Abstract
The proliferation of CD3-LAK cells could be induced from normal peripheral blood mononuclear cells (PBMC) after in vitro treatment with CD2 monoclonal antibody (MoAb) plus rabbit complement and culture with IL-1 plus IL-2. This presentation exhibited the results of the cloning of the LAK cells.
PBMC from healthy donors were incubated with CD2 MoAb plus rabbit complement for 1 hr, then washed with HBSS and cultured with rIL-1β (0.5μg/dl) plus rIL-2 (2μg/dl) in RPMI-1640 culture medium. Two to ten days later, the LAK cells were cloned under limiting conditions in 96-well microtiter plates. Ten to 14 days later the cloned cells were pipetted into a culture flask and continued to culture with the addition of rIL-1 plus rIL-2. After the cloned cells increased to sufficient numbers of cells, their surface markers were analyzed using various MoAb by FCM and histostaining techniques, and the non-major histocompatibility-restricted cytotoxicity (NMRC) was tested against target K562 cells.
The results showed 112 clones were WT31+γ/δ-1-, 62 clones were WT31-γ/δ-1+ and 101 clones were WT31-γ/δ-. Most of the WT31-γ/δ-1- clones showed CD2+, CD7+ and CD56+, but no CD3, βF 1 and TCRδ-1 on their cell surfaces. Some of the WT31-γ/δ-1- clones showed CD2- and CD56-. The NMRC showed 13 clones out of 27 WT31-γ/δ-1- clones tested had a strong cytotoxicity to K562 target cells but none of the WT31+ nor γ/β+ clones had.
The surface markers of 10 out of 13 NMRC positive clones showed both CD2 and CD56 were positive. From these results, CD2 and CD56 antigens may have some important effect on CD3- LAK cells regarding to the NMRC activity.
