Mode of release of interleukin‐8 from proliferating human epidermal keratinocytes in vitro

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Abstract
Keratinocytes have been shown to express interleukin-8 (IL-8) mRNA on stimulation with IL-1 and other substances. This has been assumed to account for the large amount of this neutrophil chemotactic cytokine in psoriasis. We found that, without any added agents, commercially available normal human epidermal keratinocytes proliferating in Keratinocyte Growth Medium (KGM) released a chemotactic peptide extracellularly, which was confirmed to be IL-8. To determine whether most of the IL-8 is secreted extracellularly from proliferating keratinocytes or is mainly stored to be released only on stimulation. We quantified cell-associated and released immunoreactive IL-8 from keratinocytes cultured in KGM for up to 11 days at the peptide level. The keratinocytes proliferated, taking a sigmoid growth curve, to reach a plateau at day 7. We found that the amounts of immunoreactive IL-8 gradually increased in the culture supernatant with cell growth but its prominent release took place only after the cell growth reached a plateau. The cell-associated IL-8 was much smaller in amount than that noted in the supernatant. These results suggest that IL-8 constitutively produced by keratinocytes was mostly released extracellular but that the production by actively proliferating cells seems to be far less than that by non-proliferating cells that probably occurred in an autocrine fashion under the stimulation of keratinocyte-derived cytokines accumulated in the culture medium. Neutrophil chemotactic activity assayed concomitantly showed a consistent increase during the culture period, indicating that, with their growth, the keratinocytes release substances other than IL-8 that exert an influence on neutrophil chemotactic functions.

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