Background/Objectives:Dendritic-cell-derived exosomes (DEXs) have demonstrated immunostimulatory potential in cancer immunotherapy, yet their clinical application remains constrained by their cryodependence, compositional heterogeneity, and limited scalability. To address these limitations, we developed an ultrapurified phospholipoproteic complex (PLPC), a dendritic-secretome-derived formulation stabilized through ultracentrifugation and lyophilization that has been engineered to preserve its immunological function and structural integrity.Methods:Secretomes were processed under four conditions (fresh, concentrated, cryopreserved, and lyophilized PLPC) and compared through proteomic and functional profiling. Mass spectrometry (LC-MS/MS) analysis revealed that the PLPC retained a significantly enriched set of immunoregulatory proteins—including QSOX1, CCL22, and SDCBP—and exhibited superior preservation of post-translational modifications.Results:Ex vivo co-culture assays with human peripheral blood mononuclear cells (PBMCs) demonstrated that the PLPC induced robust secretion of IFN-γ, TNF-α, and IL-6 while concurrently suppressing IL-10, achieving an IFN-γ/IL-10 ratio exceeding 3.5. Flow cytometry confirmed the substantial activation of both CD4⁺ and CD8⁺ T cells, while apoptosis assays showed selective tumor cytotoxicity (>55% tumor apoptosis) with minimal impact on non-malignant cells (>92% viability).Conclusions:These findings establish the PLPC as a reproducible, Th1-polarizing immunomodulator with selective antitumor activity, ambient-temperature stability, and compatibility with non-invasive administration. Overall, the PLPC emerges as a scalable, cell-free immunotherapeutic platform with translational potential to reprogram immune suppression in metastatic therapy-resistant cancer settings.
背景/目的：树突状细胞来源的外泌体（DEXs）在癌症免疫治疗中已显示出免疫刺激潜力，但其临床应用仍受限于低温依赖性、成分异质性和有限的可扩展性。为克服这些局限，我们开发了一种超纯化磷脂蛋白复合物（PLPC），这是一种通过超速离心和冻干技术稳定的树突状细胞分泌组衍生制剂，旨在保持其免疫学功能和结构完整性。 方法：分泌组在四种条件下（新鲜、浓缩、冷冻保存和冻干PLPC）进行处理，并通过蛋白质组学和功能谱分析进行比较。质谱分析（LC-MS/MS）显示，PLPC保留了显著富集的免疫调节蛋白（包括QSOX1、CCL22和SDCBP），并表现出优异的翻译后修饰保留能力。 结果：与人外周血单个核细胞（PBMCs）的体外共培养实验表明，PLPC能强力诱导IFN-γ、TNF-α和IL-6的分泌，同时抑制IL-10，使IFN-γ/IL-10比值超过3.5。流式细胞术证实了CD4⁺和CD8⁺ T细胞的显著活化，而细胞凋亡实验显示其对肿瘤细胞具有选择性细胞毒性（肿瘤细胞凋亡率>55%），对非恶性细胞影响极小（存活率>92%）。 结论：这些研究结果确立了PLPC作为一种可重复生产、具有Th1极化特性的免疫调节剂，具备选择性抗肿瘤活性、常温稳定性和非侵入性给药的兼容性。总体而言，PLPC作为一种可扩展的无细胞免疫治疗平台，在转移性耐药癌症治疗中具有重编程免疫抑制的转化潜力。
肿瘤(癌症)患者之家