title: Aspartate-β-hydroxylase induces epitope-specific T cell responses in hepatocellular carcinoma.
abstract: Hepatocellular carcinoma (HCC) has a poor prognosis due to high recurrence rate. Aspartate-β-hydroxylase (ASPH) is a highly conserved transmembrane protein, which is over expressed in HCC and promotes a malignant phenotype. The capability of ASPH protein-derived HLA class I and II peptides to generate antigen specific CD4(+) and CD8(+) immune responses is unknown. Therefore, these studies aim to define the epitope specific components required for a peptide based candidate vaccine. Monocyte-derived dendritic cells (DCs) generated from the peripheral blood mononuclear cells (PBMCs) of HCC patients were loaded with ASPH protein. Helper CD4(+) T cells and CD8(+) cytotoxic T lymphocytes (CTLs) were co-incubated with the DCs; T cell activation was evaluated by flow cytometric analysis. Immunoinformatics tools were used to predict HLA class I- and class II-restricted ASPH sequences, and the corresponding peptides were synthesized. The immunogenicity of each peptide in cultures of human PBMCs was determined by IFN-γ ELISpot assay. ASPH protein-loaded DCs activated both CD4(+) and CD8(+) T cells contained within the PBMC population derived from HCC patients. Furthermore, the predicted HLA class I- and class II-restricted ASPH peptides were significantly immunogenic. Both HLA class I- and class II-restricted peptides derived from ASPH induce T cell activation in HCC. We observed that ASPH protein and related peptides were highly immunogenic in patients with HCC and produce the type of cellular immune responses required for generation of anti-tumor activity.
authors: Liver Research Center, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, RI 02903, USA.
authors: Yoshito
authors: Y
authors: Tomimaru
authors: Department of Medicine, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, RI 02903, USA.
authors: Sasmita
authors: S
authors: Mishra
authors: Department of Medicine, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, RI 02903, USA.
authors: Howard
authors: H
authors: Safran
authors: Department of Surgery, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, RI 02903, USA.
authors: Kevin P
authors: KP
authors: Charpentier
authors: EpiVax, Inc., Providence, RI 02903, USA.
authors: William
authors: W
authors: Martin
authors: EpiVax, Inc., Providence, RI 02903, USA.
authors: Anne S
authors: AS
authors: De Groot
authors: Department of Medicine, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, RI 02903, USA.
authors: Stephen H
authors: SH
authors: Gregory
authors: Liver Research Center, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, RI 02903, USA; Department of Medicine, Rhode Island Hospital and the Warren Alpert Medical School of Brown University, Providence, RI 02903, USA. Electronic address: jack_wands_md@brown.edu.
authors: Jack R
authors: JR
authors: Wands
mesh_terms: D002135:Calcium-Binding Proteins / Q000378:metabolism*; D019496:Cancer Vaccines / Q000276:immunology; D006528:Carcinoma, Hepatocellular / Q000201:enzymology / Q000276:immunology*; D003713:Dendritic Cells / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D015789:HLA-A2 Antigen / Q000276:immunology; D059811:HLA-DRB1 Chains / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008113:Liver Neoplasms / Q000201:enzymology / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology*; D008565:Membrane Proteins / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009124:Muscle Proteins / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D022223:Vaccines, Subunit / Q000276:immunology
chemical_list: D002135:Calcium-Binding Proteins; D019496:Cancer Vaccines; D018984:Epitopes, T-Lymphocyte; D015789:HLA-A2 Antigen; D059811:HLA-DRB1 Chains; D008565:Membrane Proteins; D009124:Muscle Proteins; D022223:Vaccines, Subunit; D007371:Interferon-gamma; D006899:Mixed Function Oxygenases; C419389:ASPH protein, human
keywords: Aspartate-β-hydroxylase; Epitope; Hepatocellular carcinoma; Immunotherapy; T cells
