Polyfunctional tumor-reactive T cells are effectively expanded from non-small cell lung cancers, and correlate with an immune-engaged T cell profile

ABSTRACT

Non-small cell lung cancer (NSCLC) is the second most prevalent type of cancer. With the current treatment regimens, the mortality rate remains high. Therefore, better therapeutic approaches are necessary. NSCLCs generally possess many genetic mutations and are well infiltrated by T cells (TIL), making TIL therapy an attractive option. Here we show that T cells from treatment naive, stage I-IVa NSCLC tumors can effectively be isolated and expanded, with similar efficiency as from normal lung tissue. Importantly, 76% (13/17) of tested TIL products isolated from NSCLC lesions exhibited clear reactivity against primary tumor digests, with 0.5%-30% of T cells producing the inflammatory cytokine Interferon (IFN)-γ. Both CD4⁺ and CD8⁺ T cells displayed tumor reactivity. The cytokine production correlated well with CD137 and CD40L expression. Furthermore, almost half (7/17) of the TIL products contained polyfunctional T cells that produced Tumor Necrosis Factor (TNF)-α and/or IL-2 in addition to IFN-γ, a hallmark of effective immune responses. Tumor-reactivity in the TIL products correlated with high percentages of CD103⁺CD69⁺CD8⁺ T cell infiltrates in the tumor lesions, with PD-1^hiCD4⁺ T cells, and with FoxP3⁺CD25⁺CD4⁺ regulatory T cell infiltrates, suggesting that the composition of T cell infiltrates may predict the level of tumor reactivity. In conclusion, the effective generation of tumor-reactive and polyfunctional TIL products implies that TIL therapy will be a successful treatment regimen for NSCLC patients.

KEYWORDS:

• Non-small cell lung cancer
• tumor infiltrating T cells
• tumor reactivity
• cytokines
• polyfunctional T cells

Authors’ contributions

RdG, MMvL, MMvdH, JdJ, PB, CEvdS, RMS, DA, JBAGH, KM, KJH and MCW designed the study; RdG, MMvL, AG, BPN, JJFvH, KM, and OJHMV performed experiments and analysed data, KJH coordinated sample collection and performed pulmonary surgery, JdJ, KM collected and analysed samples. RdG and MCW wrote the manuscript. MCW supervised the study. All authors read and approved the final manuscript.

Acknowledgments

We thank the Flow cytometry facility from Sanquin Research, and the medical assistance staff from the NCI-AvL for technical help.

Availability of data and material

All data generated or analysed during this study are included in this published article [and its supplementary information files].

Competing interests

The authors declare no competing interests

Disclosure of Potential Conflicts of Interest

No potential conflicts of interest were disclosed.

Ethics approval and consent to participate

The study was performed according to the Declaration of Helsinki (seventh revision, 2013), and executed with consent of the Institutional Review Board of the Netherlands Cancer Institute-Antoni van Leeuwenhoek Hospital (NKI-AvL), Amsterdam, the Netherlands (CFMPB317).

Supplemental material

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Additional information

Funding

This research was supported by intramural funding of Sanquin (PPOC-14-46); Stichting Sanquin Bloedvoorziening [PPOC 14-46].