Department of Immunology, University of Sao Paulo, Brazil.
Department of Rheumatology and Clinical Immunology, University of Lübeck, Germany.
J Infect Dis. 2017 Dec 19;216(12):1623-1634. doi: 10.1093/infdis/jix522.
Mutations in genes affecting interferon-γ (IFN-γ) immunity have contributed to understand the role of IFN-γ in protection against intracellular pathogens. However, inborn errors in STAT4, which controls interleukin-12 (IL-12) responses, have not yet been reported. Our objective was to determine the genetic defect in a family with a history of paracoccidioidomycosis.
Genetic analysis was performed by whole-exome sequencing and Sanger sequencing. STAT4 phosphorylation (pSTAT4) and translocation to the nucleus, IFN-γ release by patient lymphocytes, and microbicidal activity of patient monocytes/macrophages were assessed. The effect on STAT4 function was evaluated by site-directed mutagenesis using a lymphoblastoid B cell line (B-LCL) and U3A cells.
A heterozygous missense mutation, c.1952 A>T (p.E651V) in STAT4 was identified in the index patient and her father. Patient's and father's lymphocytes showed reduced pSTAT4, nuclear translocation, and impaired IFN-γ production. Mutant B-LCL and U3A cells also displayed reduced pSTAT4. Patient's and father's peripheral blood mononuclear cells and macrophages demonstrated impaired fungicidal activity compared with those from healthy controls that improved in the presence of recombinant human IFN-γ, but not rhIL-12.
Our data suggest autosomal dominant STAT4 deficiency as a novel inborn error of IL-12-dependent IFN-γ immunity associated with susceptibility to paracoccidioidomycosis.
影响干扰素-γ(IFN-γ)免疫的基因突变有助于了解 IFN-γ在抵抗细胞内病原体中的作用。然而,控制白细胞介素-12(IL-12)反应的 STAT4 中的先天缺陷尚未报道。我们的目的是确定一个有副球孢子菌病病史的家族的遗传缺陷。
通过全外显子组测序和 Sanger 测序进行基因分析。评估 STAT4 磷酸化(pSTAT4)和核易位、患者淋巴细胞释放 IFN-γ以及患者单核细胞/巨噬细胞的杀菌活性。通过使用淋巴母细胞 B 细胞系(B-LCL)和 U3A 细胞进行定点诱变来评估对 STAT4 功能的影响。
在索引患者及其父亲中发现了 STAT4 中的杂合错义突变 c.1952 A>T(p.E651V)。患者和父亲的淋巴细胞显示出减少的 pSTAT4、核易位和 IFN-γ产生受损。突变的 B-LCL 和 U3A 细胞也显示出减少的 pSTAT4。与健康对照相比,患者和父亲的外周血单核细胞和巨噬细胞显示出杀菌活性受损,在重组人 IFN-γ存在的情况下改善,但在 rhIL-12 存在的情况下没有改善。
我们的数据表明,常染色体显性 STAT4 缺陷是一种新的 IL-12 依赖性 IFN-γ免疫先天缺陷,与副球孢子菌病易感性相关。