Purification and characterization of cytolytic and noncytolytic human natural killer cell subsets

Abstract

Natural killer (NK) cells form three functionally distinct populations of effectors: competent cytolytic effectors able to bind and kill target cells and two subsets of nonlytic effectors, one able and the other unable to bind target cells. A flow cytometric method was developed, based on size and two-color fluorescence of NK cell-target conjugates, for the characterization and sorting of highly purified subpopulations--killer cells, nonkiller binder cells, and free cells. Ultrastructural examination revealed that granule content was reduced in the killer cells and absent in most of the binder cells. Quantitative differences in the expression level of HLA class I, CD11b (C3bi receptor), and CD16 (receptor for the Fc portion of IgG) antigens could differentiate the subsets. The killer phenotype was HLAlo, CD11bvery hi, and CD16very lo; the binder phenotype was CD11bhi and CD16lo; and the free-cell phenotype was CD11blo and CD16hi. Cell activation was not requisite for lytic function because no difference in either expression of activation markers or cell cycle could be established among the sorted subpopulations. Although recycling function was inhibited, retention of lytic activity was enriched 4-fold in the sorted killer cell population. These results represent characterization of a successful bulk isolation of competent killer, nonkiller binder, and free cells in human NK-cell populations and should aid our understanding of NK-cell development, lineage, and function.