Human double negative T cells target lung cancer via ligand-dependent mechanisms that can be enhanced by IL-15

Abstract

BackgroundThe advents of novel immunotherapies have revolutionized the treatment of cancer. Adoptive cellular therapies using chimeric antigen receptor T (CAR-T) cells have achieved remarkable clinical responses in B cell leukemia and lymphoma but the effect on solid tumors including lung cancer is limited. Here we present data on the therapeutic potential of allogeneic CD3+CD4−CD8− double negative T (DNT) cells as a new cellular therapy for the treatment of lung cancer and underlying mechanisms.MethodsDNTs were enriched and expanded ex vivo from healthy donors and phenotyped by flow cytometry. Functionally, their cytotoxicity was determined against primary and established non-small-cell lung cancer (NSCLC) cell lines in vitro or through in vivo adoptive transfer into xenograft models. Mechanistic analysis was performed using blocking antibodies against various cell surface and soluble markers. Furthermore, the role of IL-15 on DNT function was determined.ResultsWe demonstrated that ex vivo expanded DNTs can effectively lyse various human NSCLC cells in vitro and inhibit tumor growth in xenograft models. Expanded DNTs have a cytotoxic phenotype, as they express NKp30, NKG2D, DNAM-1, membrane TRAIL (mTRAIL), perforin and granzyme B, and secrete IFNγ and soluble TRAIL (sTRAIL). DNT-mediated cytotoxicity was dependent on a combination of tumor-expressed ligands for NKG2D, DNAM-1, NKp30 and/or receptors for TRAIL, which differ among different NSCLC cell lines. Furthermore, stimulation of DNTs with IL-15 increased expression of effector molecules on DNTs, their TRAIL production and cytotoxicity against NSCLC in vitro and in vivo.ConclusionHealthy donor-derived DNTs can target NSCLC in vitro and in vivo. DNTs recognize tumors via innate receptors which can be up-regulated by IL-15. DNTs have the potential to be used as a novel adoptive cell therapy for lung cancer either alone or in combination with IL-15.Electronic supplementary materialThe online version of this article (10.1186/s40425-019-0507-2) contains supplementary material, which is available to authorized users.

Developing Allogeneic Double-Negative T Cells as a Novel Off-the-Shelf Adoptive Cellular Therapy for Cancer

“…In addition to cancer types presented in this study, Xu and colleagues showed that DNTs could target pancreatic carcinoma in a xenograft model (35), and we observed significantly inhibited non-small cell lung cancer progression after DNT treatment in xenograft models (manuscript submitted, ref. 36). Notably, DNTs from a single donor could kill cancer cells of different origins (Fig.…”

DNT Cell-based Immunotherapy: Progress and Applications

“…Lee J et al [36] showed that the inhibitory effects of DNTs on leukemia cell toxicity were largely induced by perforin/GZMB. Yao J et al [68] also showed that perforin/GZMB was involved in the process of killing NSCLCs by allogeneic DNTs.…”

“…Chen B et al [67] demonstrated the feasibility and benefits of using DNTs after conventional chemotherapy. Yao J et al [68] recently reported that IL-15 enhanced the killing effect of DNTs. Fang et al [69] showed that DNT tumor infiltration increased when combined with anti-PD-1 treatment, which enhanced DNT cell-mediated antitumor effects.…”

“…Dokouhaki et al [64] found that NKG2D receptor activation promoted soluble TRAIL production of γδ+DNTs, and soluble TRAIL specifically combined with TRAIL receptors expressed on lung cancer cells to induce apoptosis [76]. Yao J et al [68] showed that DNTs expressed twice as much membrane TRAIL after expanding, and the medium supernatant also detected soluble TRAIL, DNTs induced cytotoxicity in non-small cell lung cancer cells (NSCLCs) depending on the TRAIL pathway. Studies have also shown that the concentration of soluble TRAIL is related to its antitumor activity and clinical response [77].…”

“…NKG2D mediates cytotoxicity by combining MHC class I polypeptide-related sequence A and B with UL16-binding protein molecules, which are highly expressed on tumor cells [62,78]. DNAM-1 induces killing by recognizing CD112 and CD155 ligands, which are highly expressed on tumor cells but negligibly expressed on their normal counterparts [68,[79][80][81][82][83][84][85][86]. Lee J et al [36] demonstrated that DNTs recognize and kill acute myeloid cells mainly by activating NKG2D and DNAM-1 receptors.…”