传代人淋巴细胞免疫猪的免疫效果初探  

Evaluation of immune effect on continuous human T-lymphocyte cell line as antigen to produce anti-human T lymphocyte porcine plasma

作　　者：余健 熊小培 杨帆 吴笛 YU Jian;Xiong Xiao-pei;YANG Fan;WU Di(Research and Development Department,Yujin Bio-Pharma Wuhan CNBG Co.,Ltd.,Wuhan 430207,Hubei Province,China)

机构地区：[1]武汉中生毓晋生物医药有限责任公司科研开发部

出　　处：《微生物学免疫学进展》2019年第6期47-51,共5页Progress In Microbiology and Immunology

摘　　要：目的用传代人淋巴细胞替代人T淋巴细胞进行猪的免疫,用于试制抗人T细胞猪免疫球蛋白(anti-human T lymphocyte porcine immunoglobulin,P-ATG)免疫猪血浆,并评价其免疫效果。方法大量培养传代人淋巴细胞至免疫所需浓度及数量,采用常规猪免疫程序进行2次基础免疫及1次加强免疫,获得免疫猪血浆,用E玫瑰花环形成抑制试验和淋巴细胞毒试验进行效价检测。结果3批免疫猪血浆的E玫瑰花环形成抑制试验结果均达到1∶1000(花环形成率均小于对照组平均花环形成率的75%),淋巴细胞毒试验结果均达到1∶500(淋巴细胞死亡率均大于20%),效价均达到《中华人民共和国药典》2015版(三部)中生产用免疫猪血浆的效价标准。结论传代人淋巴细胞可作为人T淋巴细胞的替代免疫原进行猪的免疫,获得了效价合格的免疫猪血浆,用于P-ATG的制备。Objective To prepare the porcine plasma anti-human T lymphocyte immunoglobulin(P-ATG)with continuous human T-lymphocyte cell line as an antigen to replace primary human T lymphocytes,and to evaluate its immune effect.Methods To the distred concentration and quantity from continuous human T-lymphocyte cell needed in immunization was produced on a large-scale cultivation.Porcines were immunized by the routine immunization procedure with two basic and one boost injections.The antibody titer of immune porcine plasma was detected by E-rosette formation test and lymphocyte toxicity test.Results The immunized porcine plasma titer were 1∶1000 in E-rosette Inhibition test and 1∶500 in lymphocytic toxicity test in three batches,respectively.Corresponding to the antibody titer data,the E-rosette formation rate of test groups was lower than 75%formation rate of control groups.And lymphocyte mortality rate was greater than 20%.These data were in conformity with the standard of P-ATG in Chinese Pharmacopoeia.Conclusion The continuous human T-lymphocyte cell line can be used as an alternative antigen for immunization porcine in production of P-ATG.

关 键 词：传代人淋巴细胞 抗人T细胞猪免疫球蛋白 细胞培养 效价 E玫瑰花环形成抑制试验 淋巴细胞毒试验 免疫猪血浆 

分 类 号：R392-33[医药卫生—免疫学]