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C. Yu, A. Amadi-Obi, R. Mahdi, X. Liu, C. Egwuagu; Daclizumab Blocks Expansion Of ThIL17 Cells in Human Peripheral Blood Mononuclear Cells (PBMC). Invest. Ophthalmol. Vis. Sci. 2007;48(13):5182.
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© ARVO (1962-2015); The Authors (2016-present)
Uveitis is a T-cell mediated intraocular inflammatory disease of presumed autoimmune etiology. Although administering IFNα//ß or inhibiting IL-2 receptor α subunit by Daclizumab is a partially effective therapy, pathogenic T-cell subtypes that cause uveitis or mechanism of IL-2 or IFNα//ß actions remain unknown. Here, we have used PBMC/CD4+ T cells from normal human donors to examine potential role of IL-2 in expansion of ThIL17 cells, a newly described pathogenic T-helper subtype implicated in multiple sclerosis, inflammatory bowel disease and arthritis.
Human PBMC was isolated from buffy coats by density gradient centrifugation. Human CD4+ T cells were enriched (98% purity) by RosettSep Cocktail (StemCell Tec.). PBMC and CD4+ T cells were cultured in medium containing various combinations of IL-2, IL-7, IL-15, IL-23, TGF-ß1/IL-6, anti-CD3/-CD28 Abs or Daclizumab for 4 days. Cytokine analysis was performed by ELISA or intracellular staining and surface phenotype by FACS analysis.
We showed: (i) Exposure of normal human PBMC/CD4+ T cells to exogenous IL-2 induces significant expansion of ThIL17 cells; (ii) Analysis of NK cells, monocytes, polymorphonuclear cells, and B-cells, reveal that T cells are the sole source of IL-17 in human PBMC; (iii) Among T cell subsets examined, including, CD8+, CD25, CD45RO+ or CD45RA+, CD62L, CD69 CD4+ and γ/Δ T-cells, IL-2-induced increase in IL-17 expression is primarily in CD4+ CD45RO+ CD25+population; (iv) TGF-ß1 and IL-6 or IL-23 alone is not as efficient as IL-2 in expanding ThIL17 cells in CD4+ T cells; (v) The IL-2-induced expansion is blocked by Daclizumab.
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