Department of Surgery, City of Hope National Medical Center, Duarte, CA 91010, USA.
Cancer Immunotherapeutics Program, Beckman Research Institute of City of Hope National Medical Center, Duarte, CA 91010, USA.
Int J Mol Sci. 2023 Sep 16;24(18):14189. doi: 10.3390/ijms241814189.
We studied the immunotherapeutic potential of CF33-hNIS-antiPDL1 oncolytic virus (OV) against gastric cancer with peritoneal metastasis (GCPM). We collected fresh malignant ascites (MA) or peritoneal washings (PW) during routine paracenteses and diagnostic laparoscopies from GC patients (n = 27). Cells were analyzed for cancer cell markers and T cells, or treated with PBS, CF33-GFP, or CF33-hNIS-antiPDL1 (MOI = 3). We analyzed infectivity, replication, cytotoxicity, CD107α upregulation of CD8 and CD4 T cells, CD274 (PD-L1) blockade of cancer cells by virus-encoded anti-PD-L1 scFv, and the release of growth factors and cytokines. We observed higher CD45/large-size cells and lower CD8 T cell percentages in MA than PW. CD45/large-size cells were morphologically malignant and expressed CD274 (PD-L1), CD252 (OX40L), and EGFR. CD4 and CD8 T cells did not express cell surface exhaustion markers. Virus infection and replication increased cancer cell death at 15 h and 48 h. CF33-hNIS-antiPDL1 treatment produced functional anti-PD-L1 scFv, which blocked surface PD-L1 binding of live cancer cells and increased CD8CD107α and CD4CD107α T cell percentages while decreasing EGF, PDGF, soluble anti-PD-L1, and IL-10. CF33-OVs infect, replicate in, express functional proteins, and kill ex vivo GCPM cells with immune-activating effects. CF33-hNIS-antiPDL1 displays real potential for intraperitoneal GCPM therapy.
我们研究了 CF33-hNIS-antiPDL1 溶瘤病毒 (OV) 在伴有腹膜转移的胃癌 (GCPM) 中的免疫治疗潜力。我们在常规穿刺和诊断性腹腔镜检查期间从 GC 患者 (n = 27) 中收集新鲜的恶性腹水 (MA) 或腹膜灌洗液 (PW)。对细胞进行癌症细胞标志物和 T 细胞分析，或用 PBS、CF33-GFP 或 CF33-hNIS-antiPDL1（MOI = 3）处理。我们分析了感染性、复制、细胞毒性、CD8 和 CD4 T 细胞的 CD107α 上调、病毒编码的抗 PD-L1 scFv 对癌细胞的 CD274（PD-L1）阻断作用，以及生长因子和细胞因子的释放。我们观察到 MA 中的 CD45/大细胞比例高于 PW，而 CD8 T 细胞比例较低。CD45/大细胞形态上为恶性，表达 CD274（PD-L1）、CD252（OX40L）和 EGFR。CD4 和 CD8 T 细胞不表达细胞表面耗竭标志物。病毒感染和复制增加了 15 小时和 48 小时的癌细胞死亡。CF33-hNIS-antiPDL1 治疗产生了功能性的抗 PD-L1 scFv，可阻断活癌细胞表面 PD-L1 的结合，增加 CD8CD107α 和 CD4CD107α T 细胞的比例，同时降低 EGF、PDGF、可溶性抗 PD-L1 和 IL-10。CF33-OV 可感染、复制、表达功能性蛋白，并杀伤体外 GCPM 细胞，具有免疫激活作用。CF33-hNIS-antiPDL1 具有治疗腹膜内 GCPM 的真正潜力。
