南方医科大学学报

南方医科大学学报 ›› 2026, Vol. 46 ›› Issue (6): 1331-1338.doi: 10.12122/j.issn.1673-4254.2026.06.013

• • 上一篇    

CD4/TGF-β双特异性抗体在治疗恶性黑色素瘤腹膜转移小鼠模型中的疗效

江千里1,2(), 刘涛菘3(), 王子展3, 江汕3, 吴海扬3, 林永臻4, SHIYING LU5, 陈灵熙6, 张嘉兴4, 唐露霞3, Raut Rakesh kumar5, 许重远1()   

  1. 1.南方医科大学南方医院，药物临床试验中心，广东 广州 510515
    2.南方医科大学南方医院，血液科，广东 广州 510515；南方医科大学，广东 广州 510515
    3.南方医科大学南方医院，第一临床医学院，广东 广州 510515
    4.南方医科大学南方医院，国际教育学院，广东 广州 510515
    5.南方医科大学南方医院，口腔医学院，广东 广州 510515
    4.广州迈世生物科技有限责任公司，广东 广州 510660
  • 收稿日期:2026-01-16 出版日期:2026-06-20 发布日期:2026-06-24
  • 通讯作者: 许重远 E-mail:jiangqianlid@qq.com;troy-ts@foxmail.com;nflcyljd@smu.edu.cn
  • 作者简介:江千里，副主任医师，硕士生导师，E-mail: jiangqianlid@qq.com
    刘涛菘，在读本科生，E-mail: troy-ts@foxmail.com
    第一联系人：同等贡献作者
  • 基金资助:
    广东省自然科学基金(2016A030313585);广东省自然科学基金(2018A030313647);广州市科技计划-重点研发计划(2024B03J0232);南方医院科研发展基金(K51701059);南方医科大学大学生创新创业训练项目(202412121329);南方医科大学国际教育学院教育课题(2024GJG004)

Efficacy of CD4/TGF-β bispecific antibody in a mouse model of peritoneal metastasis of malignant melanoma

Qianli JIANG1,2(), Taosong LIU3(), Zizhan WANG3, Shan JIANG3, Haiyang WU3, Yongzhen LIN4, LU SHIYING5, Lingxi CHEN6, Jiaxing ZHANG4, Luxia TANG3, Rakesh Kumar Raut5, Zhongyuan XU1()   

  1. 1.Clinical Trial Center, ?Nanfang Hospital, ?Southern Medical University, Guangzhou? 510515, China
    2.Department of Hematology, ?Nanfang Hospital, ?Southern Medical University, Guangzhou? 510515, China
    3.First School of Clinical Medicine, Southern Medical University, Guangzhou 510515, China
    4.School of International Education, Southern Medical University, Guangzhou 510515, China
    5.School of Stomatology, Southern Medical University, Guangzhou 510515, China
    4.Guangzhou Maishi Biotechnology Co. Ltd. , Guangzhou 510660, China
  • Received:2026-01-16 Online:2026-06-20 Published:2026-06-24
  • Contact: Zhongyuan XU E-mail:jiangqianlid@qq.com;troy-ts@foxmail.com;nflcyljd@smu.edu.cn

