TY - JOUR
T1 - Mechanisms driving neutrophil-induced t-cell immunoparalysis in ovarian cancer
AU - Emmons, Tiffany R.
AU - Giridharan, Thejaswini
AU - Singel, Kelly L.
AU - Khan, A. N. M. Nazmul H.
AU - Ricciuti, Jason
AU - Howard, Kaitlyn
AU - Silva-del Toro, Stephanie L.
AU - Debreceni, Ivy L.
AU - Aarts, Cathelijn E. M.
AU - Brouwer, Mieke C.
AU - Suzuki, Sora
AU - Kuijpers, Taco W.
AU - Jongerius, Ilse
AU - Allen, Lee-Ann H.
AU - Ferreira, Viviana P.
AU - Schubart, Anna
AU - Sellner, Holger
AU - Eder, J. rg
AU - Holland, Steven M.
AU - Ram, Sanjay
AU - Lederer, James A.
AU - Eng, Kevin H.
AU - Moysich, Kirsten B.
AU - Odunsi, Kunle
AU - Yaffe, Michael B.
AU - Zsiros, Emese
AU - Segal, Brahm H.
N1 - Funding Information: This work was supported by Roswell Park Cancer Center Support Grant P30CA016056, 5R01CA188900 (B.H. Segal and K.B. Moysich), National Institute of Allergy and Infectious Diseases (NIAID) R01 AI119965 (L.-A.H. Allen), a National Science Foundation predoctoral fellowship (S.L. Silva-Del Toro), U.S. Department of Defense W81XWH-16-1-0464 (J.A. Lederer), NIH NIAID U01AI138318-03 (J.A. Lederer), and a grant from the Charles and Marjorie Holloway Foundation (M.B. Yaffe). Cytometry services were provided by the Flow and Image Cytometry Shared Resource at the Roswell Park Comprehensive Cancer Center, which is supported in part by the NCI Cancer Center Support Grant 5P30 CA016056. The ImagestreamX-MKII instrument was funded through NIH Shared Instrument Grant 1S10OD018048. This work used resources and services provided by the Translational Imaging Shared Resource at Roswell Park that are supported by NCI P30CA16056. This study used the Clinical Research Services, the University at Buffalo Confocal Microscope and Flow Cytometry Facility, the Harvard Medical Area CyTOF core, and the University of Iowa Central Microscopy Research Facility. The authors thank Dr. Jesus G. Valenzuela (NIH NIAID) for the donation of SALO. Publisher Copyright: © 2021 American Association for Cancer Research.
PY - 2021/7/1
Y1 - 2021/7/1
N2 - T-cell activation and expansion in the tumor microenvironment (TME) are critical for antitumor immunity. Neutrophils in theTME acquire a complement-dependent T-cell suppressor phenotype that is characterized by inhibition of T-cell proliferation and activation through mechanisms distinct from those of myeloid-derived suppressor cells. In this study, we used ascites fluid supernatants (ASC) from patients with ovarian cancer as an authentic component of the TME to evaluate the effects of ASC on neutrophil function and mechanisms for neutrophil-driven immune suppression. ASC prolonged neutrophil life span, decreased neutrophil density, and induced nuclear hypersegmentation. Mass cytometry analysis showed that ASC induced 15 distinct neutrophil clusters. ASC stimulated complement deposition and signaling in neutrophils, resulting in surface mobilization of granule constituents, including NADPH oxidase. NADPH oxidase activation and phosphatidylserine signaling were required for neutrophil suppressor function, although we did not observe a direct role of extracellular reactive oxygen species in inhibiting T-cell proliferation. Postoperative surgical drainage fluid also induced a complement-dependent neutrophil suppressor phenotype, pointing to this effect as a general response to injury. Like circulating lymphocytes, ASC-activated neutrophils caused complement-dependent suppression of tumorassociated lymphocytes. ASC-activated neutrophils adhered to T cells and caused trogocytosis of T-cell membranes. These injury and signaling cues resulted in T-cell immunoparalysis characterized by impaired NFAT translocation, IL2 production, glucose uptake, mitochondrial function, and mTOR activation. Our results demonstrate that complement-dependent priming of neutrophil effector functions in the TME induces a T-cell nonresponsiveness distinct from established checkpoint pathways and identify targets for immunotherapy.
AB - T-cell activation and expansion in the tumor microenvironment (TME) are critical for antitumor immunity. Neutrophils in theTME acquire a complement-dependent T-cell suppressor phenotype that is characterized by inhibition of T-cell proliferation and activation through mechanisms distinct from those of myeloid-derived suppressor cells. In this study, we used ascites fluid supernatants (ASC) from patients with ovarian cancer as an authentic component of the TME to evaluate the effects of ASC on neutrophil function and mechanisms for neutrophil-driven immune suppression. ASC prolonged neutrophil life span, decreased neutrophil density, and induced nuclear hypersegmentation. Mass cytometry analysis showed that ASC induced 15 distinct neutrophil clusters. ASC stimulated complement deposition and signaling in neutrophils, resulting in surface mobilization of granule constituents, including NADPH oxidase. NADPH oxidase activation and phosphatidylserine signaling were required for neutrophil suppressor function, although we did not observe a direct role of extracellular reactive oxygen species in inhibiting T-cell proliferation. Postoperative surgical drainage fluid also induced a complement-dependent neutrophil suppressor phenotype, pointing to this effect as a general response to injury. Like circulating lymphocytes, ASC-activated neutrophils caused complement-dependent suppression of tumorassociated lymphocytes. ASC-activated neutrophils adhered to T cells and caused trogocytosis of T-cell membranes. These injury and signaling cues resulted in T-cell immunoparalysis characterized by impaired NFAT translocation, IL2 production, glucose uptake, mitochondrial function, and mTOR activation. Our results demonstrate that complement-dependent priming of neutrophil effector functions in the TME induces a T-cell nonresponsiveness distinct from established checkpoint pathways and identify targets for immunotherapy.
UR - http://www.scopus.com/inward/record.url?scp=85109040656&partnerID=8YFLogxK
U2 - https://doi.org/10.1158/2326-6066.CIR-20-0922
DO - https://doi.org/10.1158/2326-6066.CIR-20-0922
M3 - Article
C2 - 33990375
SN - 2326-6066
VL - 9
SP - 790
EP - 810
JO - Cancer immunology research
JF - Cancer immunology research
IS - 7
ER -