Journal: PLoS ONE

Article Title: Cellular Immune Responses for Squamous Cell Carcinoma Antigen Recognized by T Cells 3 in Patients with Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0170291

Figure Lengend Snippet: (A) Immunofluorescence analysis of the expression of SART3 in hepatoma cell lines. Original magnification, ×400. (B) Immunohistochemical analysis of the expression of SART3 and alpha-fetoprotein (AFP) in sequential non-cancerous and HCC tissue sections. Original magnification, ×200 (left) and ×400 (right). (C) Analysis of SART3 expression levels among the four groups (HCC: tumor tissue in HCC patients, Non-tumor: non-tumor tissue in HCC patients, Disease control: liver tissue in disease control group, Normal liver: tissue from HCC with normal liver patients). Closed and open circles show the level of SART3 expression in the patients with liver cirrhosis and chronic hepatitis, respectively. Grayish circles show the level of SART3 expression in the patients with normal liver. (D) The expression of SART3 was also compared with AFP expression in HCC tissues.

Article Snippet: For immunofluorescence analyses, the six different hepatoma cell lines listed above and PaCa-2 (as a positive control) were fixed in acetone with methanol for 5 min and incubated with rabbit anti-human SART3 (Aviva Systems Biology, San Diego, CA, USA; 1:125) or mouse anti-human alpha-fetoprotein (AFP) (Nichirei Bioscience, Tokyo, Japan) antibody overnight at 4°C.

Techniques: Immunofluorescence, Expressing, Immunohistochemical staining, Control

Journal: PLoS ONE

Article Title: Cellular Immune Responses for Squamous Cell Carcinoma Antigen Recognized by T Cells 3 in Patients with Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0170291

Figure Lengend Snippet: (A) Interferon (IFN)-γ enzyme-linked immunospot (ELISpot) assay was performed using SART3, HIVenv 584 , and cytomegalovirus pp65 328 -derived peptides with peripheral blood mononuclear cells (PBMCs) of HCC patients. % indicates the proportion of patients who showed positive responses. (B) IFN-γ ELISpot assay in healthy donors and disease control groups. “N” denotes normal donors. “C” denotes the patients with chronic hepatitis. “L” denotes the patients with liver cirrhosis. % shows the proportion of patients who exhibited positive responses. (C) Comparison of the mean number of SART3-derived peptide-specific spots among the three groups. Error bars indicate the standard deviations. Black and white bars indicate the frequencies of SART3 109 - and SART3 315 -specific T cells, respectively.

Article Snippet: For immunofluorescence analyses, the six different hepatoma cell lines listed above and PaCa-2 (as a positive control) were fixed in acetone with methanol for 5 min and incubated with rabbit anti-human SART3 (Aviva Systems Biology, San Diego, CA, USA; 1:125) or mouse anti-human alpha-fetoprotein (AFP) (Nichirei Bioscience, Tokyo, Japan) antibody overnight at 4°C.

Techniques: Enzyme-linked Immunospot, Derivative Assay, Control, Comparison

Journal: PLoS ONE

Article Title: Cellular Immune Responses for Squamous Cell Carcinoma Antigen Recognized by T Cells 3 in Patients with Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0170291

Figure Lengend Snippet: ELISpot was performed in eight patients from whom samples of both PBMCs and TILs could be obtained. Open and solid bars show the frequency of SART3-specific T cells in PBMCs and TILs, respectively.

Article Snippet: For immunofluorescence analyses, the six different hepatoma cell lines listed above and PaCa-2 (as a positive control) were fixed in acetone with methanol for 5 min and incubated with rabbit anti-human SART3 (Aviva Systems Biology, San Diego, CA, USA; 1:125) or mouse anti-human alpha-fetoprotein (AFP) (Nichirei Bioscience, Tokyo, Japan) antibody overnight at 4°C.

Techniques: Enzyme-linked Immunospot

Journal: PLoS ONE

Article Title: Cellular Immune Responses for Squamous Cell Carcinoma Antigen Recognized by T Cells 3 in Patients with Hepatocellular Carcinoma

doi: 10.1371/journal.pone.0170291

Figure Lengend Snippet: (A) Representative results of the cytotoxic T lymphocyte (CTL) assay. The results are shown as specific cytotoxic activity, which was calculated as follows: (cytotoxic activity in the presence of peptide)–(cytotoxic activity in the absence of peptide); findings were considered positive at > 10%. (B) CTL assays [effector-to-target (E/T) ratio of 50:1] were performed in 18 HCC patients. The results are also shown as specific cytotoxic activity. Solid bars show the results for one patient. (C) Cytotoxicity of SART3-specific T-cell lines derived from the peptide was also measured against hepatoma cell lines. The cytotoxicity was considered positive when it was higher than that against K562, which shows non-specific lysis (E/T ratio of 50:1). Representative results are shown.

Article Snippet: For immunofluorescence analyses, the six different hepatoma cell lines listed above and PaCa-2 (as a positive control) were fixed in acetone with methanol for 5 min and incubated with rabbit anti-human SART3 (Aviva Systems Biology, San Diego, CA, USA; 1:125) or mouse anti-human alpha-fetoprotein (AFP) (Nichirei Bioscience, Tokyo, Japan) antibody overnight at 4°C.

Techniques: Cytotoxic T Lymphocyte Assay, Activity Assay, Derivative Assay, Lysis