Analysis of monocyte subsets from pretreatment RCC patients compared to healthy donors.. Analysis of monocyte subsets from pretreatment RCC patients compared to healthy donors.

Peripheral blood monocytes are the starting material utilized in conventional dendritic cell (DC) vaccination for the treatment of a broad range of malignancies. While the use of cytokines and growth factors to polarize monocyte-derived DC to distinct phenotypes is well-established, little is known about the contributions of distinct human monocyte subsets to monocyte-derived DC function and patient responses to vaccination. To investigate the status of monocyte subsets in cancer patients and following culture into DC, we isolated classical (C-Mo), intermediate (I-Mo), and non-classical (NC-Mo) from the peripheral blood of renal cell carcinoma (RCC) patients prior to DC vaccination (NCT00085436) and from anonymous healthy donors. Patients treated with DC vaccination who were long term survivors (>100 months survival) had a unique monocyte signature with a two-fold higher percentage of NC-Mo in pretreatment peripheral blood compared to other RCC patients. RCC patient monocytes from each subset were transcriptionally distinct from healthy donor monocytes. Further transcriptional analysis determined that each monocyte subset was characterized by a discrete gene expression profile before and after DC maturation. Phenotypic analysis showed that DC derived from NC-Mo expressed higher levels of CD80, CD83, CD86, HLA-DR, and CD40 compared to DC originating from C-Mo and secreted increased amounts of IL-12p70 following CD40L stimulation. Collectively, these findings establish that DC derived from NC-Mo are potent antigen presenting cells and provide the foundation for future vaccination strategies that enrich NC-Mo prior to DC maturation. Overall design: Blood monocyte subsets from 8 pretreatment RCC patients were compared to 4 healthy donor controls.

外周血单核细胞是常规树突状细胞（dendritic cell, DC）疫苗疗法治疗多种恶性肿瘤的起始原材料。尽管利用细胞因子与生长因子将单核细胞衍生DC极化为特定功能表型的方法已得到广泛验证，但目前对于人类不同单核细胞亚群对单核细胞衍生DC功能以及患者疫苗应答的具体贡献仍不甚明晰。为探究癌症患者体内以及体外培养为DC后的单核细胞亚群特征，本研究从肾细胞癌（renal cell carcinoma, RCC）患者树突状细胞疫苗接种前（临床试验注册号NCT00085436）以及匿名健康供者的外周血中，分离得到经典型单核细胞（classical monocyte, C-Mo）、中间型单核细胞（intermediate monocyte, I-Mo）与非经典型单核细胞（non-classical monocyte, NC-Mo）。经DC疫苗治疗且生存期超过100个月的长期存活肾细胞癌患者，其预处理外周血中NC-Mo的占比较其他患者高出两倍，呈现独特的单核细胞分子特征。各亚群的肾细胞癌患者单核细胞在转录谱上均与健康供者单核细胞存在显著差异。进一步转录分析表明，在DC成熟前后，各单核细胞亚群均具有独特的基因表达谱。表型分析结果显示，相较于源自C-Mo的DC，由NC-Mo分化获得的DC可高表达CD80、CD83、CD86、HLA-DR及CD40，且在CD40L刺激后可分泌更多IL-12p70。综上，本研究证实由NC-Mo分化而来的DC是强效抗原呈递细胞，为未来在DC成熟前富集NC-Mo的疫苗研发策略奠定了理论基础。整体实验设计：将8名预处理阶段肾细胞癌患者的血液单核细胞亚群与4名健康供者对照进行对比分析。