BSA作为动物源性医疗器械细胞免疫检测阳性对照物的研究  

Study on BSA as a positive control for cellular immunity evaluation of animal-derived medical devices

作　　者：杜晓丹[1] 方玉[1] 奚廷斐[1] 王春仁[1] 

机构地区：[1]中国药品生物制品检定所医疗器械检验中心,北京100050

出　　处：《药物分析杂志》2010年第7期1348-1350,共3页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的：目前国内外尚无针对动物源性医疗器械的免疫原性评价的统一标准。本研究参照ISO10993—20《医疗器械免疫毒性实验原则和方法》以及ASTM相关标准,如ASTMF1905-98《材料致免疫毒性测试方法选择标准规范》等,通过淋巴细胞增殖试验对BSA作为细胞免疫检测阳性物的相关技术问题进行了初步的研究,包括免疫剂量、免疫方式、取材时间等。为动物源性医疗器械的免疫原性评价实现标准化积累数据。方法：①实验动物：雌性BALB/c小鼠,8周。②免疫方式及免疫剂量：皮下注射高、中、低3个剂量的BSA（1000,500,100μg·mL^-1）或BSA与佐剂等量混合物,免疫2-3次,每次间隔2周。③淋巴细胞增殖试验：于不同时间点无菌取脾,分离脾细胞,进行淋巴细胞增殖实验。结果：①以BSA单独免疫小鼠,各剂量组OD值与阴性对照组比较差异均无统计学意义。②以BSA混合佐剂免疫小鼠,中、高剂量组OD值明显高于阴性对照组（p〈0.05）,且呈一定的剂量相关。③2次免疫后2周、3次免疫后2周BSA混合佐剂组OD值均明显高于阴性对照组（p〈0.05）。结论：将BSA与弗氏佐剂等量混合,以剂量为50-100μg·只^-1的BSA注射至小鼠皮下,每次间隔2周,2次或3次免疫后2周取材行淋巴细胞增殖试验可获得明显的阳性结果。Objective：Up to now,there is no uniform standard about immunogenicity evaluation of Animal-derived Medical devices at home and abroad.In this paper,some technical issues related to BSA as a positive control of cell mediated immune response was studied according to ISO10993-20,2006 and ASTM F1905-98（2003）.Methods：Animal：female BALB/c mice,8 weeks.Immunization of mice：mice were immunized subcutaneously with different doses of BSA（1000,500,100μg·mL^-1） or BSA plus adjuvant twice or three with an interval of two weeks.Cell mediated immune response：mice were sacrificed and spleen lymphocyte were isolated,lymphocyte Proliferation test was carried on by MTT assay or Alamablue assay.Results：①There was no significant different of OD values between negative group and BSA groups after the mice immunized by BSA alone（p〉0.05）.②The OD values of middle dose and high dose group were significant higher than that of negative group after the mice immunized by BSA plus adjuvant（p〈0.05）,and showed some dose related.③The OD values of BSA groups were significant higher than that of negative group after the second or third immunization by BSA plus adjuvant for two weeks（p〈0.05）.Conclusion：The results indicated that significant cell mediated immune responses can be found if the mice were immunized twice for 2 weeks or three for 2 weeks by BSA plus adjuvant with the dose of 50-100μg/animal.

关 键 词：牛血清白蛋白 细胞免疫 阳性对照物 动物源性医疗器械 

分 类 号：R917[医药卫生—药物分析学]