title: An Altered gp100 Peptide Ligand with Decreased Binding by TCR and CD8α Dissects T Cell Cytotoxicity from Production of Cytokines and Activation of NFAT.
abstract: Altered peptide ligands (APLs) provide useful tools to study T cell activation and potentially direct immune responses to improve treatment of cancer patients. To better understand and exploit APLs, we studied the relationship between APLs and T cell function in more detail. Here, we tested a broad panel of gp100280-288 APLs with respect to T cell cytotoxicity, production of cytokines, and activation of Nuclear Factor of Activated T cells (NFAT) by human T cells gene-engineered with a gp100-HLA-A2-specific TCRαβ. We demonstrated that gp100-specific cytotoxicity, production of cytokines, and activation of NFAT were not affected by APLs with single amino acid substitutions, except for an APL with an amino acid substitution at position 3 (APL A3), which did not elicit any T cell response. A gp100 peptide with a double amino acid mutation (APL S4S6) elicited T cell cytotoxicity and production of IFNγ, and to a lesser extent TNFα, IL-4, and IL-5, but not production of IL-2 and IL-10, or activation of NFAT. Notably, T cell receptor (TCR)-mediated functions showed decreases in sensitivities for S4S6 versus gp100 wild-type (wt) peptide, which were minor for cytotoxicity but at least a 1000-fold more prominent for the production of cytokines. TCR-engineered T cells did not bind A3-HLA-A2, but did bind S4S6-HLA-A2 although to a lowered extent compared to wt peptide-HLA-A2. Moreover, S4S6-induced T cell function demonstrated an enhanced dependency on CD8α. Taken together, most gp100 APLs functioned as agonists, but A3 and S4S6 peptides acted as a null ligand and partial agonist, respectively. Our results further suggest that TCR-mediated cytotoxicity can be dissected from production of cytokines and activation of NFAT, and that the agonist potential of peptide mutants relates to the extent of binding by TCR and CD8α. These findings may facilitate the design of APLs to advance the study of T cell activation and their use for therapeutic applications.
authors.affiliation: Laboratory of Experimental Tumor Immunology, Department Medical Oncology, Erasmus MC Cancer Institute , Rotterdam , Netherlands.
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
authors.forename: Miriam
authors.initials: M
authors.lastname: Coccoris
authors.forename: Joost
authors.initials: J
authors.lastname: Drexhage
authors.forename: Christiaan
authors.initials: C
authors.lastname: Knoop
authors.forename: I Jolanda M
authors.initials: IJ
authors.lastname: de Vries
authors.forename: Gosse J
authors.initials: GJ
authors.lastname: Adema
authors.forename: Reno
authors.initials: R
authors.lastname: Debets
keywords: T cell receptor; activation of nuclear factor of activated T cells; altered peptide ligands; cytokine production; cytotoxicity; human T lymphocytes
references.citation: Immunol Today. 1991 Nov;12(11):415-9
references.pmid: 1786074
references.citation: Cancer Res. 2003 Apr 1;63(7):1560-7
references.pmid: 12670905
references.citation: J Immunol. 2003 Feb 15;170(4):2186-94
references.pmid: 12574392
references.citation: Nature. 1996 Apr 11;380(6574):495-8
references.pmid: 8606766
references.citation: Semin Immunol. 1996 Apr;8(2):83-101
references.pmid: 8920243
references.citation: Cancer Res. 1995 Nov 15;55(22):5330-4
references.pmid: 7585596
references.citation: Scand J Immunol. 2006 Dec;64(6):581-7
references.pmid: 17083613
references.citation: Immunology. 2012 Oct;137(2):139-48
references.pmid: 22804746
references.citation: Life Sci. 1999;64(9):717-31
references.pmid: 10075104
references.citation: J Immunol. 1999 Sep 1;163(5):2601-9
