混合淋巴细胞反应的标准化

Standardisation of the mixed lymphocyte reaction.

作者信息

Yamamura M, Nikbin B, Hobbs J R

出版信息

J Immunol Methods. 1976;10(4):367-78. doi: 10.1016/0022-1759(76)90031-4.

Abstract

The mixed lymphocyte reaction has been optimised with Ficoll--Triosil separated lymphocytes: 100,000 responding cells with 100,000 mitomycin-C-blocked stimulating cells, in 0.2 ml of TC 199 with 10% AB-negative known normal serum in Cook microplates. After incubation at 37 degree C for 120 h, 2 muCi of [3H]thymidine was added, harvesting after a further 15 h at 37 degree C. A pool of stimulating cells, freshly separated from 5 normal subjects (usable up to 36 h if kept at 4 degree C), produced a Gaussian normal range of increment in CPM from 17,000--88,000, with a between-batch coefficient of variation for triplicates of 22%. For tests against stimulating cells from a single donor a transformation index above 1.64 was significant.

摘要

采用Ficoll - Triosil分离的淋巴细胞优化了混合淋巴细胞反应：在Cook微孔板中，将100,000个反应细胞与100,000个经丝裂霉素C处理的阻断刺激细胞混合，加入含10% AB阴性已知正常血清的0.2 ml TC 199培养基。在37℃孵育120小时后，加入2 μCi的[³H]胸腺嘧啶核苷，于37℃再孵育15小时后收获。从5名正常受试者新鲜分离的一组刺激细胞（若保存在4℃，最长可使用36小时），产生的CPM增加值呈高斯正态分布，范围为17,000 - 88,000，一式三份的批次间变异系数为22%。针对来自单一供体的刺激细胞进行检测时，转化指数高于1.64具有统计学意义。