不同因素HCV结构区基因重组体诱发小鼠产生抗体的影响

The Effect of Different Administrations of pCD-HCV 1 on Antibody Responses in Mice Immunized by Gene Recombinant of HCV Structure Region

寻求ＨＣＶ基因免疫的最佳动物实验方法，探讨不同处理因素对基因重组体ｐＣＤ－ＨＣＶ１诱发小鼠产生抗体的影响。方法：用分子生物学技术构建丙型肝炎病毒基因重组体ｐＣＤ－ＨＣＶ１，采用不同免疫次数、途径、剂量和不同处理方法免疫Ｂａｌｂ／ｃ小鼠。结果：重组体ｐＣＤ－ＨＣＶ１经肌肉１、２、３和４次免疫小鼠后（１００μｇ／只，ｎ＝１２），抗体水平分别为０１８３±０００６、０４２８±００５、０７０７±００８和０７７３±００７（Ａ值，下同）。其中４次免疫小鼠的抗体水平最高；ｐＣＤ－ＨＣＶ经灌胃、腹腔注射、皮下注射和肌肉注射（１００μｇ）不同途径分别免疫小鼠（ｎ＝６）抗体水平依次为０１３８±０００５、０１７８±００７、０２３３±００８和０６９１±００５，ＳＮＫ统计显示，肌肉注射途径与其他３种注射途径相比，抗体水平有显著差异（Ｐ＜０００１）；以不同剂量（１０、５０和１００μｇ）分别免疫小鼠（ｎ＝８），抗体水平依次为０１１±００９、０３３±００４和０７０±００７；各组间差异有显著性意义（Ｐ＜００１）；用普鲁卡因１００μｇ（０．４ｍｇ）肌肉注射小鼠２４ｈ后，再肌肉、皮下注射ｐＣＤ－ＨＣ?

Objective: To seek the optimal experiment methods of animal with HCV gene immunity and explore the effect of different administrations of pCD-HCV 1 recombinant on the antibody responses in mice. Methods:Recombinant pCD-HCV 1 constructed by gene engineering was injected into muscles of BALB/c mice with different times, routes, doses and treament methods.Results:After recombinant pCD-HCV 1 (100 μg/mouse) were inoculated into mice once, twice, three times and four times, the serum antibody level reached to 0.183±0.06,0.428±0.05, 0.707±0.08,0.773±0.07 and 0.183±0.06, respectively. The antibody level in mice ( n =12) was the highest is all groups after inoculating four times; pCD-HCV 1 was perfused into stomach or inoculated into mice by i.p., s.c.and im (100 μg,three times) in different routes ( n =6) and the antibody levels were 0.138±0.05,0.78±0.07,0.233±0.08,0.690±0.05,respectively. After mice were inoculated with different doses (10 μg,50 μg and 100 μg) of pCD-HVC 1 the antibody levels were 0.110±0.09, 0.330±0.04 and 0.700±0.07, respectively, and there was a significant difference ( P <0.01). After mice were injected with procaine (100 μg,0.4 mg) and pCD-HCV 1, antibody levels were higher than those immunizated directly with recombinant pCD-HVC 1 with same dose.Conclusion: The results may provide a reference data deserved for seeking the optimal immune method for developing HCV DNA vaccine in mice modle.