Figure 6. SDF-1/CXCL12 and TARC/CCL17 gene and protein production in cultured human conjunctival fibroblasts (HCFs)

A: Cultured conjunctival fibroblasts were homogenized in Isogen®. Water was used as a negative control. The TARC/CCL17 and SDF-1/CXCL12 genes are expressed in cultured conjunctival fibroblasts. Lane M shows a molecular weight marker (100 bp ladder). Lane C: Cultured conjunctival fibroblasts. Lane NC: Negative control. B: Cultured conjunctival fibroblasts were incubated in serum free medium for 24 h with IFN-γ (10 ng/ml), IL-4 (10 ng/ml), IL-13 (10 ng/ml), TNF-α (10 ng/ml), IFN-γ+TNF-α (10 ng/ml each), or the vehicle only. The SDF-1/CXCL12 and TARC/CCL17 concentrations in the culture supernatant were measured by ELISA. A significantly large amount of SDF-1/CXCL12 is produced in HCFs stimulated with recombinant IFN-γ, but not with IL-4, IL-13, or TNF-α in comparison with the control samples. The high SDF-1/CXCL12 concentration produced with IFN-γ is decreased in the presence of the same amount of recombinant TNF-α. The TARC/CCL17 concentration was undetectable with these cytokine stimulations (data not shown). Values presented are means; the error bars represent the standard deviations. The single asterisk indicates a statistical difference between IFN-γ and control (p=0.0057; Fisher's protected least significant difference test); the double asterisk indicates a statistical difference between IFN-γ+TNF-α and IFN-γ (p=0.0115; Fisher's protected least significant difference test). Similar results were obtained for a different set of culture dishes.