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Regulation of natural antimicrobial defences in human vaginal epithelial cells
  1. C Foster1,2,
  2. E Chin-Smith1,
  3. R Tribe1
  1. 1Division of Women's Health, King's College London and King's Health Partners, London, UK
  2. 2Nursing and Health Sciences, Monash University, Melbourne, Australia

Abstract

Background The involvement of natural antimicrobial peptides in infection-associated preterm birth is unknown. Our aim was to assess regulation by granulocyte-macrophage colony-stimulating factor, monocyte-chemotactic-protein-1 (GM-CSF and MCP-1, interleukin-1 (IL) and lipopolysaccharide (LPS) in cultured human vaginal epithelial cells.

Methods VK2/E6E7 cells were treated with IL-1, MCP-1 and GM-CSF (20/100/200/500 pg/ml and 1 ng/ml) at 2 h, 4 h, 6 h and 24 h (n=4). Cells (n=3–4) were also treated with MCP-1, IL-1 (1 ng/ml or 10 ng/ml) or LPS (50 ng/ml or 100 ng/ml) for 6 h or 24 h. mRNA was extracted and assessed SLPI, SKALP, HBD1 and HBD2 by qPCR. Data was analysed by ANOVA with repeated measures.

Results SLPI, SKALP and HBD1–2 transcripts were identified in VK2/E6E7 cells. 100 ng/ml IL-1 increased SKALP mRNA expression at 2 h (p=0.005) and 1 ng/ml at 4 h and 6 h (p=0.0003).Expression was maintained at 24 h with 10 ng/ml (p=0.0002). GM-CSF (100 pg/ml) suppressed SKALP mRNA expression after 24 h (p=0.005). 1 ng/ml and 10 ng/ml IL-1 increased HBD2 mRNA expression at 6 h (p=0.003) being maintained at 24 h with 10 ng/ml IL-1 (p=0.0002). LPS (100 ng/ml) increased SKALP mRNA expression at 6 h and 24 h (p<0.05).

Conclusions This data shows NAs expression and regulation by inflammatory agents in VK2/E6E7 cells. GM-CSF suppression of SKALP, at concentrations found in cervico-vaginal fluid, provides a novel mechanism by which impaired immune defences against ascending infection in women at risk of preterm labour.

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Footnotes

  • Funding Tommy's Charity (1060508).

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