The Gladstone Institutes, San Francisco, CA, USA.
Biomedical Sciences Graduate Program, University of California San Francisco, San Francisco, CA, USA.
Open Biol. 2022 Mar;12(3):210320. doi: 10.1098/rsob.210320. Epub 2022 Mar 2.
Hepatitis C virus (HCV) remains a global public health challenge with an estimated 71 million people chronically infected, with surges in new cases and no effective vaccine. New methods are needed to study the human immune response to HCV since animal models are limited and cancer cell models often show dysregulated immune and proliferative responses. Here, we developed a CD8 T cell and adult stem cell liver organoid system using a microfluidic chip to coculture 3D human liver organoids embedded in extracellular matrix with HLA-matched primary human T cells in suspension. We then employed automated phase contrast and immunofluorescence imaging to monitor T cell invasion and morphological changes in the liver organoids. This microfluidic coculture system supports targeted killing of liver organoids when pulsed with a peptide specific for HCV non-structural protein 3 (NS3) (KLVALGINAV) in the presence of patient-derived CD8 T cells specific for KLVALGINAV. This demonstrates the novel potential of the coculture system to molecularly study adaptive immune responses to HCV in an setting using primary human cells.
丙型肝炎病毒（HCV）仍然是一个全球性的公共卫生挑战，据估计有 7100 万人慢性感染，新发病例激增，而目前尚无有效的疫苗。由于动物模型存在局限性，癌细胞模型通常表现出失调的免疫和增殖反应，因此需要新的方法来研究人类对 HCV 的免疫反应。在这里，我们使用微流控芯片开发了一种 CD8 T 细胞和成人干细胞肝类器官系统，将嵌入细胞外基质的 3D 人肝类器官与悬浮的 HLA 匹配的原代人 T 细胞共培养。然后，我们采用自动化相差和免疫荧光成像来监测 T 细胞在肝类器官中的入侵和形态变化。当用针对 HCV 非结构蛋白 3（NS3）的肽（KLVALGINAV）脉冲刺激时，该微流控共培养系统在存在针对 KLVALGINAV 的患者来源的 CD8 T 细胞的情况下，支持对肝类器官的靶向杀伤。这表明该共培养系统具有新颖的潜力，可使用原代人细胞在 环境中对 HCV 的适应性免疫反应进行分子研究。
