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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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 2

TANs acquire an activated phenotype and novel repertoire of chemokine receptors.

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TANs acquire an activated phenotype and novel repertoire of chemokine re...
(A) Expression of the activation markers CD62L and CD54 on CD15hiCD66b+ PBNs. PBNs were isolated from lung cancer patients using anti-CD15 beads. Results represent 1 of 5 experiments. (B) Digestion protocol did not elicit premature activation of resting PBNs. Results represent 1 of 5 experiments. (C) PBNs acquire an activated CD62LloCD54+ phenotype after treatment with TCM in plates with ultralow attachment surface. Each experiment was repeated at least 5 times. (D) A single-cell suspension was obtained from freshly harvested tumor tissues. TANs were gated on CD11b+CD15hiCD66b+ cells and further analyzed for the expression of activation markers. TANs displayed an activated CD62LloCD54+ phenotype. Results represent 1 of 12 experiments. (E) Expression of the activation markers on gated CD11b+CD15hi TANs, distant lung neutrophils (Distant N), and PBNs. (F) Expression of CCR5, CCR7, CXCR3, and CXCR4 was analyzed on gated CD11b+CD15hiCD66b+ TANs, distant lung neutrophils, and PBNs of cancer patients. Bottom: Representative dot plots. Numbers represent the percentage of cells in each quadrant. Top: Summary of all patient data. Error bars represent mean ± SEM. Statistical analyses were performed with repeated-measures 1-way ANOVA with Tukey’s multiple comparison test for CD62L, CD54, CXCR2, CXCR1, and CCR5, and Kruskal-Wallis and Dunn’s multiple comparison tests for CCR7, CXCR3, and CXCR4 (*P ≤ 0.001, **P ≤ 0.01, ***P ≤ 0.05).