UOC Reumatologia E Malattie Autoinfiammatorie, IRCCS Istituto Giannina Gaslini, Genoa, Italy.
Dipartimento Di Neuroscienze, Riabilitazione, OftalmologiaGenetica e Scienze Materno-Infantili (DINOGMI), Università Degli Studi Di Genova, Genoa, Italy.
J Clin Immunol. 2024 Jan 17;44(2):49. doi: 10.1007/s10875-023-01649-7.
Syndrome of undifferentiated recurrent fever (SURF) is characterized by recurrent fevers, a lack of confirmed molecular diagnosis, and a complete or partial response to colchicine. Despite the clinical similarities to familial Mediterranean fever (FMF), the underlying inflammatory mechanisms of SURF are not yet understood. We here analyzed the in vitro activation of the pyrin inflammasome in a cohort of SURF patients compared to FMF and PFAPA patients. Peripheral blood mononuclear cells (PBMC) were collected from SURF (both colchicine-treated and untreated), FMF, PFAPA patients, and healthy donors. PBMC were stimulated ex vivo with Clostridium difficile toxin A (TcdA) and a PKC inhibitor (UCN-01), in the presence or absence of colchicine. The assembly of the pyrin inflammasome was evaluated by measuring the presence of apoptosis-associated Speck-like protein containing caspase recruitment domain (ASC) specks in monocytes using flow cytometry. IL-1β secretion was quantified using an ELISA assay. No differences in TcdA-induced activation of pyrin inflammasome were observed among FMF, PFAPA, and healthy donors. Untreated SURF patients showed a reduced response to TcdA, which was normalized after colchicine treatment. In contrast to FMF, SURF patients, similar to PFAPA patients and healthy donors, did not exhibit pyrin inflammasome activation in response to UCN-01-mediated pyrin dephosphorylation. These data demonstrate that in vitro functional analysis of pyrin inflammasome activation can differentiate SURF from FMF and PFAPA patients, suggesting the involvement of the pyrin inflammasome in the pathophysiology of SURF.
未分化复发性发热综合征(SURF)的特征是反复发作的发热、缺乏明确的分子诊断以及对秋水仙碱的完全或部分反应。尽管与家族性地中海热(FMF)具有临床相似性,但 SURF 的潜在炎症机制尚不清楚。我们在此分析了 SURF 患者与 FMF 和 PFAPA 患者相比,体外吡喃炎性小体的激活情况。从 SURF(秋水仙碱治疗和未治疗)、FMF、PFAPA 患者和健康供体中收集外周血单核细胞(PBMC)。在存在或不存在秋水仙碱的情况下,将 PBMC 用艰难梭菌毒素 A(TcdA)和蛋白激酶 C 抑制剂(UCN-01)体外刺激。通过流式细胞术测量单核细胞中凋亡相关斑点样蛋白含有半胱氨酸蛋白酶募集结构域(ASC)斑点的存在来评估吡喃炎性小体的组装。使用 ELISA 测定法定量测定 IL-1β 分泌。在 FMF、PFAPA 和健康供体之间,TcdA 诱导的吡喃炎性小体的激活没有差异。未经治疗的 SURF 患者对 TcdA 的反应减弱,经秋水仙碱治疗后得到正常化。与 FMF 不同,SURF 患者与 PFAPA 患者和健康供体一样,在用 UCN-01 介导的吡喃去磷酸化反应时,不会激活吡喃炎性小体。这些数据表明,体外吡喃炎性小体激活的功能分析可以将 SURF 与 FMF 和 PFAPA 患者区分开来,表明吡喃炎性小体参与了 SURF 的病理生理学。