比格犬注射重组腺病毒Ad-HGF后血清中抗腺病毒抗体及中和抗体的检测

Detection of anti-adenovirus antibody and neutralization antibody in serum from Beagle dog after injected with recombinant adenovirus Ad-HGF

目的:观察Beagle犬重复给予Ad-HGF后血清中抗腺病毒抗体及抗肝细胞生长因子(hepatocyte growth factor,HGF)抗体的出现时间及消失规律.方法:将24只比格犬随机分为四组:正常对照组、手术对照组、Ad-HGF小剂量组(2.4×107pfu/kg)和Ad-HGF大剂量组(2.4×108pfu/kg).Ad-HGF各组犬给予不同剂量Ad-HGF共14次.在给药2次后分别利用病毒中和反应和ELISA方法检测血清中抗腺病毒抗体和抗HGF抗体.结果:手术组、正常对照组及实验组动物于给药前及给药3次时血清细胞均未出现明显中和反应.给药4次后各剂量组动物血清均对腺病毒有不同程度中和作用,至停药后12wk抗体水平很低或已不存在.各时间点血清样品中未检测到抗HGF抗体.结论:犬肌注Ad-HGF4次后动物血清中检测到了抗腺病毒抗体,停药4wk后抗体水平有不同程度下降,直至12wk后血清中腺病毒抗体基本消失.

AIM： To observe the emergence time and the disappearance rules of anti-adenovirus antibody and anti-hepatocyte growth factor（HGF） antibody in serum from Beagle dogs injected repeatedly with recombinant adenovirus Ad-HGF. METHODS： Twenty-four Beagle dogs were randomly divided into 4 groups： normal control group, operated control group, Ad-HGF low dosage[ （2.4 × 10^7） pfu/kg] group, Ad-HGF high dosage [ （2.4 × 10^8 ） pfu/kg] group. Ad-HGF at different doses was injected locally into muscles of Beagle dogs in Ad-HGF groups for total 14 times. After injection twice, serum anti-adenovirus antibody was measured in 293 cells by viral neutralization test in dog blood taken at every week. At the same time points, the anti-HGF antibody was detected by ELISA method. RESULTS： No obvious neutralization reaction in 293 cells was observed in serum from operated, control animals and experimental animals before injection and after injection for 3 times. After injection for 4 times, the serum of all Ad-HGF animals could neutralize adenovirus at different degree. The level of anti-adenovirus antibody in serum fell markedly at the 4th week after Ad-HGF withdrawal, and fell basically to ＂ ± ＂ at the 12th week. NO anti-HGF antibody was detected at all time points. CONCLUSION： Anti-adenovirus antibody in serum can be detected after about 4-time injections of Ad- HGF. The level of anti-adenovirus antibody in serum fell at different degree at the 4th week after termination of Ad-HGF injection. At the 12th week after stopping the administration, the antibody nearly vanished.