不同来源T细胞及红细胞在胸腺肽类免疫增强剂活性检验方法中的替代性

Substitutability of T cells and erythrocytes from different sources in determination method of biological activity of thymosin immunopotentiator

目的研究不同来源的T细胞及红细胞在胸腺肽类免疫增强剂活性检验方法中的替代性。方法T细胞活性测定法-脱E受体法(E-玫瑰花结试验)参照国家食品药品监督管理局国家药品标准WS1-(X-001)-2002Z进行,采用豚鼠胸腺T细胞作为T细胞来源,兔红细胞作为红细胞来源,并对豚鼠周龄、脱E条件、兔红细胞保存方式进行优化;采用Jurkat细胞作为T细胞来源,绵羊红细胞作为红细胞来源,并对脱E时间、给药浓度、给药时间进行优化。结果豚鼠胸腺T细胞与兔红细胞可形成玫瑰花结,最佳试验条件为:选取4~7周龄豚鼠胸腺T细胞,45℃孵育1 h脱去E受体,用阿氏液保存兔红细胞;Jurkat细胞与绵羊红细胞可形成玫瑰花结,最佳试验条件为:45℃孵育30 min脱去E受体,1 mg/mL给药。结论豚鼠胸腺T细胞-兔红细胞及Jurkat细胞-绵羊红细胞的组合可形成玫瑰花结,但对样品的反应性不如猪胸腺T细胞-绵羊红细胞的组合。

Objective To investigate the substitutability of T cells and erythrocytes from different sources in the method for determination of biological activity of thymosin immunopotentiator.Methods T cell activity assay-E receptor detach method(E-rosette assay)was carried out according to national drug standard WS1-(X-001)-2002 Z approved by the State Food and Drug Administration.Guinea pig thymus was used as the source of T cells while rabbit erythrocytes as the source of erythrocytes.Conditions such as the age of guinea pig,the removal of E-receptor and the storage of rabbit erythrocytes were optimized.Meanwhile,Jurkat cells were used as the source of T cells while sheep erythrocytes as the source of erythrocytes,and the time for the removal of E-receptor as well as the concentration and incubation time of thymosin were optimized.Results T cells from guinea pig thymus formed rosettes with rabbit erythrocytes,of which the experimental condition was optimized as follows:the T cells from thymus of guinea pigs at ages of 4~7 weeks were incubated at 45℃for 1 h to remove the E-receptor and incubated with rabbit erythrocytes preserved in Alsever solution.However,Jurkat cells formed rosettes with sheep erythrocytes,of which the experimental condition was incubation at45℃for 30 min to remove E-receptor then incubation with thymosin at a concentration of 1 mg/mL.Conclusion Either the combination of T cells from guinea pig thymus-rabbit erythrocytes and Jurkat cells-sheep erythrocytes could form rosettes,of which the reactivities to thymosin were lower than that of combination of T cells from porcine thymussheep erythrocytes.