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-1hiCD4+ 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.

T cells expanded from renal cell carcinoma display tumor-specific CD137 expression but lack significant IFN-γ, TNF-α or IL-2 production

“… 42 , 54 Similarly, 76% of our NSCLC-derived TIL products showed tumor-specific cytokine responses to autologous tumors, of which 25% was even polyfunctional. 21 Thus, the restricted tumor-specific cytokine production we observed for RCC-derived TIL products is tumor-type specific. The relative absence of tumor-specific cytokine production could not be attributed to an overall inability to produce these cytokines, as the RCC TILs produced ample amounts upon stimulation with PMA/ionomycin.…”

“…These percentages of FOXP3 + T cells are substantially lower than those measured in melanoma or ovarian cancer lesions (~15% of CD3 + T cells). 21 …”

Robust Production of Merkel Cell Polyomavirus Oncogene Specific T Cells From Healthy Donors for Adoptive Transfer

“…CD4 + enriched cells grown in the enhanced cytokine conditions also produced a statistically significant increase of IL-17A-producing CD4 + T cells ( Figure 2E ). Additionally, MCPyV TAg-specific cells gated on CD3 + CD4 + TNFα + cells were polyfunctional for IL-2 and the Th1 effector molecules granzyme B, IFNγ, and TNFα ( Figure 2F ), a feature associated with immune-mediated tumor clearance ( 28 , 29 ). Cytotoxicity assays demonstrated that expanded MCPyV TAg-specific T cells do not induce killing of peptide pulsed autologous moDCs or phytohemagglutinin induced blasts (data not shown).…”

Deletion of p53 and Hyper-Activation of PIK3CA in Keratin-15+ Stem Cells Lead to the Development of Spontaneous Squamous Cell Carcinoma

“…The downloaded RNA-seq data from HNSCC tumor biopsies were uploaded onto CIBERSORT (https://cibersort.stanford.edu/index.php). CIBERSORT input the RNA-seq data into a matrix of reference gene expression signatures (LM22), which contains 547 genes that distinguish phenotypes of 22 human hematopoietic cell [31,32]. This output was used to estimate the relative proportions of the immune cell type composition of a tumor biopsy.…”