Co-culture of healthy human keratinocytes and T-cells promotes keratinocyte chemokine production and RORγt-positive IL-17 producing T-cell populations

J Dermatol Sci. 2013 Jan;69(1):44-53. doi: 10.1016/j.jdermsci.2012.10.004. Epub 2012 Oct 17.

Abstract

Background: Both keratinocytes and T-cells are crucial players in cutaneous immune responses. We hypothesized that direct interactions between keratinocytes and T-cell subsets could shape the nature or strength of the local immune response.

Objective: We investigated direct interactions between keratinocytes and T-cell subsets, focused on keratinocyte chemokine production and T-cell phenotype and cytokine production.

Methods: A newly developed in vitro serum free co-culture model using primary keratinocytes and T-cells subsets from healthy human donors was used. Keratinocyte chemokine production was analyzed with luminex, T-cell phenotype and cytokine production were analyzed with flow cytometry.

Results: Our data show that upon co-culture with CD4(pos) or CD8(pos) T-cells primary human keratinocytes increased production of functionally active chemokines CCL2, CCL20 and CXCL10 and that regulatory T-cells did not regulate keratinocyte chemokine production. Next to that, we found that keratinocytes skewed CD4(pos) and CD8(pos) T-cell populations toward an IL-17(pos) CCR6(pos) RORγt(pos) phenotype in a cell-cell contact independent manner, and that Treg were able to decrease the absolute number of IL-17 producing T-cells in keratinocyte/T-cell co-cultures. Correspondingly, freshly isolated skin-derived T-cell populations contained relatively high percentages of IL-17(pos) cells.

Conclusion: We provide evidence that keratinocyte/T-cell communication may regulate leukocyte influx in the skin, and that keratinocytes enrich T-cell populations for Th17/Tc17 cells. Accumulation of Th17/Tc17 cells, but not chemokine production, appears under the control of regulatory T-cells. Dysregulation of these processes may well contribute to the pathophysiology of inflammatory skin diseases.

MeSH terms

  • Analysis of Variance
  • CD4-Positive T-Lymphocytes / immunology*
  • CD8-Positive T-Lymphocytes / immunology*
  • Cell Communication
  • Cells, Cultured
  • Chemokine CCL2 / metabolism
  • Chemokine CCL20 / metabolism
  • Chemokine CXCL10 / metabolism
  • Coculture Techniques
  • Fibroblasts / immunology
  • Humans
  • Keratinocytes / immunology*
  • Keratinocytes / metabolism*
  • Nuclear Receptor Subfamily 1, Group F, Member 3
  • Phenotype
  • Skin / immunology
  • T-Lymphocytes, Regulatory / immunology
  • Th17 Cells / immunology
  • Th17 Cells / metabolism

Substances

  • Chemokine CCL2
  • Chemokine CCL20
  • Chemokine CXCL10
  • Nuclear Receptor Subfamily 1, Group F, Member 3