Table 3.

Effect of CADO and adenosine on cytokine production by LAK cells stimulated by cross-linking of Ly49D

GroupsIFN-γGM-CSFMIP-1αTNF-α
Experiment 1
    Control308 ± 51822 ± 304,406 ± 1,239224 ± 5
    Ly49D968 ± 3022,467 ± 40911,942 ± 958497 ± 68
    Ly49D + CADO94 ± 17420 ± 541,195 ± 71129 ± 15
    Ly49D + ZM241385 + CADO537 ± 341,426 ± 2459,712 ± 3,109296 ± 21
Experiment 2
    Control612 ± 24407 ± 13862 ± 349124 ± 7
    Ly49D836 ± 141606 ± 418,195 ± 77161 ± 13
    Ly49D + adenosine + EHNA257 ± 2283 ± 24738 ± 300101 ± 11
    Ly49D + EHNA640 ± 9399 ± 164,135 ± 459139 ± 16
  • NOTE: Plastic adherent purified LAK cells were washed and rested for 2 hours in the absence of IL-2. LAK cells were incubated with test chemicals in tubes for 20 minutes, then anti-Ly49D rat mAb was added (1 μg/well) and LAK cells were plated into 96-well plates precoated with 2 μg/well rabbit anti-rat IgG (0.5 × 106 per well). After 6 hours of incubation at 37°C, cytokine production was analyzed using a multiplex kit and Luminex LabMAP technology. Anti-Ly49D mAb significantly increased cytokine production (P < 0.05) and CADO or ADO + EHNA significantly attenuated this response (P < 0.05 and P < 0.01, respectively). The inhibitory effect of CADO was significantly reversed by ZM241385 (P < 0.05; experiment 1). The differences between groups EHNA alone and ADO + EHNA are significant (P < 0.05; experiment 2). The experiments were repeated twice.