Keyel Michelle E, Furr Kathryn L, Kang Min H, Reynolds C Patrick
Department of Pediatrics, School of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
Departments of Pediatrics and Cell Biology and Biochemistry, School of Medicine, Texas Tech University Health Sciences Center, Lubbock, TX 79430, USA.
J Clin Med. 2023 Sep 27;12(19):6223. doi: 10.3390/jcm12196223.
GD2, a disialoganglioside, is present on the surface of most neuroblastomas, as well as on some other cancers, such as melanoma and osteogenic sarcoma. The anti-GD2 antibody ch14.18 (dinutuximab) has an FDA-registered indication for use as maintenance therapy for high-risk neuroblastoma with cytokines and 13-cis-retinoic acid after myeloablative therapy. Recent studies using immunohistochemistry of tumor or tumor cells in marrow have shown that some neuroblastomas are negative for GD2. Dinutuximab and other anti-GD2 antibodies are increasingly used in combination with cytotoxic chemotherapy for treating relapsed neuroblastoma, so it is important to be able to identify patients with tumor cells with low GD2 expression, as such patients may experience toxicity but not benefit from the antibody therapy. As the most common clinical samples available for relapsed neuroblastoma are bone marrow aspirates, we developed a method to quantify dinutuximab binding density and the frequency of neuroblastoma cells positive for the antibody in bone marrow aspirates. Here, we describe a multi-color flow cytometry assay that employs non-GD2 antibodies to identify neuroblastoma cells in a mixed population (tumor, bone marrow, or blood) and an anti-GD2 antibody to quantify both the frequency and density of GD2 expression on neuroblastoma cells.
GD2是一种双唾液酸神经节苷脂，存在于大多数神经母细胞瘤以及其他一些癌症（如黑色素瘤和成骨肉瘤）的表面。抗GD2抗体ch14.18（地努图希单抗）已获得美国食品药品监督管理局（FDA）注册，可在清髓性治疗后与细胞因子和13-顺式维甲酸联合用作高危神经母细胞瘤的维持治疗。最近利用肿瘤或骨髓中肿瘤细胞的免疫组织化学研究表明，一些神经母细胞瘤GD2呈阴性。地努图希单抗和其他抗GD2抗体越来越多地与细胞毒性化疗联合用于治疗复发性神经母细胞瘤，因此能够识别GD2表达低的肿瘤细胞患者非常重要，因为这类患者可能会出现毒性反应，但无法从抗体治疗中获益。由于复发性神经母细胞瘤最常见的临床样本是骨髓抽吸物，我们开发了一种方法来量化地努图希单抗结合密度以及骨髓抽吸物中对该抗体呈阳性的神经母细胞瘤细胞的频率。在此，我们描述一种多色流式细胞术检测方法，该方法使用非GD2抗体来识别混合群体（肿瘤、骨髓或血液）中的神经母细胞瘤细胞，并使用抗GD2抗体来量化神经母细胞瘤细胞上GD2表达的频率和密度。
