聚乙二醇化重组人粒细胞集落刺激因子大鼠和Beagle犬的免疫原性

目的：在进行聚乙二醇化重组人粒细胞集落刺激因子（PEG30-rhG-CSF）大鼠和Beagle犬重复给药毒性试验时,观察给药后动物血清中抗PEG30-rhG-CSF抗体的产生和抗体的中和活性,为非临床安全性评价的确切性以及临床给药周期提供依据。方法：大鼠分为赋形剂对照组、PEG30-rhG-CSF0.75、3.0、12mg/kg组,sc给药,隔日1次,连续2周;Beagle犬分为赋形剂对照组、PEG30-rhG-CSF0.25、1.0和4.0mg/kg,sc,每周1次,连续4周。采用ELISA法检测动物血清中抗PEG30-rhG-CSF的抗体,采用NFS-60细胞/MTT比色法检测抗体的中和活性。结果：大鼠在给药期和恢复期各剂量组动物血清中均未检测到抗PEG30-rhG-CSF的结合抗体。Beagle犬在给药的第1周和第2周,未检测到结合抗体,但在第4周时,低、中和高3个剂量组各有5只（5/6）的动物血清中检测到结合抗体,而抗体滴度与剂量无明显相关性。恢复4周后,仅高剂量组1只（1/2）动物出现抗体,且抗体滴度呈下降趋势。另外,在给药期和恢复期,血清中所产生的抗体均无中和PEG30-rhG-CSF的活性。结论：大鼠重复给予PEG30-rhG-CSF的毒性试验中,所有动物血清中均未检测到结合抗体和中和抗体;Beagle犬用药组的绝大部分动物血清中出现抗PEG30-rhG-CSF的抗体,但所产生抗体无中和活性。

Immunogenicity of PEGylated recombinant human granulocyte colony stimulating factor in rats and Beagle dogs

Objective： To investigate the anti-pegylated recombinant human granulocyte colony stimulating factor （PEG30-rhG-CSF） antibody formation and neutralizing antibody activity of PEG30-rhG-CSF in repeated-dose toxicity studies in rats and Beagle dogs. Methods： The rats received sc injection of PEG30-rhG-CSF at doses of 0 （placebo）, 0.75, 3.0, and 12 mg / kg （every other day dosing for up to 2 weeks） and Beagle dogs at doses of 0 （placebo）, 0.25, 1.0, and 4.0 mg / kg （weekly dosing for up to 4 weeks）, respectively. Anti-PEG30-rhG-CSF antibody in serum was measured using ELISA and the neutralizing antibody activity was determined by cell-based bioassay. Results： No anti-PEG30-rhG-CSF responses were observed in rats during the experiment. The antibody titers in serum samples were determined in dogs during the treatment and recovery period, but no neutralizing antibodies were found. There was no apparent dose-response in either incidence of antibody positivity or titer of antibody development in dogs, and titers sharply declined at the end of the recovery period. Conclusion： Antibodies to PEG30-rhG-CSF were not detected in the rats. The dogs have developed binding antibodies, but no neutralizing potency.