RichHTML

23

PDF

75

摘要：

目的 探讨新药CD4/TGF-β双特异性抗体对腹膜转移的恶性黑色素瘤的疗效。 方法 药物安全性试验：将20只C57BL/6J背景的人CD4转基因小鼠随机分为3个实验组与1个对照组（每组5只），分别给予2.5、5、10 mg/kg CD4/TGF-β双特异性抗体和等量的磷酸缓冲液，观察小鼠状态、体质量和体温变化。肿瘤模型与药物疗效对比：将20只C57BL/6J背景的人CD4转基因小鼠，随机均分为经典腹腔接种治疗（P4T）组、经典腹腔接种对照（P4U）组、磁力诱导细胞（MagIC）法腹膜接种治疗（M4T）组和MagIC法接种对照（M4U）组（每组5只）。每只小鼠于腹腔内接种5×106磁化的、携带绿色荧光蛋白和荧光素酶的小鼠恶性黑色素瘤细胞B16F10-GL，P4T和M4T组造模3 d后给予CD4/TGF-β双特异性抗体300 μg、每周2次治疗，而M4U和P4U组给予等量磷酸缓冲液。观察小鼠存活和一般情况，并在不同时间点，通过小动物活体成像系统监测体内荧光信号分布及强弱；造模14 d处死小鼠，以大体观察、病理切片、免疫荧光染色和RT-qPCR等方法观测肿瘤负荷和转移情况。 结果 药物安全性试验：各实验组小鼠活动度、体质量、体温变化与对照组相比无显著差异，未出现不良反应。M4U组和P4U组成瘤时间短（P=0.003）；M4U组于壁腹膜固定位置精准成瘤，组内模型鼠成瘤一致性强，而P4U组早期即在腹腔内广泛转移；两组存活时间无明显差异，但M4U组治疗窗口期较P4U组延长（P=0.002）。疗效对比：小动物活体成像显示，同期M4T组肿瘤负荷低于M4U组（P<0.0001），且M4T组肿瘤负荷显著低于P4T组（P=0.0239）；但P4T组肿瘤负荷下降与P4U组相比差异无统计学意义。病理和免疫荧光染色显示M4T和P4T治疗组瘤灶均被致密的Ⅲ型胶原包裹，而相应对照组的瘤灶仅有少数Ⅲ型胶原包裹其内部较多的肿瘤细胞；RT-qPCR显示M4T组和P4T组与各自对照组相比肿瘤负荷相关MITF基因表达量下降（P<0.05），且M4T组下降更明显（P<0.05）。 结论 CD4/TGF-β双特异性抗体对黑色素瘤腹膜转移模型有显著疗效，为其进一步临床应用打下基础。

关键词: 腹膜转移, 黑色素瘤, 双特异性抗体, 免疫治疗, 动物肿瘤模型

Abstract:

Objective To evaluate the efficacy of CD4/TGF-β bispecific antibody in a mouse model of peritoneal metastasis of melanoma. Methods For drug safety testing, 20 human CD4 transgenic C57BL/6J mice were randomized into 4 groups (n=5) for intravenous injections of PBS or 2.5, 5, or 10 mg/kg CD4/TGF‑β bispecific antibody, and the changes in general condition, body weight and body temperature were observed. Another 20 transgenic C57BL/6J mice were randomized into two groups to receive intraperitoneal injection of magnetized B16F10-GL melanoma cells expressing green fluorescent protein and luciferase with or without application of a magnet (3 mm in diameter) to the right abdominal skin before cell injection. Three days later, each group was further divided into two groups for treatment with PBS or CD4/TGF-β bispecific antibody (300 μg) twice a week. In vivo imaging was performed at different time points to assess fluorescence distribution and intensity. On day 14, the mice were euthanized and tumor burden and dissemination were evaluated by gross observation, histopathological analysis, immunofluorescence staining, and RT-qPCR. Results Injection of the antibody did not produce any significant adverse effects in the mice. In the tumor-bearing mice, the application of a magnet significantly accelerated tumor development (3.80±1.79 vs 9.20±2.17 days; P=0.003) and resulted in precise and consistent tumor formation in the parietal peritoneum. Magnet application did not significantly affect survival of the mice but significantly prolonged the therapeutic window (4.60±1.95 vs 10.00±1.73 days; P=0.002). The mice treated with CD4/TGF‑β bispecific antibody had significantly reduced tumor burden irrespective of the inoculation approach, and showed dense encapsulation of the tumor foci by type III collagen, whereas minimal type III collagen deposition and abundant tumor cells were observed in PBS-treated mice. Treatment with the antibody significantly downregulated the expression of melanoma-associated gene MITF, and the reduction was more pronounced in the magnet group. Conclusion The CD4/TGF‑β bispecific antibody shows significant antitumor efficacy and good safety in the mouse model of peritoneal metastasis of melanoma.

Key words: peritoneal metastasis, melanoma, bispecific antibody, immunotherapy, tumor-bearing mouse models