Wistar大鼠肌内注射HPV疫苗重复给药毒性研究

摘要

目的:评价大鼠重复给药预防宫颈癌的HPV疫苗的安全性.方法:将大鼠按体重随机分为4组:阴性对照组、铝佐剂对照组,HPV疫苗低剂量组和高剂量组,分别在d1,d 22,d43,d 64肌内注射给药,恢复期4周,于给药前和给药后不同时间点进行各项毒理学指标检测.结果:各组动物的临床症状、体重、摄食量、体温、和尿检查各项指标均未见与给药相关的明显异常.在末次给药结束后,HPV给药组动物血液中性粒细胞及白细胞数量轻微升高,但在停药4周后恢复正常.抗核抗体检测结果显示各组动物各时间点血清均为阴性.疫苗所含3种抗原蛋白在大鼠体内均能诱导产生较高的抗体滴度,表明疫苗具有良好的免疫原性.组织病理学检查结果显示与给予供试品相关的病理学变化主要发生于注射部位、坐骨神经、肌肉(骨骼肌)以及腹股沟淋巴结,表现为不同程度的肉芽肿性炎症、间质纤维组织增生、水肿、炎性细胞浸润等,停药后逐渐减轻.结论:大鼠重复肌内注射HPV疫苗,具有良好的耐受性,无明显毒性反应剂量为人用剂量的2倍.本研究为下一步HPV疫苗进入临床试验奠定了基础.%Objective:To evaluate the preclinical safety of human papillomavirus vaccines (HPV) for preventing cervical cancer.Methods:Rats were randomly divided into four groups according to bodyweight,including control,aluminum adjuvant,low-and high-dose HPV vaccine groups.The animals were intramuscularly given vehicle or test articles on day 1,22,43 and 64 with 4-week recovery.The toxicological parameters were determined on different time points before and after administration.Results:Following repeated intramuscular administration of test articles,there were no obvious abnormalities in clinical symptom,weight,food consumption,temperature,and urinalysis in all groups.Mild increase of neutrophil and WBC counts were found in HPV vaccine-treated groups at the end of dosing period,but they returned to normal levels in the recovery period.No antinuclear antibody was detected in all groups.Three antigen component proteins induced high antibody responses in rats,suggesting these proteins had good immunogenicity.Histopathological examination showed that test article-related findings mainly concentrated at the injection site,sciatic nerve,muscle (skeletal muscles) and inguinal lymph nodes,which were characterized by different levels of granulomatous inflammation,interstitial fibrous tissue hyperplasia,edema,and inflammatory cells infiltration.All histopathological changes were gradually reduced over time.Conclusion:Repeated intramuscular dosing of HPV vaccines was well tolerated in rats,and no obvious toxicological responses were found.The dose without observed adverse effects was two times of human dose.These data will facilitate HPV vaccines to enter into clinical trials.