Tumor-associated neutrophils stimulate T cell responses in early-stage human lung cancer
Evgeniy B. Eruslanov, … , Steven M. Albelda, Sunil Singhal
Evgeniy B. Eruslanov, … , Steven M. Albelda, Sunil Singhal
Published November 10, 2014
Citation Information: J Clin Invest. 2014;124(12):5466-5480. https://doi.org/10.1172/JCI77053.
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Research Article Oncology

Tumor-associated neutrophils stimulate T cell responses in early-stage human lung cancer

Abstract

Infiltrating inflammatory cells are highly prevalent within the tumor microenvironment and mediate many processes associated with tumor progression; however, the contribution of specific populations remains unclear. For example, the nature and function of tumor-associated neutrophils (TANs) in the cancer microenvironment is largely unknown. The goal of this study was to provide a phenotypic and functional characterization of TANs in surgically resected lung cancer patients. We found that TANs constituted 5%–25% of cells isolated from the digested human lung tumors. Compared with blood neutrophils, TANs displayed an activated phenotype (CD62LloCD54hi) with a distinct repertoire of chemokine receptors that included CCR5, CCR7, CXCR3, and CXCR4. TANs produced substantial quantities of the proinflammatory factors MCP-1, IL-8, MIP-1α, and IL-6, as well as the antiinflammatory IL-1R antagonist. Functionally, both TANs and neutrophils isolated from distant nonmalignant lung tissue were able to stimulate T cell proliferation and IFN-γ release. Cross-talk between TANs and activated T cells led to substantial upregulation of CD54, CD86, OX40L, and 4-1BBL costimulatory molecules on the neutrophil surface, which bolstered T cell proliferation in a positive-feedback loop. Together our results demonstrate that in the earliest stages of lung cancer, TANs are not immunosuppressive, but rather stimulate T cell responses.

Authors

Evgeniy B. Eruslanov, Pratik S. Bhojnagarwala, Jon G. Quatromoni, Tom Li Stephen, Anjana Ranganathan, Charuhas Deshpande, Tatiana Akimova, Anil Vachani, Leslie Litzky, Wayne W. Hancock, José R. Conejo-Garcia, Michael Feldman, Steven M. Albelda, Sunil Singhal

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Figure 1

Neutrophils infiltrate NSCLC tissue.

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Neutrophils infiltrate NSCLC tissue. (A–C) Lung cancer tissue sections w...
(A–C) Lung cancer tissue sections were stained using 2-color immunohistochemistry for MPO, HLA-DR, and CD3 to visualize neutrophils, APCs, and T lymphocytes, respectively. Representative images are shown. Original magnification, ×10 (A and C, top), ×20 (B, top), ×40 (bottom). (D) Representative dot plots of total tumor cells that define the phenotype of TANs in NSCLC. TANs were defined as CD15hiCD66b+CD11b+CD16intArg1+MPO+IL-5Ra–. Results represent 1 of 20 experiments. Numbers represent the percentage of TANs. (E) Comparative immunohistochemical analysis of TAN density (cells/mm2) in AC (n = 45) and SCC (n = 25) performed by counting of MPO+ cells in the tumor stroma and the tumor islets. Statistical analyses were performed with Student’s t test for unpaired data. (F) The frequency of TANs in AC (n = 31) and SCC (n = 11) determined by flow cytometry as the percentage of CD11b+CD15hiCD66b+ cells among all cells in tumor. Cumulative results from 42 independent experiments are shown in the scatter plot. Student’s t test for unpaired data. (G) The ratio of TANs to other CD15–CD11b+ cells in AC (n = 22) and SCC (n = 9). Mann-Whitney test for unpaired data. For all scatter plots, error bars represent mean ± SEM. (H) PBNs were analyzed for migration in the Neuro Probe ChemoTx system. Each experiment was run in triplicate and repeated at least 3 times. Results of 1 representative experiment are shown. Error bars represent mean ± SEM. Statistical analysis was performed with Kruskal-Wallis and Dunn’s multiple comparison tests (*P ≤ 0.05, **P ≤ 0.01). fMLP, N-formyl-methionyl-leucyl-phenylalanine.