摘要
目的研究人酸性成纤维细胞生长因子(Human acidic fibroblast growth factor,haFGF)和穿膜肽(Transcriptionalactivator protein,TAT)融合蛋白TAT-haFGF14-154在小鼠体内的急性毒性反应和免疫原性,初步评价其安全性。方法取昆明小鼠,单次经尾静脉注射10 mg/kg TAT-haFGF14-154,观察并记录14 d内小鼠的一般行为、中毒死亡情况和体重变化,各组织脏器中毒情况,HE染色观察各组小鼠大脑组织病理学变化。将900μg/kg TAT-haFGF14-154分别经静脉和鼻腔免疫昆明小鼠,每隔2 d给药1次,分别于给药后的第1、2、3、4周经眼眶采血,分离血清,ELISA法检测抗体效价;分别将高(300μg/kg)、中(100μg/kg)、低(35μg/kg)剂量的TAT-haFGF14-154经鼻腔免疫快速老化小鼠(SAM P8),每隔2 d给药1次,30 d后处死小鼠,经眼眶采血,分离血清,ELISA法检测抗体效价。结果在整个急性毒性试验过程中,小鼠无一例死亡,体重变化正常,大脑未见明显病理学改变;TAT-haFGF14-154对小鼠的最大耐受剂量大于10 mg/kg;昆明小鼠经鼻腔给予TAT-haFGF14-154后,第3周产生抗体,而静脉给药后第2周即有抗体产生;抗体产生的强度呈剂量依赖性。结论 TAT-haFGF14-154属于弱毒性药物,但具有免疫原性,静脉给药的免疫原性强于鼻腔给药。
Objective To investigate the acute toxicity and immunogenicity of fusion protein TAT-haFGFl14-154 consisting of human acidic fibroblast growth factor（haFGF） and transcriptional activator protein （TAT） in mice and preliminarily evaluate its safety. Methods Kunming mice were injected i.v. with a single dose of TAT-haFGF14-154 at 10 mg / kg, and observed for the general behavior, death caused by toxication, bodyweight and toxication in various organs within 14 d, and for pathological change in brain tissue by HE staining. Kunming mice were immunized with TAT-haFGF14-154 at 900 μg / kg by intravenous and intranasal mutes respectively, once 2 d for 30 d, of which the sera were collected at weeks 1, 2, 3 and 4 after immunization, and determined for antibody titer by ELISA. SAM P8 mice were immunized with TAT-haFGF1.154 at high （300 μg/kg）, moderate （100 μg/kg） and low （35μg / kg） dosages by intranasal route respectively, once 2 d, and killed 30 d later, of which the sera were separated and determined for antibody titer by ELISA. Results No deaths of mice were observed during acute toxicity test, while the change of bodyweight was within the normal range, and no obvious pathological change in brain tissue. The maximum tolerant dose of TAT- haFGF14-154 to mice was more than 10 mg/kg. Antibodies were induced in mice at week 3 after immunization with TAT-haFGF14-154by intranasal route and at week 2 after immunization by intravenous mute, of which the level was dose-dependent. Conclusion TAT- haFGF14-154 is a low toxic drug, of which the immunogenicity by intravenous route is higher than that by intranasal mute.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第10期1315-1318,共4页
Chinese Journal of Biologicals
基金
国家"重大新药创制"科技重大专项资助项目(2009ZX 09103-749)
广东省重大科技专项"重大新药创制"课题(2011A080502014)
关键词
急性毒性
免疫原性
人酸性成纤维细胞生长因子
穿膜肽
鼻腔给药
Acute toxicity
Immunogenicity
Human acidic fibroblast growth factor
Transmembrane peptide
Intranasal administration
