用于模拟免疫疗法肿瘤清除与正常组织免疫损伤的人多器官微生理系统

Despite the clinical success of CTLA-4 blockade, immune checkpoint inhibitors can trigger immune-related adverse events (irAEs) that remain difficult to predict preclinically, in part because conventional in vitro and animal models lack the capacity to simultaneously evaluate ECM-dependent T-cell infiltration, tumor cytotoxicity, and normal tissue immune tolerance. We developed a multiorgan system to quantify how fibrin matrix density, T-cell donor identity, HLA compatibility, and ipilimumab dose collectively shape immune cytotoxic responses across tumor and normal tissues. Imaging experiments compared fibrin concentrations from 2.5-10 mg/mL to assess physical barrier effects on immune infiltration. Brain (SK-N-SH/HMC3), gut (T84) tumor spheroids, or human primary hepatocyte spheroids with or without Kupffer cells were embedded in a defined fibrin hydrogel (6 mg/mL fibrinogen, 4 U/mL thrombin). Tumor models used two T-cell donors, while the liver model was perfused with either matched or mismatched CD8⁺ T-cell donors. Ipilimumab was applied from 0-10 µg/mL. Spheroid density, derived from fluorescence-based viability measurements, was plotted against antibody concentration.T-cell penetration into spheroids was strongly dependent on fibrin density with low-to-moderate matrix (2.5-6 mg/mL) supported deep infiltration, whereas high-density fibrin (8-10 mg/mL) restricted immune access and created an immune-excluded phenotype. CTLA-4 blockade overcame this barrier in part, enabling infiltration even through dense matrices. Dose-response analysis demonstrated robust tumor killing in both brain and gut spheroids, exhibiting a steep viability decline beginning at 1 µg/mL and reached near-complete loss at 10 µg/mL.Response magnitude varied between donors, reproducing clinically observed heterogeneity in immune potency. In contrast, normal hepatocyte spheroids remained intact under matched donor conditions across all doses. However, mismatched T cells produced clear dose-dependent injury that was significantly amplified in the presence of Kupffer cells, revealing a macrophage-dependent mechanism of immune-mediated hepatotoxicity.This multiorgan platform recapitulates fibrin-driven immune exclusion, checkpoint-enhanced infiltration, effective tumor cytotoxicity, and HLA mismatch-dependent normal tissue injury, mirroring key therapeutic and toxic outcomes observed in patients. We evaluated antigen-specific T cells, bispecific antibodies, and antibody-drug conjugates to assess on-target efficacy, off-target toxicity, and bystander effects. These findings support the use of human microphysiological systems as predictive tools for immunotherapy, enabling mechanistic dissection of efficacy-toxicity trade-offs prior to clinical testing and supporting FDA-aligned strategies to reduce non-human primate use in monoclonal antibody safety testing.