T细胞双特异性抗体（TCB）是癌症免疫治疗领域引人注目的研究重点。最近，肽-MHC（主要组织相容性复合物）靶向TCB作为一种新型生物治疗药物崛起，具有改善的特异性。这些TCB能够同时结合靶细胞由人类白细胞抗原（HLA）等位基因编码的多态性物种特异性MHC所呈现的目标肽和人类T淋巴细胞表达的CD3共受体。然而，传统模型通常对于人体组织上“靶内，非肿瘤”相互作用的评估缺乏预测能力。在这里，我们报道了一种免疫浸润的肾脏器官体模型，其中外周血单个核细胞（PBMCs）以及非靶向（对照）或基于靶向TCB的工具化合物在流动条件下循环。目标由通过双价T细胞受体样结合结构域与HLA-A2共呈现的来自细胞内肿瘤抗原Wilms' tumor 1（WT1）的RMF肽组成。使用我们的模型，在PBMCs和多种工具化合物的存在下，我们测量了TCB介导的RMF-HLA-A2呈现细胞的CD8+ T细胞活化和杀伤。DP47是一种非靶向MHC的TCB，仅与CD3结合（阴性对照），不能促进T细胞活化和杀伤。相反，非特异性的ESK1样TCB（阳性对照）促进CD8+ T细胞扩张，并伴随剂量依赖性T细胞介导的多种细胞杀伤，而WT1-TCB*高特异性识别RMF-HLA-A2复合物，只导致肾脏器官体内WT1表达细胞的选择性杀伤。我们的三维肾脏器官体模型为癌症免疫治疗的临床前测试和研究组织-免疫系统相互作用提供了平台。

T cell bispecific antibodies (TCBs) are the focus of intense development for cancer immunotherapy. Recently, peptide-MHC (major histocompatibility complex)-targeted TCBs have emerged as a new class of biotherapeutics with improved specificity. These TCBs simultaneously bind to target peptides presented by the polymorphic, species-specific MHC encoded by the human leukocyte antigen (HLA) allele present on target cells and to the CD3 coreceptor expressed by human T lymphocytes. Unfortunately, traditional models for assessing their effects on human tissues often lack predictive capability, particularly for "on-target, off-tumor" interactions. Here, we report an immune-infiltrated, kidney organoid-on-chip model in which peripheral blood mononuclear cells (PBMCs) along with nontargeting (control) or targeting TCB-based tool compounds are circulated under flow. The target consists of the RMF peptide derived from the intracellular tumor antigen Wilms' tumor 1 (WT1) presented on HLA-A2 via a bivalent T cell receptor-like binding domain. Using our model, we measured TCB-mediated CD8+ T cell activation and killing of RMF-HLA-A2-presenting cells in the presence of PBMCs and multiple tool compounds. DP47, a non-pMHC-targeting TCB that only binds to CD3 (negative control), does not promote T cell activation and killing. Conversely, the nonspecific ESK1-like TCB (positive control) promotes CD8+ T cell expansion accompanied by dose-dependent T cell-mediated killing of multiple cell types, while WT1-TCB* recognizing the RMF-HLA-A2 complex with high specificity, leads solely to selective killing of WT1-expressing cells within kidney organoids under flow. Our 3D kidney organoid model offers a platform for preclinical testing of cancer immunotherapies and investigating tissue-immune system interactions.