PO.TB04.02 · 肿瘤生物学
利用41重人源化小鼠免疫检测板对第二代人源化NOG小鼠进行稳健的多重细胞因子分析：跨供体验证及对转化肿瘤学研究的意义
Robust multiplex cytokine profiling in second-generation humanized NOG mice using a 41-plex humanized mouse immune panel: Cross-donor validation and implications for translational oncology studies
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：第二代人源化NOG小鼠（HIS小鼠）可在生理相关的人类免疫微环境中评估免疫肿瘤学机制。然而，临床前疗效研究的转化解读需要对人类细胞因子进行高度特异且可重复的定量。为支持稳健的免疫监测，我们共同评估了一种41重人源化小鼠细胞因子/趋化因子Luminex检测板在不同人类CD34+供体来源及多种HIS NOG模型变体中的性能。
研究目的/假设：我们假设MILLIPLEX®41重人源化小鼠多重检测中的人类特异性分析物将表现出（i）对人类细胞因子的高特异性，且与宿主小鼠分析物的交叉反应最小；（ii）在供体间变异中具有可重复的定量能力；以及（iii）在支持转化肿瘤学应用的先进HIS NOG模型中具有一致的检测灵敏度。
结果：从由≥3个无关CD34+供体池重建、代表多种第二代人源化平台的HIS NOG队列中采集了血浆样本。在所有供体和模型中，人类限制性分析物均产生了高于背景的可定量信号。仅含小鼠的对照样本证实交叉反应可忽略不计。检测间和检测内变异系数（CV）证明了技术可重复性。供体特异性细胞因子模式（如Th1/Th2平衡、髓系相关趋化因子）在各模型间得以保留，表明其反映的是生物学保真度而非检测漂移。基线细胞因子结构与重建动力学及人类免疫细胞亚群频率相关，支持生物学一致性。
结论：这些验证数据表明，MILLIPLEX®41重Luminex检测在第二代HIS NOG小鼠中提供了可重复、供体一致且人类特异的细胞因子定量。这些数据支持该检测适用于肿瘤学研究中需要高分辨率人类细胞因子/趋化因子分析的机制性和药效学读数。可靠的多重细胞因子测量强化了HIS小鼠研究与人类免疫肿瘤学反应之间的转化桥梁，有助于改善对治疗机制、生物标志物及治疗诱导的免疫调节的解读。
查看英文原文 English abstract
Background: Second-generation humanized NOG mice (HIS mice) enable evaluation of immune-oncology mechanisms within a physiologically relevant human immune microenvironment. However, translational interpretation of preclinical efficacy studies requires highly specific and reproducible quantification of human cytokines. To support robust immune monitoring, we co-evaluated a 41-plex humanized mouse cytokine/chemokine Luminex panel for performance across diverse human CD34+ donor sources and multiple HIS NOG model variants.
Study Aims/Hypothesis: We hypothesized that the human-specific analytes in the MILLIPLEX® 41-plex humanized mouse multiplex assay would show (i) high specificity for human cytokines with minimal cross-reactivity to host murine analytes, (ii) reproducible quantification across donor-to-donor variation, and (iii) consistent detection sensitivity across advanced HIS NOG models supporting translational oncology applications.
Results: Plasma samples were collected from HIS NOG cohorts reconstituted with ≥3 unrelated CD34+ donor pools and representing multiple second-generation humanized platforms. Across all donors and models, human-restricted analytes yielded quantifiable signals above background. Murine-only control samples confirmed negligible cross-reactivity. Inter-assay and intra-assay CVs demonstrated technical reproducibility. Donor-specific cytokine patterns (e.g., Th1/Th2 balance, myeloid-associated chemokines) were preserved across models, indicating biological fidelity rather than assay drift. Baseline cytokine architecture correlated with reconstitution kinetics and human immune cell subset frequencies, supporting biological coherence.
Conclusions: These validation data demonstrate that the MILLIPLEX® 41-plex Luminex assay provides reproducible, donor-consistent, and human-specific cytokine quantification in second-generation HIS NOG mice. These data support the assay's suitability for mechanistic and pharmacodynamic readouts in oncology studies requiring high-resolution human cytokine/chemokine profiling. Reliable multiplex cytokine measurement strengthens the translational bridge between HIS mouse studies and human immuno-oncology responses, enabling improved interpretation of therapeutic mechanisms, biomarkers, and treatment-induced immune modulation.
利益披露 Disclosure
P. Dubé, None..
B. Gilliam, None..
A. Bell, None..
N. Smith, None..
J. Richardson, None..
T. Raeber, None..
S. Curran, None..
M. Buczek, None.