白细胞介素-2、γ干扰素净化模拟白血病缓解期骨髓及用PCR检测净化效果

The purging effect of interleukin 2 and interferon γ on imitative bone marrow in remission and the detection by PCR of the effect

目的：探讨白细胞介素２（ＩＬ－２）、γ－干扰素（ＩＦＮ－γ）用于自体骨髓中残留白血病细胞的净化及用聚合酶链反应（ＲＴ－ＰＣＲ）方法检测净化效果。方法：将ＩＬ－２、ＩＦＮ－γ与模拟慢性粒细胞白血病（ＣＭＬ）缓解期的骨髓在培养体系中分别培养１～６ｄ，通过ＲＴ－ＰＣＲ方法来检测缓解期骨髓中ｂｃｒ／ａｂｌｍＲＮＡ消失情况。结果：模拟缓解期ＣＭＬ骨髓经与ＩＬ－２、ＩＦＮ－γ２４ｈ、４８ｈ和７２ｈ培养后用ＲＴ－ＰＣＲ方法仍可检测到ｂｃｒ／ａｂｌｍＲＮＡ，而经６ｄ培养后则已测不到。若将ＩＬ－２、ＩＦＮ－γ合用，经２４和４８ｈ培养的骨髓可检测到ｂｃｒ／ａｂｌｍＲＮＡ，但经７２ｈ和６ｄ培养后便检测不到，而对照组在１～６ｄ培养后仍可检测到该基因。结论：初步实验结果表明ＩＬ－２、ＩＦＮ－γ对缓解期骨髓中的Ｋ５６２白血病细胞系具有净化作用，若两者合用具有协同作用，ＲＴ－ＰＣＲ可作为评估净化效果较为敏感和较为可靠的检测方法。

Objective:To Evaluate the purging effect of interleukin 2 (IL 2) and interferon γ (IFN γ) on minimal residual leukemic cells during autologous bone marrow transplantation and the use of RT PCR technique in determining the purging effect. Methods:The imitative chronic myeloid leukemic bone marrow in remission had been cultured in liquid mediam with IL 2 and IFN γ for 1-6 days, and the puring effect was determined by detection of the bcr/ab1 mRNA. Results: The bcr/ab1 mRNA could be detected by RT PCR among bone marrow treated with IL 2 or IFN γ alone for 24, 48 and 72 hours, but it was negative after 6 days of culture. And among bone marrows treated with both IL 2 and IFN γ, the bcr/ab1 mRNA was positive when treated for 24 and 48 hours and negative when treated for 72 hours and 6 days. In those contrast bone marrows ,which neither IL 2 nor IFN γ was used, the bcr/ab1 mRNA could still be detected after 6 days of culture. Conclusion:IL 2 and IFN γ can be used to purge K 562 leukemic cells in bone marrow in remission and they have coordinate effect when used together. RT PCR is a sensitive and reliable method to evaluate the purging efftec.