references.pmid: 10452999
references.citation: Hum Immunol. 1995 Dec;44(4):189-98
references.pmid: 8770631
references.citation: Int Immunol. 2006 Apr;18(4):591-601
references.pmid: 16507598
references.citation: J Immunol. 2000 Oct 15;165(8):4264-71
references.pmid: 11035060
references.citation: Oncoimmunology. 2013 Feb 1;2(2):e22679
references.pmid: 23524444
references.citation: J Exp Med. 1998 May 18;187(10):1699-709
references.pmid: 9584148
references.citation: Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6458-62
references.pmid: 8022805
references.citation: Cancer Gene Ther. 2006 May;13(5):503-9
references.pmid: 16282986
references.citation: J Clin Oncol. 2011 Mar 1;29(7):917-24
references.pmid: 21282551
references.citation: J Immunol. 2003 Feb 1;170(3):1209-17
references.pmid: 12538678
references.citation: Expert Opin Biol Ther. 2008 Dec;8(12):1873-84
references.pmid: 18990075
references.citation: Hum Gene Ther Methods. 2012 Jun;23(3):213-24
references.pmid: 22871260
references.citation: Gene Ther. 1998 Sep;5(9):1195-203
references.pmid: 9930320
references.citation: Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):4036-40
references.pmid: 7732026
references.citation: J Immunol. 2010 Aug 15;185(4):2600-10
references.pmid: 20639478
references.citation: J Exp Med. 1996 Apr 1;183(4):1917-21
references.pmid: 8666949
references.citation: J Biol Chem. 2007 Aug 17;282(33):23799-810
references.pmid: 17540778
references.citation: Eur J Immunol. 1997 Sep;27(9):2195-203
references.pmid: 9341759
references.citation: J Immunol. 2000 Jul 15;165(2):948-55
references.pmid: 10878370
references.citation: J Immunol Methods. 2003 Sep;280(1-2):13-24
references.pmid: 12972184
references.citation: J Immunol. 2003 Sep 15;171(6):3287-95
references.pmid: 12960359
references.citation: J Immunol. 1998 Feb 15;160(4):1750-8
references.pmid: 9469433
references.citation: J Immunother Emphasis Tumor Immunol. 1993 Aug;14(2):121-6
references.pmid: 7506574
references.citation: Blood. 2009 Jul 16;114(3):535-46
references.pmid: 19451549
references.citation: Cancer Immunol Immunother. 2011 Feb;60(2):249-60
references.pmid: 21069321
references.citation: Cancer Immunol Immunother. 2007 Mar;56(3):319-29
references.pmid: 16906394
references.citation: Expert Opin Biol Ther. 2010 Apr;10(4):547-62
references.pmid: 20146634
references.citation: Sci Transl Med. 2012 Mar 28;4(127):127ps8
references.pmid: 22461638
references.citation: J Immunol. 2006 Jul 15;177(2):991-8
references.pmid: 16818755
references.citation: Cancer Immunol Immunother. 2007 Mar;56(3):287-301
references.pmid: 16874487
references.citation: J Immunol. 1992 Dec 1;149(11):3580-7
references.pmid: 1331239
references.citation: J Immunol. 2001 Jul 15;167(2):787-96
references.pmid: 11441084
references.citation: J Immunol. 1999 Feb 15;162(4):2073-80
references.pmid: 9973480
references.citation: Curr Opin Immunol. 2002 Feb;14(1):66-74
references.pmid: 11790534
references.citation: Clin Cancer Res. 2002 Jan;8(1):41-53
references.pmid: 11801539
references.citation: J Exp Med. 1994 Mar 1;179(3):1005-9
references.pmid: 8113668
references.citation: Biochem Biophys Res Commun. 2006 Mar 3;341(1):19-27
references.pmid: 16412387
references.citation: J Immunol. 1996 Sep 15;157(6):2539-48
references.pmid: 8805655
references.citation: Int J Cancer. 2001 Nov;94(4):540-4
references.pmid: 11745441
references.citation: Proc Natl Acad Sci U S A. 2008 Mar 11;105(10):3849-54
references.pmid: 18319339
references.citation: Annu Rev Immunol. 1996;14:1-27
references.pmid: 8717505
references.citation: Cancer Immunol Immunother. 2006 Nov;55(11):1358-66
references.pmid: 16435129
references.citation: J Immunol. 1998 Jun 15;160(12):5807-14
references.pmid: 9637491