Although clinically effective, the actions of IFNα, either produced endogenously or by therapeutic delivery, remain poorly understood. Emblematic of this research gap is the disparate array of notable side effects that occur in susceptible individuals, such as neuropsychiatric consequences, autoimmune phenomena, and infectious complications. We hypothesised that these complications are driven at least in part by dysregulated cellular metabolism. Male Wistar rats were treated with either 170,000 IU/kg human recombinant IFNα-2a or BSA/saline (0.9% NaCl) three times per week for three weeks. Bone marrow (BM) immune cells were isolated from the excised femurs for glycolytic rate and mitochondrial function assessment using Agilent Seahorse Technology. Frequencies of immune cell populations were assessed by flow cytometry to determine whether leukopoietic changes had occurred in both blood and BM. Plasma levels of lactate and succinate were also determined. BMDMs were metabolically assessed as above, as well as their metabolic response to an antigenic stimulus (iH37Rv). We observed that BM immune cells from IFN-treated rats exhibit a hypermetabolic state (increased basal OCR/GlycoPER) with decreased mitochondrial metabolic respiration and increased non-mitochondrial OCR. Flow cytometry results indicated an increase in immature granulocytes (RP1- SSChi CD45lo) only in the blood, together with increased succinate levels in the plasma. BMDMs from IFN-treated rats retained the hypermetabolic phenotype after differentiation and failed to induce a step-up in glycolysis and mitochondrial respiration after bacterial stimulation. This work provides the first evidence of the effects of IFNα treatment in inducing hypermetabolic immune features that are associated with markers of inflammation, leukopoiesis, and defective responses to bacterial stimulation.

尽管临床上有效，但对于内源性或通过治疗递送产生的 IFNα 的作用仍知之甚少。这一研究空白的标志是易感个体中发生的一系列不同的显着副作用，例如神经精神后果、自身免疫现象和感染并发症。我们假设这些并发症至少部分是由细胞代谢失调引起的。雄性 Wistar 大鼠每周 3 次，用 170,000 IU/kg 人重组 IFNα-2a 或 BSA/盐水 (0.9% NaCl) 治疗，持续三周。使用安捷伦海马技术从切除的股骨中分离骨髓 (BM) 免疫细胞，用于糖酵解速率和线粒体功能评估。通过流式细胞术评估免疫细胞群的频率，以确定血液和骨髓中是否发生白细胞生成变化。还测定了乳酸和琥珀酸的血浆水平。如上所述对 BMDM 及其对抗原刺激 (iH37Rv) 的代谢反应进行代谢评估。我们观察到，来自 IFN 治疗的大鼠的 BM 免疫细胞表现出代谢亢进状态（基础 OCR/GlycoPER 增加），线粒体代谢呼吸减少，非线粒体 OCR 增加。流式细胞术结果表明，仅血液中未成熟粒细胞（RP1-SSChi CD45lo）增加，并且血浆中琥珀酸盐水平增加。来自经 IFN 处理的大鼠的 BMDM 在分化后保留了代谢亢进的表型，并且在细菌刺激后未能诱导糖酵解和线粒体呼吸的增强。这项工作提供了第一个证据，证明 IFNα 治疗可诱导代谢亢进的免疫特征，这些特征与炎症、白细胞生成和对细菌刺激的反应缺陷等标志物相关。