Simultaneous flow cytometric measurement of K-562 megakaryocytic differentiation and CD56+ large granular lymphocyte cytotoxicity
- 作者：Baker ; E.J. ; Ichiki ; A.T. ; Day ; N.E. ; Andrews ; R.B. ; Bamberger ; E.G. ; Lozzio ; C.B.
- 关键词：Human CD56 cells ; Human CD61 ; Phorbol ester ; K-562 ; Flow cytometry ; CML ; chronic myelogenous leukemia ; FBS ; fetal bovine serum ; LGL ; large granular lymphocytes ; MHC-I ; major histocompatibility complex class I ; mAb ; monoclonal antibody ; NK ; natural killer
- 刊名：Journal of Immunological Methods
- 出版年：2001
- 期刊代码：57_00221759
- 类别：imm
- 出版时间：July 1, 2001
- 卷：253
- 期：1-2
- 页码：37-44
- 文件大小：276 K
摘要
K-562 cells have the capacity to undergo multi-lineage differentiation, which may be crucial to their ability to serve as target reservoirs for CD56+ large granular lymphocytes (LGL). Conventional techniques using chromium release assays to measure lymphocyte-mediated cytotoxicity suffer from disadvantages, including radioactive contamination and the inability to simultaneously determine K-562 and/or CD56+ lymphocyte phenotypes. We illustrate here a three-color flow cytometric method providing for the simultaneous evaluation of K-562–CD56+ LGL binding, K-562 cell viability, and the status of K-562 cell differentiation. Phorbol 12-myristate 13-acetate (PMA) engenders megakaryocytic differentiation in K-562 cell populations, as measured by presentation of the β3 integrin (gpIIIa, CD61), while maintaining a negative expression of MHC-I and MHC-II molecules. Using the auto-fluorescence of K-562 cells, flow cytometry can be used to demonstrate a significant decrease in CD56+ LGL activity against K-562 cells in populations pre-incubated with PMA. The capacity of three-color flow cytometry to measure lymphocyte–target cell binding and cell death kinetics, while simultaneously determining target cell phenotype, permits the specific localization of CD61-expressing K-562 cells to areas inconsistent with CD56+ LGL-mediated patterns of lysis.