references: J Immunol. 2006 Jul 1;177(1):712-21
references: 16785570
references: Clin Cancer Res. 2003 Dec 1;9(16 Pt 1):5902-8
references: 14676113
references: J Immunol. 2007 Feb 1;178(3):1914-22
references: 17237442
references: Gastroenterology. 2007 Jun;132(7):2557-76
references: 17570226
references: Blood. 2007 Jul 1;110(1):201-10
references: 17371945
references: Nat Rev Immunol. 2008 Jan;8(1):59-73
references: 18097448
references: Immunol Rev. 2008 Apr;222:129-44
references: 18363998
references: Cancer Sci. 2008 Jul;99(7):1420-6
references: 18422751
references: Gastroenterology. 2008 Jul;135(1):234-43
references: 18485901
references: Hepatology. 2008 Dec;48(6):1821-33
references: 19003875
references: Hepatology. 2009 Jan;49(1):124-32
references: 18980227
references: Hepatology. 2009 May;49(5):1492-502
references: 19206147
references: Dig Dis. 2009;27(2):80-92
references: 19546545
references: Cancer Sci. 2009 Oct;100(10):1935-42
references: 19604246
references: J Immunol Methods. 2010 Feb 28;353(1-2):8-19
references: 19925804
references: Liver Int. 2010 Apr;30(4):521-6
references: 20040052
references: Hepatology. 2010 Jul;52(1):164-73
references: 20578260
references: Int J Cancer. 2011 Feb 15;128(4):887-96
references: 20473887
references: J Hepatol. 2011 Apr;54(4):830-4
references: 21145836
references: Int J Oncol. 2012 Jan;40(1):63-70
references: 21922136
references: Hepatology. 2012 Jan;55(1):86-97
references: 21898484
references: J Immunol. 2000 Dec 1;165(11):6047-55
references: 11086036
references: Blood. 2000 Mar 15;95(6):2024-30
references: 10706870
references: Z Gastroenterol. 2006 Jan;44(1):43-9
references: 16397839
references: Cancer Res. 2006 Feb 15;66(4):2414-22
references: 16489048
references: Nat Rev Immunol. 2006 Apr;6(4):295-307
references: 16557261
references: J Hepatol. 2006 May;44(5):971-83
references: 16564107
references: Clin Cancer Res. 2006 May 1;12(9):2689-97
references: 16675560
references: Clin Cancer Res. 2006 May 1;12(9):2817-25
references: 16675576
references: Hepatology. 2006 Jun;43(6):1284-94
references: 16729333
references: J Hepatol. 2012 May;56(5):1129-35
references: 22245894
references: Int J Cancer. 2014 Jan 15;134(2):319-31
references: 23818246
references: Vaccine. 2014 May 30;32(26):3285-92
references: 24721533
references: Hepatology. 2014 Oct;60(4):1302-13
references: 24954865
references: Clin Cancer Res. 2004 Mar 1;10(5):1574-9
references: 15014006
references: Annu Rev Immunol. 2004;22:531-62
references: 15032588
references: Clin Cancer Res. 2004 Oct 15;10(20):6946-55
references: 15501973
references: Gastroenterology. 2004 Nov;127(5 Suppl 1):S232-41
references: 15508089
references: Proc Natl Acad Sci U S A. 1989 May;86(10):3609-13
references: 2726737
references: J Biol Chem. 1990 May 25;265(15):8558-65
references: 2187868
references: J Biol Chem. 1992 Jul 15;267(20):14322-7
references: 1378441
references: J Clin Invest. 1996 Sep 15;98(6):1313-23
references: 8823296
references: AIDS Res Hum Retroviruses. 1997 May 1;13(7):529-31
references: 9135870
references: J Immunol. 1998 Apr 1;160(7):3363-73
references: 9531296
references: Vaccine. 1998 Nov;16(19):1880-4
references: 9795396
references: Ann Surg. 1999 Jun;229(6):790-9; discussion 799-800
references: 10363892
references: Methods. 2004 Dec;34(4):476-87
references: 15542374
references: J Immunol. 2005 Feb 1;174(3):1709-16
references: 15661935
references: Blood. 2005 Apr 1;105(7):2862-8
references: 15591121
references: J Immunother. 2005 Sep-Oct;28(5):496-504
references: 16113606
references: Clin Cancer Res. 2005 Sep 15;11(18):6686-94
references: 16166448
references: Nat Med. 2005 Oct;11(10):1118-24
references: 16186818
references: J Clin Invest. 2005 Dec;115(12):3623-33
references: 16308572
references: J Immunol Methods. 2005 Dec 20;307(1-2):150-66
references: 16325197
references: Immunogenetics. 1999 Nov;50(3-4):201-12
references: 10602880
references: Oncologist. 2006 Jul-Aug;11(7):790-800
references: 16880238