title: Expanded Human Blood-Derived γδT Cells Display Potent Antigen-Presentation Functions.
abstract: Cell-based immunotherapy strategies target tumors directly (via cytolytic effector cells) or aim at mobilizing endogenous anti-tumor immunity. The latter approach includes dendritic cells (DC) most frequently in the form of in vitro cultured peripheral blood monocytes-derived DC. Human blood γδT cells are selective for a single class of non-peptide agonists ("phosphoantigens") and develop into potent antigen-presenting cells (APC), termed γδT-APC within 1-3 days of in vitro culture. Availability of large numbers of γδT-APC would be advantageous for use as a novel cellular vaccine. We here report optimal γδT cell expansion (>10(7) cells/ml blood) when peripheral blood mononuclear cells (PBMC) from healthy individuals and melanoma patients were stimulated with zoledronate and then cultured for 14 days in the presence of IL-2 and IL-15, yielding γδT cell cultures of variable purity (77 ± 21 and 56 ± 26%, respectively). They resembled effector memory αβT (TEM) cells and retained full functionality as assessed by in vitro tumor cell killing as well as secretion of pro-inflammatory cytokines (IFNγ, TNFα) and cell proliferation in response to stimulation with phosphoantigens. Importantly, day 14 γδT cells expressed numerous APC-related cell surface markers and, in agreement, displayed potent in vitro APC functions. Day 14 γδT cells from PBMC of patients with cancer were equally effective as their counterparts derived from blood of healthy individuals and triggered potent CD8(+) αβT cell responses following processing and cross-presentation of simple (influenza M1) and complex (tuberculin purified protein derivative) protein antigens. Of note, and in clear contrast to peripheral blood γδT cells, the ability of day 14 γδT cells to trigger antigen-specific αβT cell responses did not depend on re-stimulation. We conclude that day 14 γδT cell cultures provide a convenient source of autologous APC for use in immunotherapy of patients with various cancers.
authors.affiliation: Institute of Infection and Immunity, Cardiff University School of Medicine , Cardiff , UK.
authors.forename: Mohd Wajid A
authors.initials: MW
authors.lastname: Khan
authors.affiliation: NIHR Biomedical Research Unit, Centre for Liver Research, University of Birmingham Medical School , Birmingham , UK.
authors.forename: Stuart M
authors.initials: SM
authors.lastname: Curbishley
authors.affiliation: Institute of Infection and Immunity, Cardiff University School of Medicine , Cardiff , UK.
authors.forename: Hung-Chang
authors.initials: HC
authors.lastname: Chen
authors.affiliation: Institute of Infection and Immunity, Cardiff University School of Medicine , Cardiff , UK.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Thomas
authors.affiliation: Department of Immunology, Institute of Medical Microbiology and Hygiene, University of Freiburg , Freiburg , Germany.
authors.forename: Hanspeter
authors.initials: H
authors.lastname: Pircher
authors.affiliation: Unit of Clinical and Experimental Immunology, Humanitas Clinical and Research Center, Rozzano , Milan , Italy ; Department of Medical Biotechnologies and Translational Medicine, University of Milan , Milan , Italy.
authors.forename: Domenico
authors.initials: D
authors.lastname: Mavilio
authors.affiliation: CR-UK Clinical Trials Unit, School of Cancer Sciences, University of Birmingham Medical School , Birmingham , UK.
authors.forename: Neil M
authors.initials: NM
authors.lastname: Steven
authors.affiliation: Institute of Infection and Immunity, Cardiff University School of Medicine , Cardiff , UK.
authors.forename: Matthias
authors.initials: M
authors.lastname: Eberl
authors.affiliation: Institute of Infection and Immunity, Cardiff University School of Medicine , Cardiff , UK.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Moser
keywords: antigen-presentation; vaccine; αβT cells; γδT cells
references.citation: Exp Hematol. 2009 Aug;37(8):956-68
references.pmid: 19409955
references.citation: Br J Haematol. 2009 Jan;144(2):245-50
references.pmid: 19016713
references.citation: Clin Exp Immunol. 2005 Nov;142(2):292-302
references.pmid: 16232216
references.citation: J Leukoc Biol. 2005 Jan;77(1):67-70
references.pmid: 15498816
references.citation: Proc Natl Acad Sci U S A. 2009 Feb 17;106(7):2307-12
references.pmid: 19171897
references.citation: J Exp Med. 1994 Apr 1;179(4):1109-18
references.pmid: 8145033
references.citation: Annu Rev Immunol. 2001;19:225-52
references.pmid: 11244036
references.citation: Br J Cancer. 2011 Sep 6;105(6):778-86
references.pmid: 21847128
references.citation: Clin Cancer Res. 2002 Dec;8(12):3877-84
references.pmid: 12473603
references.citation: Proc Natl Acad Sci U S A. 2005 Oct 11;102(41):14741-6
references.pmid: 16203967
references.citation: Cancer Immunol Immunother. 2008 Nov;57(11):1599-609
