重组人粒细胞巨噬细胞刺激因子栓在大鼠和Beagle犬中的免疫原性

目的在大鼠和Beagle犬阴道给予重组人粒细胞巨噬细胞刺激因子(recombinant human granulocyte-macrophage colony-stimulating factor,rhGM-CSF)栓3个月的长期毒性试验中,检测给药后动物血清中是否产生抗rhGM-CSF抗体和抗体的中和活性。方法 SD雌性大鼠设rhGM-CSF栓0(赋形剂对照组)、0.24、0.96和3.84 mg/kg 4个组;Beagle雌性Beagle犬设0(赋形剂对照组)、0.07、0.28和1.12 mg/kg 4个组。每天给药1次,连续3个月,恢复期1个月。间接ELISA法检测动物血清中抗rhGM-CSF的结合抗体,TF-1细胞/MTT比色法检测抗体的中和活性。结果①大鼠从给药1个月起至恢复期结束,3个剂量组动物血清中均能检测到抗rhGM-CSF的抗体;低剂量和中剂量2组间动物血清的抗体滴度差异无统计学意义,但与高剂量组相比低剂量组和中剂量组差异有统计学意义。体外测活法显示,各剂量组在给药1个月后有部分动物产生中和抗体;给药3个月或恢复期1个月时全部受检动物均产生中和抗体。②Beagle犬的各剂量组动物血清中均未检测到抗rhGM-CSF的抗体。结论 rhGM-CSF栓经阴道局部连续给予3个月的长期毒性试验中,啮齿类动物大鼠血清中各剂量组均产生抗rhGM-CSF的抗体,且抗体可中和GM-CSF的生物活性;而非啮齿类动物Beagle犬血清中则未检测到抗rhGM-CSF抗体。rhGM-CSF栓对大鼠具有免疫原性。

Immunogenicity of recombinant human granulocyte-macrophage colony-stimulating factor suppository in rats and beagle dogs

Objective To investigate the anti-recombinant human granulocyte-macrophage colony-stimulating factor (anti-rhGM-CSF) antibody formation and neutralizing antibody activity of rhGM-CSF suppository in repeated-dose toxicity studies in rats and Beagle dogs. Methods The rats received vaginal suppository of rhGM-CSF at doses of 0 (placebo), 0.24, 0.96 and 3.84 mg/kg and Beagle dogs at a dose of 0 (placebo), 0.07, 0.28 and 1.12 mg/kg (daily dosing for up to 3 months), respectively. The titer of anti-rhGM-CSF antibody in serum was measured using indirect enzyme-linked immunosorbent assay (ELISA) and the neutralizing antibodies were determined by an in vitro bioassay. Results Antibodies to rhGM-CSF were detected in all the rats treated with rhGM-CSF suppository during the treatment and recovery period, and neutralizing antibodies were found in serum samples which developed binding antibodies. No anti-rhGM-CSF antibodies were observed in dogs during the experiment of all groups. Conclusion The rats serum contained rhGM-CSF binding antibodies which neutralized the biological activity of rhGM-CSF. Antibodies to rhGM-CSF were not detected in the Beagle dogs serum.