Division of Surgical Sciences, Department of Surgery, Duke University Medical Center, Durham, NC, USA.
Department of Neurosurgery, Duke University Medical Center, Durham, NC, USA.
Methods Mol Biol. 2022;2410:609-626. doi: 10.1007/978-1-0716-1884-4_33.
Our group has employed methodologies for effective ex vivo generation of dendritic cell (DC) vaccines for patients with primary malignant brain tumors. In order to reliably produce the most potent, most representational vaccinated DC that will engender an antitumor response requires the ability to orchestrate multiple methodologies that address antigen cross-presentation, T-cell costimulation and polarization, and migratory capacity. In this chapter, we describe a novel method for augmenting the immunogenicity and migratory potential of DCs for their use as vaccines. We have elucidated methodologies to avoid the phenomenon known as immunodominance in generating cancer vaccines. We have found that culturing DC progenitors in serum-free conditions for the duration of the differentiation protocol results in a more homogeneously mature population of DCs that exhibit enhanced immunogenicity compared to DCs generated in serum-containing culture conditions. Furthermore, we demonstrate our method for generating high mobility DCs that readily migrate toward lymphoid organ chemoattractants using CCL3 protein. The combination of these two approaches represents a facile and clinically tractable methodology for generating highly mature DCs with excellent migratory capacity.
我们的团队采用了有效的方法来为原发性脑恶性肿瘤患者体外生成树突状细胞（DC）疫苗。为了可靠地生成最有效、最具代表性的接种 DC，以引发抗肿瘤反应，需要有能力协调多种方法，以解决抗原交叉呈递、T 细胞共刺激和极化以及迁移能力。在本章中，我们描述了一种增强 DC 免疫原性和迁移潜力的新方法，将其用作疫苗。我们已经阐明了避免在生成癌症疫苗中出现免疫优势现象的方法。我们发现，在无血清条件下培养 DC 祖细胞，持续整个分化方案，可产生更均匀成熟的 DC 群体，与在含血清的培养条件下生成的 DC 相比，其具有增强的免疫原性。此外，我们展示了使用 CCL3 蛋白生成高迁移性 DC 的方法，这些 DC 容易向淋巴器官趋化因子迁移。这两种方法的结合代表了一种简单且临床可行的方法，可生成具有出色迁移能力的高度成熟的 DC。
