术研究发展计划-系列新型重组汉逊酵母乙型肝炎疫苗临床研究及产业化(2012AA02A404)
