医学
免疫系统
癌症
免疫疗法
癌症影像学
免疫学
肿瘤科
癌症研究
内科学
作者
Francesca Chiovaro,Irina Agarkova,P. Vonschallen,Silvan Strebel,Armin Wolf
标识
DOI:10.1016/j.annonc.2021.08.1384
摘要
Dissecting the interaction of tumor cells with immune cells is essential to decipher the mechanism of immunotherapies. With our immune-competent 3D InSightTM tumor models we propose optimized in vivo-like conditions over standard cell cultures to assess how immune cells elicit effective responses against cancer. Moreover, we present a live-cell imaging approach to assess the efficacy of immune-targeting therapies with high-content imaging and analysis (HCA) techniques. The lung carcinoma 3D in vitro model was generated using GFP-A549 cells in coculture with dermal fibroblasts and PBMCs in Akura™ plates. PBMCs were previously stimulated either with cytokines or anti-CD3/CD28 antibodies. Using the Yokogawa CQ1 HCA instrument, we monitored the immune cell attack on tumors in real-time and in 3 dimensions without any discernible phototoxicity. Tumor killing activity and T-cell effector function were evaluated by measuring tumor size by automatic stage fluorescence microscopy (Leica Dmi8) or confocal HCA (Yokogawa CQ1). Release of inflammatory cytokines IL-6, TNF, IFNγ and GM-CSF was measured by a multiplex assay (MAGPIX™ Luminex system). Finally, we calculated tumor and immune cell volumes from individual image stacks to evaluate tumor killing and proliferation/infiltration of immune cells. Furthermore, beside modulating the adaptive immune system through T-cell activation, we also integrated our immune-competent 3D tumors with myeloid cells to recapitulate the role of the innate immune system. Our real time confocal analysis of tumor-immune cell cocultures shows that activation of PBMCs with cytokines or CD3/CD28 antibodies drives T-cell-mediated tumor cell killing. Our results demonstrate how this assay platform may be used for I-O studies with engineered T-cells (e.g., CAR-T) or immunomodulatory agents, e.g. bispecific antibodies and immune checkpoint inhibitors. Thanks to the integration of innate immune cells into our immune-competent 3D tumor models, we can also employ this system to reprogram immunosuppressive myeloid cells and unravel mechanisms of innate resistance to cancer immunotherapies.
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