title: CTLA-4⁺ Regulatory T Cells Increased in Cetuximab-Treated Head and Neck Cancer Patients Suppress NK Cell Cytotoxicity and Correlate with Poor Prognosis.
abstract: The EGFR-targeted antibody cetuximab is effective against head and neck cancer (HNSCC), but in only 15% to 20% of patients, and the variability and extent of cetuximab-mediated cellular immunity is not fully understood. We hypothesized that regulatory T cells (Treg) may exert a functional and clinical impact on antitumor immunity in cetuximab-treated individuals. The frequency, immunosuppressive phenotype, and activation status of Treg and natural killer (NK) cells were analyzed in the circulation and tumor microenvironment of cetuximab-treated patients with HNSCC enrolled in a novel neoadjuvant, single-agent cetuximab clinical trial. Notably, cetuximab treatment increased the frequency of CD4(+)FOXP3(+) intratumoral Treg expressing CTLA-4, CD39, and TGFβ. These Treg suppressed cetuximab-mediated antibody-dependent cellular cytotoxicity (ADCC) and their presence correlated with poor clinical outcome in two prospective clinical trial cohorts. Cetuximab expanded CTLA-4(+)FOXP3(+) Treg in vitro, in part, by inducing dendritic cell maturation, in combination with TGFβ and T-cell receptor triggering. Importantly, cetuximab-activated NK cells selectively eliminated intratumoral Treg but preserved effector T cells. In ex vivo assays, ipilimumab targeted CTLA-4(+) Treg and restored cytolytic functions of NK cells mediating ADCC. Taken together, our results argue that differences in Treg-mediated suppression contribute to the clinical response to cetuximab treatment, suggesting its improvement by adding ipilimumab or other strategies of Treg ablation to promote antitumor immunity.
authors.affiliation: Department of Otolaryngology, University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Hyun-Bae
authors.initials: HB
authors.lastname: Jie
authors.affiliation: Department of Otolaryngology, University of Pittsburgh, Pittsburgh, Pennsylvania. University Duisburg-Essen, Department of Otorhinolaryngology, Essen, Germany.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Schuler
authors.affiliation: Department of Otolaryngology, University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Steve C
authors.initials: SC
authors.lastname: Lee
authors.affiliation: Department of Otolaryngology, University of Pittsburgh, Pittsburgh, Pennsylvania.
authors.forename: Raghvendra M
authors.initials: RM
authors.lastname: Srivastava
authors.affiliation: Division of Hematology/Oncology, Department of Medicine, UT Health Science Center at San Antonio, Cancer Therapy and Research Center, San Antonio, Texas.
authors.forename: Athanassios
authors.initials: A
authors.lastname: Argiris
authors.affiliation: Department of Surgery, Massachusetts General Hospital, Boston, Massachusetts.
authors.forename: Soldano
authors.initials: S
authors.lastname: Ferrone
authors.affiliation: Department of Otolaryngology, University of Pittsburgh, Pittsburgh, Pennsylvania. Department of Immunology, University of Pittsburgh, Pittsburgh, Pennsylvania. Department of Pathology, University of Pittsburgh, Pittsburgh, Pennsylvania. Cancer Immunology Program, University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania.
authors.forename: Theresa L
authors.initials: TL
authors.lastname: Whiteside
authors.affiliation: Department of Otolaryngology, University of Pittsburgh, Pittsburgh, Pennsylvania. Department of Immunology, University of Pittsburgh, Pittsburgh, Pennsylvania. Cancer Immunology Program, University of Pittsburgh Cancer Institute, Pittsburgh, Pennsylvania. ferrisrl@upmc.edu.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Ferris
mesh_terms: D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage; D015703:Antigens, CD / Q000096:biosynthesis; D001081:Apyrase / Q000096:biosynthesis; D060908:CTLA-4 Antigen / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000068818:Cetuximab; D017322:Clinical Trials, Phase II as Topic; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D005260:Female; D051858:Forkhead Transcription Factors / Q000096:biosynthesis; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006258:Head and Neck Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007694:Killer Cells, Natural / Q000187:drug effects* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D011379:Prognosis; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D016212:Transforming Growth Factor beta / Q000096:biosynthesis
