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目的探讨CpG寡聚脱氧核苷酸(CpG oligodeoxynucleotides,CpG ODN)对狂犬病疫苗(rabies vaccine,RV)免疫效果的影响。方法以市售狂犬病疫苗为抗原,分别以低、中、高剂量的CpG ODN(1.25、5、20μg/剂)及其与氢氧化铝[(Al(OH)_3]联合的复合物作为疫苗佐剂,按Essen程序经腿部肌肉免疫BALB/c小鼠。免疫前2 d及首次免疫后分别于第6、8、10、14、35和42天采血,分离血清,通过快速荧光灶抑制试验(rapid fluorescent focus inhibition test,RFFIT)检测血清中抗狂犬病病毒中和抗体效价;并经ELISPOT法分析首次免疫第6和14天小鼠脾细胞分泌IFNγ的情况。结果低、中剂量的CpG ODN及其与Al(OH)_3联合使用均可使小鼠在首次免疫后第8天产生具有保护水平的中和抗体,第10天阳转率均达100%;在首次免疫后第35及42天,小鼠产生抗狂犬病病毒中和抗体水平均明显升高(P<0.05);在首次免疫后第6及14天,低、中剂量的CpG ODN可明显提高小鼠脾细胞的IFNγ分泌水平(P<0.05)。结论 CpG ODN对RV具有免疫增效作用,可能成为一种新型的RV佐剂。
Objective To investigate the effect of CpG oligodeoxynucleotides (CpG ODN) on rabies vaccine (RV) immunization. Methods Commercially available rabies vaccine was used as antigen and CpG ODN (1.25, 5 and 20μg / dose) and its complex with aluminum hydroxide [(Al (OH) 3] The BALB / c mice were immunized by the Essen procedure with muscle of the leg, and the blood was collected on the 6th, 8th, 10th, 14th, 35th and 42nd days before immunization and after the first immunization respectively, The serum anti-rabies neutralizing antibody titers were detected by rapid fluorescent focus inhibition test (RFFIT), and IFNγ secreted by splenocytes of mice on the 6th and 14th day after the first immunization was analyzed by ELISPOT.Results Low and medium doses of CpG ODN In combination with Al (OH) 3, mice with protective levels of neutralizing antibody were produced on the 8th day after the first immunization. The rates of positive conversion reached 100% on the 10th day. After the first immunization, 35 and 42 (P <0.05). At the 6th and 14th days after the first immunization, low and medium doses of CpG ODN could significantly increase the level of IFNγ secretion in mouse spleen cells (P <0.05) .Conclusion CpG ODN has an immunopotentiating effect on RV and may become a new RV adjuvant.