Figure 2.
Figure 2. Purified NK cell populations are responsive to LFA-1–induced cytolytic activity. (A) CTLs (CD3+CD8+CD56–) or NK cells (CD3–CD8+CD56+), sorted by FACS, were cultured in IL-2 (200 ng/mL) for 12 hours and treated with either control IgG, ICAM-1, ICAM-2, or ICAM-3 (10 μg/mL) 30 minutes prior to mixing with target CMFDA-labeled HL60 at various effector-target (E/T) cell ratio (as indicated) for 4 hours. Specific cell lysis was quantitated by flow cytometry and displayed as percent specific lysis. Results are representative of 4 independent donors. Numbers indicate population frequencies. (B) CD56+CD8+CD16– and CD56+CD8+CD16+ cells, sorted by FACS, were cultured in IL-2 (200 ng/mL) for 12 hours and treated with either control IgG, ICAM-1, ICAM-2, or ICAM-3 (10 μg/mL) 30 minutes prior to mixing with target CMFDA-labeled HL60 at a particular E/T cell ratio (as indicated) for 4 hours. Specific cell lysis was quantitated by flow cytometry and displayed as percent specific lysis. (C) Cytotoxic T-cell lymphocytes (CTL: CD3+CD8+CD56–), natural killer T cells (NKT: CD3+CD8+CD56+), natural killer CD56+ cells (NK56+: CD3–CD8+CD56+), and natural killer CD56– cells (NK56–: CD3–CD8+CD56–) were sorted by FACS, cultured in IL-2 (200 ng/mL) for 12 hours, and treated with either control IgG or ICAM-2 (10 μg/mL) 30 minutes prior to mixing with target CMFDA-labeled HL60 at a 1:6.25 E/T cell ratio for 4 hours. Specific cell lysis was quantitated by flow cytometry and computed. Error bars are the standard deviation of 3 experiments.

Purified NK cell populations are responsive to LFA-1–induced cytolytic activity. (A) CTLs (CD3+CD8+CD56) or NK cells (CD3CD8+CD56+), sorted by FACS, were cultured in IL-2 (200 ng/mL) for 12 hours and treated with either control IgG, ICAM-1, ICAM-2, or ICAM-3 (10 μg/mL) 30 minutes prior to mixing with target CMFDA-labeled HL60 at various effector-target (E/T) cell ratio (as indicated) for 4 hours. Specific cell lysis was quantitated by flow cytometry and displayed as percent specific lysis. Results are representative of 4 independent donors. Numbers indicate population frequencies. (B) CD56+CD8+CD16 and CD56+CD8+CD16+ cells, sorted by FACS, were cultured in IL-2 (200 ng/mL) for 12 hours and treated with either control IgG, ICAM-1, ICAM-2, or ICAM-3 (10 μg/mL) 30 minutes prior to mixing with target CMFDA-labeled HL60 at a particular E/T cell ratio (as indicated) for 4 hours. Specific cell lysis was quantitated by flow cytometry and displayed as percent specific lysis. (C) Cytotoxic T-cell lymphocytes (CTL: CD3+CD8+CD56), natural killer T cells (NKT: CD3+CD8+CD56+), natural killer CD56+ cells (NK56+: CD3CD8+CD56+), and natural killer CD56 cells (NK56: CD3CD8+CD56) were sorted by FACS, cultured in IL-2 (200 ng/mL) for 12 hours, and treated with either control IgG or ICAM-2 (10 μg/mL) 30 minutes prior to mixing with target CMFDA-labeled HL60 at a 1:6.25 E/T cell ratio for 4 hours. Specific cell lysis was quantitated by flow cytometry and computed. Error bars are the standard deviation of 3 experiments.

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