Expression of a variant of CD28 on a subpopulation of human NK cells: Implications for B7-mediated stimulation of NK cells

The ability of NK cells to kill tumor cells is controlled by a balance betw een activating and inhibitory signals transduced by distinct receptors, In murine tumor models, the costimulatory molecule B7.1 not only acts as a pos itive trigger for NK-mediated cytotoxicity but can also overcome negative s ignaling transduced by MHC class I molecules. In this study, we have evalua ted the potential of human B7.1-CD28 interaction as an activating trigger f or human blood NK cells. Using multiparameter flow cytometric analysis and a panel of different CD28 mAbs, we show that human peripheral blood NK cell s (defined by CD56(+), CD16(+), and CD3(-) surface expression) express the CD28 costimulatory receptor, with its detection totally dependent on the mA b used. In addition, the level of CD28 varies among individuals and on diff erent NK cell lines, irrespective of CD28 steady-state mRNA levels. By perf orming Ab binding studies on T cells, our data strongly suggest that bindin g of two of the anti-CD28 Abs (clones 9.3 and CD28.2) is to a different epi tope to that recognized by clones L293 and YTH913.12, which is perhaps modi fied in the CD28 molecule expressed by the NK cells. We also show that B7.1 enhances the NK-mediated lysis of NK-sensitive but not of NK-resistant tum or cells and that this increased lysis is dependent on CD28-B7 interactions as shown by the ability of Abs to block this lysis. Coculture of the B7.1- positive NK-sensitive cells also led to the activation of the NK cells, as determined by the expression of CD69, CD25, and HLA class II.