We aimed to use a novel standardized whole-blood stimulation system to evaluate differences in the functional immune reconstitution in patients early after allogeneic haematopoietic cell transplantation (HCT). Between April and September 2018, 30 patients undergoing HCT had whole blood samples collected around day −21 (day 0 being the day of haematopoietic cell infusion) and day +28. Whole blood was transferred to TruCulture assays comprising prefilled incubation tubes with cell culture medium and a standardized stimulus. We used a panel of four stimuli (lipopolysaccharide, resiquimod, heat-killed Candida albicans and polyinosinic:polycytidylic acid) and a blank, designed to evaluate the function of critical extra- and intracellular immunological signalling pathways. For each stimulus, the cytokine response was assessed by the concentration of interferon-γ, interleukin (IL)-12p40, IL-10, IL-1β, IL-6, IL-8, IL-10, IL-12p40, IL-17A and tumour necrosis factor-α using a multiplex Luminex assay. Pre-HCT cytokine responses were globally decreased across several different stimuli. Despite patients receiving immunosuppressive prophylaxis at the time, post-HCT cytokine responses were higher and less intercorrelated than pre-HCT responses, also after adjusting for differences in the leukocyte differential counts. For the resiquimod and heat-killed Candida albicans stimuli, we identified a cluster of patients in whom post-HCT responses were lower than average across several cytokines, indicating a possible functional immune deficiency. Our findings suggest that the standardized whole blood stimulation system can be used to reveal heterogeneity in the in vitro cytokine responses to various stimuli after HCT. Larger studies are needed to address if the functional immune reconstitution after HCT can predict the risk of infections.

中文翻译：

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异基因造血细胞移植后早期功能性免疫重建：受刺激全血中移植前和移植后细胞因子反应的比较

我们旨在使用一种新的标准化全血刺激系统来评估异基因造血细胞移植 (HCT) 后早期患者功能性免疫重建的差异。2018 年 4 月至 9 月期间，30 名接受 HCT 的患者在第 -21 天（第 0 天是造血细胞输注日）和第 +28 天前后收集了全血样本。全血被转移到 TruCulture 检测中，该检测包含预装了细胞培养基和标准化刺激物的孵育管。我们使用了一组四种刺激物（脂多​​糖、瑞喹莫特、热灭活的白色念珠菌和聚肌苷酸：聚胞苷酸）和一个空白，旨在评估关键的细胞外和细胞内免疫信号通路的功能。对于每个刺激，通过干扰素-γ、白细胞介素 (IL)-12p40、IL-10、IL-1β、IL-6、IL-8、IL-10、IL-12p40、IL-17A 和肿瘤坏死的浓度评估细胞因子反应因子-α 使用多重 Luminex 检测。Pre-HCT 细胞因子反应在几种不同的刺激下整体下降。尽管患者当时接受了免疫抑制预防，但在调整了白细胞分类计数差异后，HCT 后细胞因子反应比 HCT 前反应更高且相关性更小。对于瑞喹莫特和热灭活的白色念珠菌刺激物，我们确定了一组患者的 HCT 后反应低于多种细胞因子的平均值，表明可能存在功能性免疫缺陷。我们的研究结果表明，标准化的全血刺激系统可用于揭示 HCT 后体外细胞因子对各种刺激的反应的异质性。需要更大规模的研究来解决 HCT 后功能性免疫重建是否可以预测感染风险。