Volume 137, Issue 2, Pages 682-690 (August 2009) Profile of Tumor Antigen-Specific CD8 T Cells in Patients With Hepatitis B Virus-Related Hepatocellular Carcinoma  Adam J. Gehring, Zi Zong Ho, Anthony T. Tan, Myat Oo Aung, Kang Hoe Lee, Kai Chah Tan, Seng Gee Lim, Antonio Bertoletti  Gastroenterology  Volume 137, Issue 2, Pages 682-690 (August 2009) DOI: 10.1053/j.gastro.2009.04.045 Copyright © 2009 AGA Institute Terms and Conditions

Figure 1 Frequency of tumor antigen-specific responses. (A) Summary of the number of patients in each group with a TAA response or HBV-specific response. (B) Graph shows the number of patients within each group who had T-cell responses to individual tumor antigens. (C) Mean frequency of tumor antigen and HBV-specific T cells from each patient group. Frequency was calculated from ELISPOT results, and background from unstimulated wells was subtracted to give specific frequency. Gastroenterology 2009 137, 682-690DOI: (10.1053/j.gastro.2009.04.045) Copyright © 2009 AGA Institute Terms and Conditions

Figure 2 Dominant tumor antigen epitopes expand T cells in patients with diverse HLA-A2 subtypes. (A) Results from high-resolution HLA-A2 subtyping demonstrate a diverse HLA-A2 subtype profile among patients tested. (B) Analysis of all tumor antigen responses and HLA-A2 subtypes able to elicit T-cell responses. Gastroenterology 2009 137, 682-690DOI: (10.1053/j.gastro.2009.04.045) Copyright © 2009 AGA Institute Terms and Conditions

Figure 3 Affinity and function of tumor antigen-specific T cells. HCC (A) and cirrhosis (B) patient TAA-specific T-cell affinity was tested by stimulating tumor antigen-specific T cells with HLA-A2+ T2 cells. Tumor antigen affinity was compared with an HBV core-specific T-cell clone known to efficiently recognize antigen presented by hepatocytes. Values presented as percentage maximum response because each T-cell line differed in frequency and HBV core T-cell clone is 100% specific. Function of T cells from HCC (C) or cirrhosis (D) patients. Cells stained with CD8, IFN-γ (top row), CD107a (middle row), and TNF-α (bottom row). Data presented are representative patients from each group. Gastroenterology 2009 137, 682-690DOI: (10.1053/j.gastro.2009.04.045) Copyright © 2009 AGA Institute Terms and Conditions

Figure 4 (A) PD-1 staining on tumor and nontumor T cells from HCC patients. Intrahepatic lymphocytes (IHL, nontumor T cells), tumor-infiltrating T cells (TIL), and PBMC from HCC/HBV or chronic HBV patients were isolated and stained with CD3 and PD-1. Data presented as percentage PD-1+ CD3+ cells. (B) Fold increase in TAA-specific T-cell frequency after expansion in the presence of anti-PD-L1 blocking antibody. (C) Tumor antigen T-cell expansion in presence of anti-PD-L1 blocking antibody. Function of peptide-specific T cells was tested by staining for IFN-γ and TNF-α. Data presented are from 1 patient and representative of 5 epitope-specific responses from 4 individual HCC patients. Gastroenterology 2009 137, 682-690DOI: (10.1053/j.gastro.2009.04.045) Copyright © 2009 AGA Institute Terms and Conditions