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The Journal of Immunology, Vol 150, Issue 4 1524-1534, Copyright © 1993 by American Association of Immunologists
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K Sebok, D Woodside, A al-Aoukaty, AD Ho, S Gluck and AA Maghazachi
Northeastern Ontario Regional Cancer Centre, Sudbury, Ontario, Canada.
The effect of IL-8 on the in vitro locomotion of human IL-2-activated natural killer (IANK) cells was studied. It was observed that IL-8 induces chemokinesis in these cells, as determined by their migration in modified Boyden chambers. Bacterial toxins such as cholera toxin or pertussis toxin inhibited IL-8-induced chemokinetic activity, suggesting the involvement of guanine nucleotide-binding (G) proteins in IL-8 signal transduction in these cells. Pertussis toxin ADP- ribosylates a 39-kDa protein, whereas cholera toxin ADP-ribosylates a 43- to 45-kDa protein. Pretreatment of IANK cell membranes with 0.01 or 0.1 ng/ml of IL-8 and/or 5 microM GTP-gamma S did not affect pertussis toxin- or cholera toxin-dependent ADP-ribosylation. Western blot analysis showed that IANK cell membranes possess one Gi (39 kDa), two Gs (43 kDa and 45 kDa), and one Go (39 kDa). Pretreatment of IANK cell membranes with concentrations between 0.001 to 1.0 ng/ml of IL-8 resulted in the disappearance of the 39 kDa Go, but not Gi or Gs protein(s), suggesting that IL-8 receptors expressed on IANK cells are coupled to Go. Various concentrations of IL-8 enhanced the binding of GTP-gamma 35 S to IANK cell membranes, which further indicates the coupling of G proteins to IL-8 receptors in IANK cells.
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