Upregulated Expression of Indoleamine 2, 3-Dioxygenase in Primary Breast Cancer Correlates with Increase of Infiltrated Regulatory T Cells In Situ and Lymph Node Metastasis
Figure 4
Foxp3 expression in CD3+ T cells was upregulated both at mRNA and protein levels after coculture with IDO/CHO cells. The expression of Foxp3 gene at mRNA and protein levels in treated and untreated T cells were detected using qRT-PCR assay and Western Blot method. (a) After 7 day coculture, the relative mRNA level of Foxp3 in the CD3+ T cells treated with the CHO/IDO cells was significantly higher than that in the CD3+ T cells treated with the CHO/EGFP control cells or untreated CD3+ T cells (lane 1: β-actin in the CD3+ T control cells; lane 2: Foxp3 in the CD3+ T control cells; lane 3: β-actin in the T cells treated with CHO/IDO cells; lane 4: Foxp3 in the T cells treated with CHO/IDO cells; lane 5: β-actin in the T cells treated with CHO/EGFP cells; lane 6: Foxp3 in the T cells treated with CHO/EGFP cells; lane 7: DL2000 Marker. (b) The mRNA amount of Foxp3 in the CD3+ T cells stimulated by the CHO/IDO cells was significantly higher than that in the CD3+ T cells stimulated by the CHO/EGFP cells and the unstimulated CD3+ T cells control. (c) Foxp3 expression at protein level in treated and untreated T cells detected by Western Blot analysis. Foxp3 expression was exclusively detected in the cell lysates of CD3+ T cells treated with CHO/IDO cells, indicating a 48 kD protein band reactive to a Foxp3-specific monoclonal antibody (lane 1: Foxp3 in CD3+ T cells treated with CHO/IDO cells; lane 2: Foxp3 in CD3+ T cells treated with CHO/EGFP cells; lane 3: Foxp3 in control CD3+ T cells).
