Objective and design

This study tested the hypothesis that sickle red blood cell (SS-RBC) can induce inflammasome NLRP3 components gene expression in peripheral blood mononuclear cells (PBMCs) as well as interleukin-1β (IL-1β) and leukotriene B4 (LTB4) production. Additionally, we investigated the effect of hydroxyurea (HU) treatment in these inflammatory markers.

Methods

PBMCs from healthy donors (AA-PBMC) were challenged with intact and lysed RBCs from SCA patients (SS-RBC) and from healthy volunteers (AA-RBC). NLRP3, IL-1β, IL-18 and Caspase-1 gene expression levels were assessed by quantitative PCR (qPCR). IL-1β protein levels and LTB4 were measured by ELISA.

Results

We observed that lysed SS-RBC induced the expression of inflammasome NLRP3 components, but this increase was more prominent for CASP1 and IL18 expression levels. Moreover, we observed that intact SS-RBC induced higher production of IL-1β and LTB4 than lysed SS-RBC. Although SCA patients treated with HU have a reduction in NLRP3 gene expression and LTB4 production, this treatment did not modulate the expression of other inflammasome components or IL-1β production.

Conclusions

Thus, our data suggest that caspase-1, IL-1β and IL-18 may contribute to the inflammatory status observed in SCA and that HU treatment may not interfere in this inflammatory pathway.

中文翻译：

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裂解和非裂解镰状红细胞对单核细胞NLRP3炎性体激活和LTB4产生的影响

目标和设计

本研究检验了镰状红细胞 (SS-RBC) 可以诱导外周血单核细胞 (PBMC) 中炎性体 NLRP3 成分基因表达以及白细胞介素 1β (IL-1β) 和白三烯 B4 (LTB4) 产生的假设。此外，我们研究了羟基脲 (HU) 治疗对这些炎症标志物的影响。

方法

来自健康供体 (AA-PBMC) 的 PBMC 受到来自 SCA 患者 (SS-RBC) 和健康志愿者 (AA-RBC) 的完整和裂解的 RBC 的挑战。通过定量 PCR (qPCR) 评估NLRP3、IL-1β、IL-18和Caspase-1基因表达水平。通过ELISA测量IL-1β蛋白水平和LTB4。

结果

我们观察到裂解的 SS-RBC 诱导了炎性体 NLRP3 成分的表达，但这种增加对于CASP1和IL18表达水平更为突出。此外，我们观察到完整的 SS-RBC 比裂解的 SS-RBC 诱导更高的 IL-1β 和 LTB4 产生。尽管 HU 治疗的 SCA 患者NLRP3基因表达和 LTB4 产生减少，但这种治疗并未调节其他炎性体成分的表达或 IL-1β 的产生。

结论

因此，我们的数据表明 caspase-1、IL-1β 和 IL-18 可能导致 SCA 中观察到的炎症状态，并且 HU 治疗可能不会干扰这种炎症途径。