Tumor necrosis factor-alpha is chemokinetic for lymphokine-activated killer cells: regulation by cyclic adenosine monophosphate.
The ability of tumor necrosis factor-alpha (TNF-alpha) to attract lymphokine-activated killer (LAK) cells in vitro was examined. Utilizing modified Boyden chambers (BC), it was observed that TNF-alpha is not chemoattractant for LAK cells. On the other hand, TNF-alpha attracted both fresh and concanavalin A-activated T cells. However, when TNF-alpha was incubated in the upper compartments of BC and in the presence of LAK cells, it enhanced the random movement of these cells across the polycarbonate membranes. The effect of TNF-alpha was inhibited by incorporating anti-TNF-alpha antibody, or a high concentration (10 ng) of TFG-beta 1. The activity of TGF-beta 1 was reversed by anti-TGF-beta 1 antibody. Cholera toxin (CT), which is known to activate the endogenous level of cyclic adenosine monophosphate (cAMP) also inhibited TNF-alpha-induced LAK cell chemokinesis. The effect of CT was mimicked by the cAMP analog dibutyryl cAMP or by the phosphodiesterase inhibitors isobutyl methylxanthine or aminophylline. Measurement of the intracellular level of cAMP showed that cells incubated for 1, 2, or 4 hr with TNF-alpha have a lower level of cAMP, whereas those incubated with a high concentration of TGF-beta 1 produced significantly higher levels of this messenger. cAMP level was also increased in cells incubated with TGF-beta 1 plus TNF-alpha. This level was reduced to the background when anti-TGF-beta 1 antibody was added to the cultures. These results suggest that cAMP negatively regulates LAK cell chemokinesis.
Maghazachi
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AA
Molecular Immunology Section, National Research Council Canada, Montréal, Québec.
D000818:Animals; D002465:Cell Movement / Q000187:drug effects*; D000242:Cyclic AMP / Q000502:physiology*; D066298:In Vitro Techniques; D015979:Killer Cells, Lymphokine-Activated / Q000502:physiology*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D051381:Rats; D011916:Rats, Inbred F344; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D010726:Phosphodiesterase Inhibitors; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D000242:Cyclic AMP