Introduction: Monitoring the therapeutic response of pancreatic ductal adenocarcinoma (PDAC) patients is crucial to determine treatment strategies. Several studies have examined the effectiveness of FOLFIRINOX as a first-line treatment in patients with locally advanced pancreatic cancer, but little attention has been paid to the immunologic alterations in peripheral blood caused by this chemotherapy regimen. Furthermore, the influence of the measurement type (e.g., flow cytometry and targeted gene expression) on the clinical discoveries is unknown. Therefore, we aimed to scrutinize the influence of using flow cytometry or targeted immune gene expression to study the immunological changes in blood samples of PDAC patients who were treated with a single-cycle FOLFIRINOX combined with lipegfilgrastim (FFX-Lipeg). Material and Methods: Whole-blood samples from 44 PDAC patients were collected at two time points: before the first FOLFIRINOX cycle and 14 days after the first cycle. EDTA blood tubes were used for multiplex flow cytometry analyses to quantify 18 immune cell populations and for complete blood count tests as the standard clinical routine. The flow cytometry data were analyzed with FlowJo software. In addition, Tempus blood tubes were used to isolate RNA and measure 1230 immune-related genes using NanoString Technology®. Data quality control, normalization, and analysis were performed using nSolver™ software and the Advanced Analysis module. Results: FFX-Lipeg treatment increased the number of neutrophils and monocytes, as shown by flow cytometry and complete blood count in concordance with elevated gene expression measured via targeted gene expression profiling analysis. Interestingly, flow cytometry analysis showed an increase in the number of B and T cells after treatment, while targeted gene expression analysis showed a decrease in B and T cell-specific gene expression. Conclusions: Targeted gene expression complements flow cytometry analysis to provide a comprehensive understanding of the effects of FFX-Lipeg. Flow cytometry and targeted gene expression showed increases in neutrophils and monocytes after FFX-Lipeg. The number of lymphocytes is increased after treatment; nevertheless, their cell-specific gene expression levels are downregulated. This highlights that different techniques influence clinical discoveries. Therefore, it is important to carefully select the measurement technique used to study the effect of a treatment.
引言：监测胰腺导管腺癌（PDAC）患者的治疗反应对于确定治疗策略至关重要。多项研究已探讨FOLFIRINOX方案作为局部晚期胰腺癌患者一线治疗的有效性，但该化疗方案引起的外周血免疫学改变却鲜受关注。此外，检测方法（如流式细胞术与靶向基因表达分析）对临床发现的影响尚不明确。因此，本研究旨在系统分析使用流式细胞术或靶向免疫基因表达技术，研究接受单周期FOLFIRINOX联合利培格司亭（FFX-Lipeg）治疗的PDAC患者血液样本免疫学变化时产生的影响差异。 材料与方法：采集44例PDAC患者两个时间点的全血样本：首次FOLFIRINOX周期前及周期后第14天。采用EDTA抗凝管进行多重流式细胞术分析以量化18个免疫细胞群，并行全血细胞计数作为标准临床常规检测。流式细胞术数据使用FlowJo软件分析。同时采用Tempus采血管分离RNA，通过NanoString Technology®检测1230个免疫相关基因表达。数据质量控制、标准化及分析使用nSolver™软件及其高级分析模块完成。 结果：FFX-Lipeg治疗后，流式细胞术与全血细胞计数均显示中性粒细胞和单核细胞数量增加，这与靶向基因表达谱分析检测到的基因表达上调结果一致。值得注意的是，流式细胞术分析显示治疗后B细胞和T细胞数量增加，而靶向基因表达分析却显示B细胞和T细胞特异性基因表达下调。 结论：靶向基因表达分析可对流式细胞术形成有效补充，为全面理解FFX-Lipeg的治疗效应提供多维视角。两种技术均证实FFX-Lipeg治疗后中性粒细胞和单核细胞增多。虽然治疗后淋巴细胞数量增加，但其细胞特异性基因表达水平却呈现下调。这凸显了不同检测技术对临床发现的重要影响，因此在研究治疗方案效果时需审慎选择检测方法。
肿瘤(癌症)患者之家