Figure 1

CD4+CD25+ T-cell frequency and function in PBMCs. (a) Peripheral blood CD4+CD25+ T-cell frequency in normal healthy controls (n=20), CHC infection (n=40), chronic hepatitis C with cirrhosis (n=15), and viral-related HCC (n=33). Data are expressed as box plots, in which the horizontal lines illustrate the 25th, 50th, and 75th percentiles of the frequencies of the CD4⁺CD25⁺ T lymphocytes as assessed by flow cytometry. The vertical lines represent the 10th and 90th percentiles, and circles denote values outside these percentiles. (b) Peripheral blood CD4+CD25+T-cell frequency in viral hepatitis-related HCC (n=33), non-viral-related HCC (n=7), and non-HCC liver lesions including cysts, hemangiomas, fat, and focal nodular hyperplasia (n=10). (c) FOXP3 expression in patients with viral hepatitis (n=9), and viral hepatitis-related HCC (n=12) by quantitative real-time PCR analysis. The quantity of FOXP3 mRNA is expressed relative to normal healthy controls generated using the ΔΔCt method. (d) Mean proliferative response by [³H]thymidine incorporation of CD4⁺CD25⁻ T cells in viral-related HCC (n=10) in response to PHA (5 μg/ml) when cultured alone (0:1) and on co-culture with CD4⁺CD25⁺ T cells (1:5). Isolated CD4⁺CD25⁺ cells suppressed CD4⁺CD25⁻ proliferation on co-culture and failed to display any significant proliferation when cultured alone (1:0). (e) Correlation analysis between CD4⁺CD25⁺ T-cell frequency in viral-related HCC and tumor burden (n=30). Tumor burden is defined as the following: 1—single lesion <2 cm; 2—single lesion <5 cm; 3—multiple lesions; 4—vessel involvement; 5—evidence of metastatic disease. A significant correlation is found between CD4⁺CD25⁺ T-cell frequency and tumor burden by the nonparametric Spearman rank (R=0.537; P=0.002).