目的：通过向C57Bl/6J 小鼠腹腔注射 IFN- γ 腺病毒（Ad-mIFN- γ）建立细胞因子释放综合征（CRS）的动物模型。方法：构建Ad-mIFN-γ 及对照Ad-lacZ 腺病毒载体，分别以MOI=100 体外转染小鼠腹腔巨噬细胞，流式细胞术检测其对细胞mIFN-γ 分泌的影响。将40 只雌性C57Bl/6J 小鼠按随机数字表法分为对照组、载体对照组、病毒低、中、高剂量组（每组8只），分别向各组小鼠腹腔注射PBS（200 μL）、Ad-lacZ（2×107 PFU/只）、Ad-mIFN- γ（5×106 PFU/只）、Ad-mIFN-γ（1.5×107 PFU/只）和Ad-mIFN-γ（2×107 PFU /只）。每日观测小鼠的体质量及生存情况；第3 天时采用流式细胞术检测小鼠外周血和脾内单核细胞（CD11b+）、巨噬细胞（CD11b+/CD86+）比例，免疫荧光染色法检测脾内CD11b+的单核细胞比例；第9天时采用流式细胞术检测小鼠血清中细胞因子的分泌水平；第14 天，采用颈椎脱臼法处死小鼠，H-E 染色法观察小鼠肝、脾、肺和肾的病理和组织学变化。结果： Ad-mIFN-γ体外感染小鼠腹腔巨噬细胞，在第3天检测到巨噬细胞分泌mIFN-γ达到峰值（118.34±2.90）pg/mL，并在一周内持续高分泌mIFN-γ，Ad-lacZ 对照组IFN-γ分泌水平较低后，第3天时为（0.17±0.08）pg/mL。小鼠腹腔注射Ad-mIFN-γ后，在14 d内病毒低、中剂量组无小鼠死亡，病毒高剂量组小鼠体质量持续减轻（P<0.001）；第3天，病毒高剂量组小鼠外周血和脾组织内单核细胞、巨噬细胞比例较对照组和中剂量组均显著增加（P<0.05 或P<0.01）；第9 天，病毒低、中、高剂量组小鼠血清中mIFN-γ、IL-6、单核细胞趋化蛋白-1（MCP-1）、IL-1、TNF-α 等细胞因子的水平均显著升高（P<0.001）；10 d 内病毒高剂量组小鼠死亡率达100%。组织病理检测可见病毒高剂量组小鼠的肝、脾、肺、肾组织有明显损伤。结论： Ad-mIFN-γ体外感染小鼠原代腹腔巨噬细胞后，可以快速分泌mIFN-γ；腹腔注射高剂量（2×107 PFU/只）Ad-mIFN-γ导致小鼠出现CRS典型表现，可作为CAR-T细胞治疗诱发CRS的动物模型。

Objective: To establish an animal model of cytokine release syndrome (CRS) by intraperitoneal injection of mouse IFN-γ adenovirus (Ad-mIFN- γ) into C57Bl/6J mice. Methods: Ad-mIFN- γ and Ad-LacZ control adenoviral vectors were constructed, and mouse peritoneal macrophages were transfected with MOI=100 in vitro. The effect on mIFN-γ secretion levels of cells were detected by flow cytometry. Forty female C57Bl/6J mice were divided into the control group, the vector control group, and the low-, medium-, and high-dose virus groups (8 mice in each group) according to the random number table method. The mice were intraperitoneally injected with PBS (200 μL/piece), Ad-lacZ (2×107 PFU/piece), Ad-mIFN-γ (5×106 PFU/piece), Ad-mIFN-γ (1.5×107 PFU/piece) and Ad-mIFN-γ (2×107 PFU/piece), respectively. The body weight and survival of the mice were observed daily. On the third day, flow cytometry was used to detect the proportion of monocytes (CD11b+) and macrophages (CD11b+/CD86+) in the peripheral blood and the spleen, and the proportion of CD11b+ monocytes in the spleen was detected by immunofluorescence staining. On the 9th day, flow cytometry was used to detect the secretion of cytokines in the serum of the mice. On the 14th day, the mice were sacrificed by cervical dislocation, and H-E staining was used to observe the pathological and histological changes of the liver, spleen, lungs and kidneys of the mice. Results: Ad-mIFN-γ infected mouse peritoneal macrophages in vitro, and the level of mIFN-γ secreted by macrophages was detected to reach a peak of (118.34±2.90) pg/mL on the third day, and the secretion level of mIFN-γ continued to be high for one week; while the Ad-lacZ control group secreted a lower level of IFN- γ, with a value of (0.17±0.08) pg/mL on the third day. After intraperitoneal injection of Ad-mIFN-γ, no mice died in the low- and medium-dose virus groups within 14 days, and the body weight of the mice in the high-dose virus group continued to decrease (P<0.001). On the third day, the proportions of monocytes and macrophages in the peripheral blood and spleen tissues of the mice in the high-dose virus group were significantly higher than those in the control group and the medium- dose group (P<0.05 or P<0.01). On the 9th day, the serum levels of mIFN-γ, IL-6, monocyte chemotactic protein-1 (MCP-1), IL-1 and TNF-α in the low-, medium- and high-dose groups increased significantly (P<0.001). Within 10 days, the mortality rate of the mice in the high-dose virus group reached 100%. Histopathological examination showed significant damage in the liver, spleen, lung and kidney tissues of the mice in the high-dose virus group. Conclusion: Mouse primary peritoneal macrophages can rapidly secrete mIFN-γ after infection with Ad-mIFN-γ in vitro. Intraperitoneal injection of high-dose Ad-mIFN-γ (2×107 PFU/piece) resulted in typical CRS manifestations in the mice, which can be used as an animal model for CAR-T cell therapy-induced CRS.

国家自然科学基金（No. 82001206）；江苏省中医药科技发展计划（No. ZT202106）；江苏省中医院高峰学术人才培养工程（No. y2021rc42）；江苏省研究生科研与实践创新计划（No. KYCX23_2122）；医学免疫学国家重点实验室开放课题（No.NKMI2021K18）