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. 2012 Feb;3(2):231-236.
doi: 10.3892/etm.2011.402. Epub 2011 Dec 1.

Influence of interleukin-8 (IL-8) and IL-8 receptors on the migration of human keratinocytes, the role of PLC-γ and potential clinical implications

Wen G Jiang  1 , Andrew J Sanders, Fiona Ruge, Keith G Harding
Affiliations

Influence of interleukin-8 (IL-8) and IL-8 receptors on the migration of human keratinocytes, the role of PLC-γ and potential clinical implications

Wen G Jiang et al. Exp Ther Med. 2012 Feb.

Abstract

Interleukin (IL)-8 is a pro-inflammatory cytokine that has a direct effect on immune cells, including polymorphonuclear cells. Keratinocytes are a rich source of IL-8. However, there is little knowledge on the role of IL-8 in clinical wound healing and the direct biological effect of IL-8 on keratinocytes. In this study, the effect of recombinant human IL-8 (rhIL-8) on migration and adhesion was tested using HaCaT keratinocytes as a cell model. The cell functions were evaluated using impedance cell sensing. The expression of IL-8 receptor (IL-8R) transcripts in human skin and wounds (acute and chronic) was assessed using real-time transcript analysis. rhIL-8 significantly increased the migration of keratinocytes (3.5±0.3 for cells treated with IL-8 vs. 2.7±0.6 for controls; p=0.029). It is interesting to note that treatment of keratinocytes with IL-8 resulted in a marked shift in the responsive frequencies. IL-8 only resulted in a marginal increase in cell adhesion, which was particularly noticeable at high frequencies. The PLC-γ inhibitor completely eradicated the action of IL-8 on the migration of HaCaT cells. Using real time PCR, it was found that chronic wounds had significantly lower levels of the B form of the IL-8R (IL-8RB) (p=0.045) and marginally lower levels of the A form, IL-8RA, in comparison with acute wounds. Therefore, IL-8 has a direct and profound stimulatory effect on the migration of human keratinocytes, which is likely to occur via the PLC-γ pathway. Together with a reduced level of IL-8Rs in difficult-healing wounds, IL-8 has a clear prognostic and therapeutic value in wound healing.

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Figures

Figure 1.
Figure 1.
The effect of rhIL-8 on the migration of HaCaT cells. Confluent HaCaT cells in the 96W1E array were wounded for 20 sec at 6 V, following which the migration was immediately traced for up to 8 h over 12 frequencies [shown are (A) 1000, (B) 4000 and (C) 32000 Hz]. rhIL-8 markedly increased the pace of migration of the cells. rhIL-8, recombinant human IL-8.
Figure 2.
Figure 2.
Effect of rhIL-8 on cellular migration of HaCaT cells, a 3-D demonstration. Cells were wounded in a similar fashion as in Fig. 1 and recorded following wounding over 7 h. When treated with IL-8, cells migrated earlier (B) and faster than the control cells (B). (A and B) mean of 8 traces, (C) Cell modelling. *p<0.01 vs. control cells. rhIL-8, recombinant human IL-8.
Figure 3.
Figure 3.
rhIL-8 had a significant impact on the adhesion of keratinocytes, assessed using the ECIS method. Cells were added to the ECIS arrays, pre-treated with 10% FCS; the adhesion was traced immediately following addition of the cells. (A) Traces of adhesion, (B) normalised resistance of adhesion traces, (C and D) 3-D modelling of adhesion (as resistance). rhIL-8, recombinant human IL-8; ECIS, electric cell-substrate impedance sensing; FCS, fetal calf serum; WT, wild-type.
Figure 4.
Figure 4.
Potential signalling pathways involved in IL-8-mediated migration in HaCaT cells. (A) ECIS traces following electric wounding, (B) Mean changes of resistance. *p<0.05 vs. without IL-8. n=4. rhIL-8, recombinant human IL-8; ECIS, electric cell-substrate impedance sensing.
Figure 5.
Figure 5.
The levels of IL-8 receptor transcript expression in acute and chronic wounds. (A) IL-RA, (B) IL-8RB. The levels shown are the medians. *p<0.05 vs. acute wound tissues, as shown by the Mann-Whitney U test. IL-8R, IL-8 receptor.
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References

    1. Van Damme J, Decock B, Conings R, Lenaerts JP, Opdenakker G, Billiau A. The chemotactic activity for granulocytes produced by virally infected fibroblasts is identical to monocyte-derived interleukin 8. Eur J Immunol. 1989;19:1189–1194. - PubMed
    1. Wolpe SD, Cerami A. Macrophage inflammatory proteins 1 and 2: members of a novel superfamily of cytokines. FASEB J. 1989;3:2565–2573. - PubMed
    1. Modi WS, Chen ZQ. Localization of the human CXC chemokine subfamily on the long arm of chromosome 4 using radiation hybrids. Genomics. 1998;47:136–139. - PubMed
    1. Hull J, Thomson A, Kwiatkowski D. Association of respiratory syncytial virus bronchiolitis with the interleukin 8 gene region in UK families. Thorax. 2000;55:1023–1027. - PMC - PubMed
    1. Smyth RL, Mobbs K, O’Hea U, Ashby D, Hart CA. The association between disease severity, cytokines and virus genotype in infants with respiratory syncytial virus (RSV) bronchiolitis. Arch Dis Child. 2000;82(Suppl 1):A4–A5.

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