references.pmid: 18301889
references.citation: Cancer Immunol Immunother. 2008 Apr;57(4):531-9
references.pmid: 17764010
references.citation: Nature. 1998 Mar 19;392(6673):245-52
references.pmid: 9521319
references.citation: Cytotherapy. 2008;10(8):842-56
references.pmid: 19016372
references.citation: J Immunol. 2007 Apr 1;178(7):4304-14
references.pmid: 17371987
references.citation: J Leukoc Biol. 2006 Apr;79(4):663-6
references.pmid: 16461739
references.citation: Nature. 1999 Oct 14;401(6754):708-12
references.pmid: 10537110
references.citation: Cytotherapy. 2013 Apr;15(4):481-91
references.pmid: 23391461
references.citation: Cancer Immunol Immunother. 2012 Mar;61(3):385-96
references.pmid: 21928126
references.citation: Cancer Immunol Immunother. 2010 Oct;59(10):1521-30
references.pmid: 20563721
references.citation: Anticancer Res. 2011 Jul;31(7):2419-24
references.pmid: 21873153
references.citation: J Immunol Methods. 2007 Sep 30;326(1-2):63-75
references.pmid: 17716681
references.citation: Cancer Immunol Immunother. 2010 Nov;59(11):1611-9
references.pmid: 20582413
references.citation: Nat Rev Cancer. 2005 Apr;5(4):263-74
references.pmid: 15776005
references.citation: Nat Rev Immunol. 2005 Apr;5(4):296-306
references.pmid: 15803149
references.citation: Expert Opin Biol Ther. 2008 Jul;8(7):875-83
references.pmid: 18549319
references.citation: Immunity. 2013 Jul 25;39(1):38-48
references.pmid: 23890062
references.citation: Curr Opin Immunol. 2006 Aug;18(4):512-6
references.pmid: 16777399
references.citation: Anticancer Res. 2010 Feb;30(2):575-9
references.pmid: 20332473
references.citation: Blood. 2000 Jan 1;95(1):39-47
references.pmid: 10607682
references.citation: Science. 2013 Dec 20;342(6165):1432-3
references.pmid: 24357284
references.citation: Blood. 2002 Nov 15;100(10):3698-702
references.pmid: 12393723
references.citation: J Immunol. 2012 Feb 15;188(4):1708-16
references.pmid: 22250090
references.citation: Immunity. 2009 Aug 21;31(2):184-96
references.pmid: 19699170
references.citation: J Immunol. 2002 May 15;168(10):4920-9
references.pmid: 11994442
references.citation: Cancer Immunol Immunother. 2013 Apr;62(4):677-87
references.pmid: 23151944
references.citation: Proc Natl Acad Sci U S A. 2010 May 11;107(19):8730-5
references.pmid: 20413723
references.citation: Cancer Immunol Immunother. 2011 Aug;60(8):1075-84
references.pmid: 21519826
references.citation: J Invest Dermatol. 2013 Jul;133(7):1879-89
references.pmid: 23419694
references.citation: Immunol Lett. 2005 Feb 15;97(1):19-29
references.pmid: 15626472
references.citation: Nat Rev Immunol. 2009 Jul;9(7):480-90
references.pmid: 19543225
references.citation: Eur J Cardiothorac Surg. 2010 May;37(5):1191-7
references.pmid: 20137969
references.citation: Cytotherapy. 2011 Jan;13(1):92-7
references.pmid: 20831354
references.citation: J Immunother. 2009 Apr;32(3):310-21
references.pmid: 19242369
references.citation: J Exp Med. 2003 Aug 4;198(3):391-7
references.pmid: 12900516
references.citation: Trends Pharmacol Sci. 2012 Jan;33(1):35-41
references.pmid: 22032984
references.citation: Eur J Clin Invest. 2011 Nov;41(11):1186-94
references.pmid: 22775565
references.citation: Nat Rev Immunol. 2013 Feb;13(2):88-100
references.pmid: 23348415
references.citation: J Immunol. 2010 Jan 1;184(1):452-65
references.pmid: 19949105
references.citation: Science. 2005 Jul 8;309(5732):264-8
references.pmid: 15933162
references.citation: Nat Immunol. 2013 Sep;14(9):908-16
references.pmid: 23872678
references.citation: Br J Haematol. 2007 Oct;139(2):206-16
references.pmid: 17897296
references.citation: N Engl J Med. 2010 Jul 29;363(5):411-22
references.pmid: 20818862
references.citation: Cancer Res. 2010 Dec 15;70(24):10024-7
references.pmid: 21159627
references.citation: J Immunother. 2011 Mar;34(2):202-11
references.pmid: 21304399
references.citation: Blood. 2012 Sep 13;120(11):2269-79
references.pmid: 22767497
references.citation: Nat Rev Immunol. 2010 Jul;10(7):467-78
references.pmid: 20539306
references.citation: Nature. 2007 Sep 27;449(7161):419-26
references.pmid: 17898760
references.citation: Immunology. 2009 Feb;126(2):256-67
references.pmid: 18775028
references.citation: Eur J Immunol. 2012 Jan;42(1):110-9
references.pmid: 22009762
references.citation: Trends Immunol. 2009 Dec;30(12):562-8
references.pmid: 19853512
references.citation: Blood. 2003 Nov 15;102(10):3693-701
references.pmid: 12881309
references.citation: Immunity. 2013 Jan 24;38(1):13-25
references.pmid: 23352221
references.citation: Cell Mol Life Sci. 2011 Jul;68(14):2443-52
references.pmid: 21573785
references.citation: Cancer Immunol Immunother. 2007 Apr;56(4):469-76
references.pmid: 16850345
references.citation: Cell Mol Immunol. 2012 May;9(3):245-54
references.pmid: 22139198
references.citation: J Immunol. 2009 Nov 1;183(9):5622-9
references.pmid: 19843947
references.citation: J Immunol. 2005 Apr 15;174(8):4606-12
references.pmid: 15814683