摘要
目的 初步探讨粘附NK细胞的制备方法及免疫学特性。方法 首先经淋巴细胞分离液、贴壁粘附及尼龙毛柱粘附对外周血NK细胞进行初步分离 ,然后联合使用Percoll不连续密度梯度离心法和T细胞Panning法进一步纯化NK细胞 ,应用流式细胞仪鉴定NK细胞并检测其纯度。rhIL 2 (6 0 0 0IU ml)诱导纯化的NK细胞 (R NK)转变为粘附NK(A NK)细胞 ,然后利用细胞计数法、MTT法和RT PCR方法检测A NK细胞的增殖和杀伤等免疫学特性。结果 经过纯化、诱导获得较高纯度、较高活性的A NK细胞。结论 初步建立了利用人外周血NK细胞制备A
Objective To investigate the method of fast harvest of IL-2-induced adherent human natural killer cells (A-NKs). Methods Peripheral blood mononuclear cells(PBMCs)were separated by centrifugation on Ficoll-Hypaque gradients. NK cells were then purified by Percoll density gradient centrifugation and T cell panning after passing through a nylon wool column. The purified NK cells were incubated in the presence of IL-2 (6 000IU/ml)and after one hour of activation, A-NKs were isolated from nonadherent natural killer cells(NA-NKs). Then the adherent kinetics, proliferation and cytotoxicity of A-NKs were analyzed by cell counting and MTT. RT-PCR was used to determine the mRNA of two cytotoxic molecules from A-NKs. Results A-NKs generated in this culture were always above 95% TCRαβ - CD56 +,as determined by flow cytometry. The proliferation and cytotoxicity of A-NKs were much higher than those of R-NKs and NA-NKs. Conclusion A method of harvesting A-NKs with high purity and high cytotoxicity was set up.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2002年第2期225-228,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目 ( 39870 72 9)
( 39970 82 8)
( 39970 6 94)
