Journal article
Checkpoint blockade accelerates a novel switch from an NKT-driven TNF alpha response toward a T cell driven IFN-gamma response within the tumor microenvironment
Journal for immunotherapy of cancer, Vol.9(6), p.e002269
01/01/2021
DOI: 10.1136/jitc-2020-002269
PMCID: PMC8211075
PMID: 34135102
Abstract
Background
The temporal response to checkpoint blockade (CB) is incompletely understood. Here, we profiled the tumor infiltrating lymphocyte (TIL) landscape in response to combination checkpoint blockade at two distinct timepoints of solid tumor growth.
Methods
C57BL/6 mice bearing subcutaneous MC38 tumors were treated with anti-PD-1 and/or anti-CTLA-4 antibodies. At 11 or 21 days, TIL phenotype and effector function were analyzed in excised tumor digests using high parameter flow cytometry. The contributions of major TIL populations toward overall response were then assessed using ex vivo cytotoxicity and in vivo tumor growth assays.
Results
The distribution and effector function among 37 distinct TIL populations shifted dramatically between early and late MC38 growth. At 11 days, the immune response was dominated by Tumor necrosis factor alpha (TNF alpha)-producing NKT, representing over half of all TIL. These were accompanied by modest frequencies of natural killer (NK), CD4(+), or CD8(+) T cells, producing low levels of IFN-gamma. At 21 days, NKT populations were reduced to a combined 20% of TIL, giving way to increased NK, CD4(+), and CD8(+) T cells, with increased IFN-gamma production. Treatment with CB accelerated this switch. At day 11, CB reduced NKT to less than 20% of all TIL, downregulated TNF alpha across NKT and CD4(+) T cell populations, increased CD4(+) and CD8(+) TIL frequencies, and significantly upregulated IFN-gamma production. Degranulation was largely associated with NK and NKT TIL. Blockade of H-2kb and/or CD1d during ex vivo cytotoxicity assays revealed NKT has limited direct cytotoxicity against parent MC38. However, forced CD1d overexpression in MC38 cells significantly diminished tumor growth, suggesting NKT TIL exerts indirect control over MC38 growth.
Conclusions
Despite an indirect benefit of early NKT activity, CB accelerates a switch from TNF alpha, NKT-driven immune response toward an IFN-gamma driven CD4(+)/CD8(+) T cell response in MC38 tumors. These results uncover a novel NKT/T cell switch that may be a key feature of CB response in CD1d(+) tumors.
Details
- Title: Subtitle
- Checkpoint blockade accelerates a novel switch from an NKT-driven TNF alpha response toward a T cell driven IFN-gamma response within the tumor microenvironment
- Creators
- Shota Aoyama - Moffitt Cancer CenterRyosuke Nakagawa - Department of ImmunologySatoshi Nemoto - Department of ImmunologyPatricio Perez-Villarroel - Moffitt Cancer CenterJames J. Mule - Moffitt Cancer CenterAdam William Mailloux - Roy J. and Lucille A. Carver College of Medicine
- Resource Type
- Journal article
- Publication Details
- Journal for immunotherapy of cancer, Vol.9(6), p.e002269
- DOI
- 10.1136/jitc-2020-002269
- PMID
- 34135102
- PMCID
- PMC8211075
- NLM abbreviation
- J Immunother Cancer
- ISSN
- 2051-1426
- eISSN
- 2051-1426
- Publisher
- Bmj Publishing Group
- Number of pages
- 14
- Grant note
- V Foundation Dr. Miriam and Sheldon G. Adelson Medical Research Foundation P30-CA076292 / Flow Cytometry Shared Resource at the H. Lee Moffitt Cancer Center and Research Institute, a National Cancer Institute (NCI)-designated Comprehensive Cancer Center Cindy and Jon Gruden Fund Chris Sullivan Fund 1R01 CA148995; 1R01 CA184845; P30 CA076292; P50 CA168536; R21 CA214285-01A1 / NCI-NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Cancer Institute (NCI)
- Language
- English
- Date published
- 01/01/2021
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984297319802771
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