Human neutrophils activated by TLR8 agonists, with or without IFNγ, synthesize and release EBI3, but not IL-12, IL-27, IL-35, or IL-39
Human neutrophils activated by TLR8 agonists, with or without IFNγ, synthesize and release EBI3, but not IL-12, IL-27, IL-35, or IL-39
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DOI：
10.1002/jlb.3ma0520-054r
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发表时间：
2020-06-01
影响因子：
5.5
通讯作者：
Tamassia, Nicola
中科院分区：
文献类型：
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作者：
Cassatella, Marco A.;Gardiman, Elisa;Tamassia, Nicola
IL-12家族细胞因子在先天免疫和获得性免疫中发挥重要作用。这些细胞因子包括具有两个β(IL-12B和EB病毒诱导基因3[EBI3])链的异二聚体，分别共享IL-12B(IL-12B+IL-12A)和IL-23(IL-12B+IL-23A)、IL-27(EBI3+IL-27A)、IL-35(EBI3+IL-12A)和IL-39(EBI3+IL-23A)。在此背景下，我们最近报道，与TLR8激动剂孵育的高纯度中性粒细胞产生功能性IL-23。此前，我们发现中性粒细胞与内毒素+干扰素-γ孵育20小时后产生IL-12。在这里，我们研究了高纯度、TLR8激活的中性粒细胞是否产生EBI3，进而产生含有EBI3的IL-12成员IL-27、IL-35和IL-39。我们报道，中性粒细胞与TLR8配体、肿瘤坏死因子α以及较少程度的脂多糖孵育后，产生和释放显著数量的EBI3，但不产生IL-27A，因此排除了产生IL-27的可能性。我们还报告了一系列不成功的实验，以调查中性粒细胞来源的EBI3是否与IL-23A结合形成IL-39。此外，我们还发现，与之前的发现相反，与TLR8或TLR4配体共同孵育的中性粒细胞既不表达/产生IL-12，也不产生IL-35，这是因为与先前的发现相反，干扰素不能激活IL-12A转录。甚至在干扰素-伽马加R848处理的中性粒细胞的培养上清液中也检测不到IL-27，尽管它们被发现积累了IL27A转录本。最后，通过免疫组织化学实验，EBI3阳性的中性粒细胞仅在不同的病理组织中被发现，包括憩室炎、胆囊炎、Gorham病和亨氏巴尔通体感染，这意味着中性粒细胞来源的EBI3在体内具有特定的作用。
The IL-12 family of cytokines plays crucial functions in innate and adaptive immunity. These cytokines include heterodimers sharing distinct alpha (IL-12A, IL-23A, and IL-27A) with two beta (IL-12B and Epstein-Barr virus induced gene 3 [EBI3]) chains, respectively, IL-12 (IL-12B plus IL-12A) and IL-23 (IL-12B plus IL-23A) sharing IL-12B, IL-27 (EBI3 plus IL-27A), IL-35 (EBI3 plus IL-12A), and IL-39 (EBI3 plus IL-23A) sharing EBI3. In this context, we have recently reported that highly pure neutrophils incubated with TLR8 agonists produce functional IL-23. Previously, we showed that neutrophils incubated with LPS plus IFN gamma for 20 h produce IL-12. Herein, we investigated whether highly pure, TLR8-activated, neutrophils produce EBI3, and in turn IL-27, IL-35, and IL-39, the IL-12 members containing it. We report that neutrophils incubated with TLR8 ligands, TNF alpha and, to a lesser extent, LPS, produce and release remarkable amounts of EBI3, but not IL-27A, consequently excluding the possibility for an IL-27 production. We also report a series of unsuccessful experiments performed to investigate whether neutrophil-derived EBI3 associates with IL-23A to form IL-39. Furthermore, we show that neutrophils incubated with IFN gamma in combination with either TLR8 or TLR4 ligands express/produce neither IL-12, nor IL-35, due to the inability of IFN gamma, contrary to previous findings, to activate IL12A transcription. Even IL-27 was undetectable in supernatants harvested from IFN gamma plus R848-treated neutrophils, although they were found to accumulate IL27A transcripts. Finally, by immunohistochemistry experiments, EBI3-positive neutrophils were found in discrete pathologies only, including diverticulitis, cholecystitis, Gorham disease, and Bartonella Henselae infection, implying a specific role of neutrophil-derived EBI3 in vivo.