研究概要
抗GD2-MSCs使用小鼠IgG在同基因模型中显示出归巢效应和显著的肿瘤生长抑制作用。基于抗GD2-MSC的细胞免疫疗法可能成为难治性神经母细胞瘤的一种新型治疗策略。
研究思路结论见上方概要
目的
开发了分泌小鼠IgG抗双唾液酸神经节苷脂GD2抗体的小鼠间充质干细胞(anti-GD2-MSCs),并在体内神经母细胞瘤小鼠模型中验证了其抗肿瘤效果。
方法
构建了抗GD2抗体构建体,将针对GD2的FLAG标签单链可变区片段与连接序列融合,并将固定部分片段从人IgG改为小鼠IgG和GFP蛋白。该构建体通过慢病毒导入小鼠MSCs。通过皮下移植来自TH-MYCN转基因小鼠的肿瘤组织碎片建立同基因小鼠模型,并通过In vivo imaging system成像验证抗GD2-MSCs的归巢效应。根据局部注射材料将同基因模型分为三组:抗GD2-MSCs联合IL-2、IL-2和PBS。切除肿瘤,并计数自然杀伤(NK)细胞。
结果
抗GD2-MSCs在同基因模型中显示出归巢效应。抗GD2-MSCs联合IL-2显著抑制了皮下肿瘤的生长速率(p < 0.05)。皮下肿瘤免疫染色显示,同一组中NK细胞浸润增加(p < 0.01)。
展开英文摘要原文
PURPOSE: Mouse IgG anti-disialoganglioside GD2 antibody-secreting mouse mesenchymal stem cells (anti-GD2-MSCs) were developed, and their anti-tumor effects were validated in an in vivo neuroblastoma mouse model.
METHODS: Anti-GD2 antibody constructs were generated, incorporating FLAG-tagged single-chain fragment variables against GD2 fused to a linker sequence, and a fragment of a stationary portion was changed from human IgG to mouse IgG and GFP protein. The construct was lentivirally introduced into mouse MSCs. A syngeneic mouse model was established through the subcutaneous transplantation of a tumor tissue fragment from a TH-MYCN transgenic mouse, and the homing effects of anti-GD2-MSCs were validated by In vivo imaging system imaging. The syngeneic model was divided into three groups according to topical injection materials: anti-GD2-MSCs with IL-2, IL-2, and PBS. The tumors were removed, and natural killer (NK) cells were counted.
RESULTS: Anti-GD2-MSCs showed homing effects in syngeneic models. The growth rate of subcutaneous tumors was significantly suppressed by anti-GD2-MSCs with IL-2 (p < 0.05). Subcutaneous tumor immunostaining showed an increased NK cell infiltration in the same group (p < 0.01).
CONCLUSION: Anti-GD2-MSCs using mouse IgG showed a homing effect and significant tumor growth suppression in syngeneic models. Anti-GD2-MSC-based cellular immunotherapy could be a novel therapeutic strategy for intractable neuroblastoma.
论文信息
- 作者
- Kambe K、Iguchi M、Higashi M、Yagyu S、Fumino S、Kishida T、Mazda O、Tajiri T
- 单位
- Department of Pediatric Surgery, Kyoto Prefectural University of Medicine, 465 Kawaramachi-Hirokoji, Kamigyo-Ku, Kyoto, 602-8566, Japan. kosukeka@koto.kpu-m.ac.jp.Japan
- 期刊
- Pediatric surgery international2022 Dec 9