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D015703:Antigens, CD; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D016212:Transforming Growth Factor beta; D001081:Apyrase; C067869:CD39 antigen; D000068818:Cetuximab
references.citation: Dis Esophagus. 2010 Nov;23(8):675-81
references.pmid: 20545975
references.citation: Clin Cancer Res. 2003 Oct 1;9(12):4404-8
references.pmid: 14555512
references.citation: Exp Biol Med (Maywood). 2011 Apr 1;236(4):375-89
references.pmid: 21427235
references.citation: PLoS Pathog. 2011 Jul;7(7):e1002110
references.pmid: 21750674
references.citation: Immunol Res. 2011 Aug;50(2-3):248-54
references.pmid: 21717064
references.citation: J Clin Invest. 2011 Sep;121(9):3609-22
references.pmid: 21841316
references.citation: Breast Cancer Res Treat. 2011 Nov;130(2):465-75
references.pmid: 21308409
references.citation: Cancer Immunol Immunother. 2011 Dec;60(12):1801-10
references.pmid: 21811786
references.citation: Nat Rev Immunol. 2011 Dec;11(12):852-63
references.pmid: 22116087
references.citation: Clin Cancer Res. 2012 Jan 15;18(2):432-41
references.pmid: 22090361
references.citation: Nat Med. 2012 Feb;18(2):199-200
references.pmid: 22310681
references.citation: Cancer Res. 2012 Mar 15;72(6):1407-15
references.pmid: 22258454
references.citation: J Immunother. 2012 Jun;35(5):440-7
references.pmid: 22576349
references.citation: Cancer Res. 2012 Jul 15;72(14):3557-69
references.pmid: 22573714
references.citation: Mol Cancer Ther. 2012 Nov;11(11):2429-39
references.pmid: 22927667
references.citation: Clin Cancer Res. 2013 Apr 1;19(7):1858-72
references.pmid: 23444227
references.citation: Br J Cancer. 2005 Mar 14;92(5):913-20
references.pmid: 15714205
references.citation: J Exp Med. 2005 Oct 17;202(8):1075-85
references.pmid: 16230475
references.citation: J Immunol. 2006 Feb 1;176(3):1582-7
references.pmid: 16424187
references.citation: N Engl J Med. 2006 Feb 9;354(6):567-78
references.pmid: 16467544
references.citation: J Immunol. 2006 Aug 15;177(4):2331-9
references.pmid: 16887994
references.citation: Clin Cancer Res. 2006 Sep 15;12(18):5423-34
references.pmid: 17000676
references.citation: Clin Cancer Res. 2007 Mar 1;13(5):1552-61
references.pmid: 17332301
references.citation: Proc Natl Acad Sci U S A. 2007 Feb 27;104(9):3378-83
references.pmid: 17360654
references.citation: Clin Cancer Res. 2007 Aug 1;13(15 Pt 1):4345-54
references.pmid: 17671115
references.citation: J Immunol Methods. 2007 Aug 31;325(1-2):51-66
references.pmid: 17617419
references.citation: J Immunol. 2007 Oct 15;179(8):5191-203
references.pmid: 17911604
references.citation: Arch Otolaryngol Head Neck Surg. 2007 Dec;133(12):1277-81
references.pmid: 18086972
references.citation: J Immunol. 2008 Apr 1;180(7):4679-86
references.pmid: 18354191
references.citation: J Immunol. 2008 May 15;180(10):6566-76
references.pmid: 18453575
references.citation: Oncogene. 2008 Jun 26;27(28):3944-56
references.pmid: 18297114
references.citation: J Immunol. 2008 Sep 15;181(6):3784-92
references.pmid: 18768831
references.citation: Science. 2008 Oct 10;322(5899):271-5
references.pmid: 18845758
references.citation: J Clin Oncol. 2009 Apr 10;27(11):1864-71
references.pmid: 19289630
references.citation: Cancer Immunol Immunother. 2009 Nov;58(11):1853-64
references.pmid: 19319529
references.citation: J Clin Oncol. 2010 Mar 1;28(7):1254-61
references.pmid: 20100961
references.citation: J Immunol. 2010 Jun 15;184(12):6695-708
references.pmid: 20483724
references.citation: Mol Cancer. 2010;9:139
references.pmid: 20529262
references.citation: Proc Natl Acad Sci U S A. 2009 Dec 8;106(49):20847-52
references.pmid: 19934056
references.citation: J Clin Oncol. 2010 Oct 1;28(28):4390-9
references.pmid: 20697078
references.citation: Oncogene. 2013 Apr 4;32(14):1743-51
references.pmid: 22751118
references.citation: J Exp Med. 2013 Aug 26;210(9):1695-710
references.pmid: 23897981
references.citation: J Exp Med. 2013 Aug 26;210(9):1685-93
references.pmid: 23897982
references.citation: Clin Cancer Res. 2013 Dec 1;19(23):6585-96
references.pmid: 24097865
references.citation: Lung Cancer. 2014 Jul;85(1):81-7
references.pmid: 24780112
references.citation: Br J Cancer. 2013 Nov 12;109(10):2629-35
references.pmid: 24169351
references.citation: J Clin Oncol. 2010 Dec 20;28(36):5294-300
references.pmid: 21079141