NKT cells are a specialized subset of lipid-reactive T lymphocytes that play direct and indirect roles in immunosurveillance and anti-tumor immunity. Preclinical studies have shown that NKT cell activation via delivery of exogenous glycolipids elicits a significant anti-tumor immune response. Furthermore, infiltration of NKT cells is associated with a good prognosis in several cancers. In this review, we aim to summarize the role of NKT cells in cancer as well as the current strategies and status of NKT cell immunotherapy. This review also examines challenges and future directions for improving the therapy.

Three-dimensional cell culturing to capture a life-like experimental environment has become a versatile tool for basic and clinical research. Mucosal and skin tissues can be grown as “organoids” in a petri dish and serve a wide variety of research questions. Here, we report our experience with human cervical organoids which could also include an immune component, e.g., Langerhans cells. We employ commercially available human cervical keratinocytes and fibroblasts as well as a myeloid cell line matured and purified into langerin-positive Langerhans cells. These are then seeded on a layer of keratinocytes with underlying dermal equivalent. Using about 10-fold more than the reported number in healthy cervical tissue (1–3%), we obtain differentiated cervical epithelium after 14 days with ~1% being Langerhans cells. We provide a detailed protocol for interested researchers to apply the described “aseptic” organoid model for all sorts of investigations—with or without Langerhans cells.

Three-dimensional cell culturing to capture a life-like experimental environment has become a versatile tool for basic and clinical research. Mucosal and skin tissues can be grown as "organoids" in a petri dish and serve a wide variety of research questions. Here, we report our experience with human cervical organoids which could also include an immune component, e.g. Langerhans cells.We employ commercially available human cervical keratinocytes and fibroblasts as well as a myeloid cell line matured and purified into langerin-positive Langerhans cells. These are then seeded on a layer of keratinocytes with underlying dermal equivalent. Using about 10-fold more than the reported number in healthy cervical tissue (1-3%), we obtain differentiated cervical epithelium after 14 days with ~ 1% being Langerhans cells. We provide a detailed protocol for interested researchers to apply the described "aseptic" organoid model for all sorts of investigations-with or without Langerhans cells.

Background Non-small cell lung cancer (NSCLC) is a leading cause of cancer deaths, with a 5-year survival rate of 19%. Improvements in treatment are crucial for reducing both morbidity and mortality. In this study, we examined the therapeutic benefit of combining oncolytic vesicular stomatitis virus (VSV) expressing reovirus-derived fusion-associated small transmembrane (FAST) proteins (p14 or p15), PD-1 checkpoint blockade, and natural killer T (NKT) cell immunotherapy in a genetic mouse model of lung adenocarcinoma. Methods Mice containing a tamoxifen-inducible Cre recombinase gene driven by the (Club cell-secretory protein) CCSP promoter, were crossed with mice containing a Lox-Stop-Lox KRAS G12D mutation and a floxed p53 allele. CCSP-KP mice were treated with VSV-GFP, VSV-p14, or VSV-p15 on days 40, 42, and 44, followed on day 45 by treatment with a-galactosylceramide-loaded dendritic cells to activate NKT cells. Anti-PD-1 (ip. 300 mg) was given once a week for a total of 4 doses (days 48, 55, 62, and 69). MTT assays were performed to examine the oncolytic potential of the VSV-FAST constructs (VSV-p14, -p15, -p14delta, -p14endop15, -p10 ARV, -p10 NBV) in comparison to UV-inactivated VSV and VSV-GFP controls in murine (LLC, CMT-167) and human lung cancer cell lines (A549). Results MTT cell viability assays demonstrate that VSV-FAST constructs VSV-p14, -p15, and chimeric -p14endop15 have increased tumor cell killing ability when compared to VSV-GFP in both mouse and human lung cancer cells. CCSP-KP mice receiving combinatorial treatment of VSV-p14 or VSV-p15, NKT cell activation, and PD-1 blockade exhibited increased overall survival in comparison with untreated or VSV-GFP combination therapy treated mice CCSP-KP mice. Furthermore, signs of morbidity (heavy or labored respirations, hunched posture, weight loss, etc.) were considerably delayed in treated mice. Conclusions Our study demonstrates that VSV-FAST constructs are effective at killing lung cancer cells and when combined with NKT cell immunotherapy and immune checkpoint blockade, can prolong survival in a mouse model of NSCLC. Acknowledgements BJ is funded by a CIHR operating grant (PJT-153285), JDL is funded by CIBC via the BHCRI CRTP 2020-21 Award, a DMRF 2020-22 I3V Graduate Studentship, and a BHCRI DMRF 2022-24 Rosetti Scholarship. Ethics Approval This study was approved by the Dalhousie University Committee on Laboratory Animals; approval number 20-100.