The Role of IL-17 in fungal keratitis

IL-17在真菌性角膜炎中的作用

• 批准号: 8521211
• 负责人: Patricia R Taylor
• 金额: $ 5.39万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8521211
• 关键词: Adoptive Transfer; Antifungal Agents; Aspergillus; Biological Assay; Blindness; Bone Marrow; C57BL/6 Mouse; CD4 Positive T Lymphocytes; Cells; Cornea; Corneal Diseases; Corneal Opacity; Corneal Stroma; Corneal Ulcer; Data; Development; Disease; EMSA; Flow Cytometry; Fusarium; Hyphae; IL8RB gene; Immune response; Immunity; Immunization; In Vitro; Incubated; Infection; Interferons; Interleukin-16; Interleukin-17; Interleukin-6; Keratitis; Lead; Manuscripts; Mediating; Messenger RNA; Modeling; Mus; Mycoses; Nuclear Extract; Phenotype; Population; Production; Promoter Regions; RNA; Reactive Oxygen Species; Recombinants; Recruitment Activity; Regulation; Relative (related person); Role; Route; STAT3 gene; Sorting - Cell Movement; Source; Spleen; Splenocyte; Th1 Cells; Time; Transcript; Visual impairment; Western Blotting; abstracting; adaptive immunity; cell type; chemokine; interleukin-23; killings; mRNA Expression; magnetic beads; neutralizing antibody; neutrophil; novel; research study; subcutaneous

DESCRIPTION (provided by applicant): IL-17 and Th17 cells have an important role in the host response to fungal infections. In the current study, we examined the role of IL-17 in a murine model of Aspergillus and Fusarium infections of the cornea, which are a major cause of blindness and visual impairment worldwide. C57BL/6 mice immunized by intratracheal or subcutaneous routes. Mice immunized by either route demonstrated rapid fungal clearance and significantly reduced corneal disease compared with unimmunized, infected mice which severe corneal opacity and a persistent fungal presence. Intracellular flow cytometry showed development of fungal-specific Th1 and Th17 cells in the spleen following immunization, which were selectively recruited to the cornea after infection (Th17 cells at 48h, and Th1 cells at 72h). Neutralization of IL-17 following immunization, or infection of IL-17-/- mice inhibited the protective phenotype, whereas IFN-? neutralization had no effect. A discrete population of IL-17 producing neutrophils was detected in the corneas 24h after infection corneas of immunized, but not unimmunized mice, and were detected in the spleen and bone marrow. Further, neutrophil depletion impaired IL-17 production at this time and blocked protective immunity. IL-17 producing neutrophils produced elevated ROS and had increased capacity to kill hyphae in vitro, indicating that this population of cells has an important role in regulating fungal infectio. Quantitative PCR of the FACS sorted IL-17 neutrophils from the cornea, spleen and bone marrow showed IL-17 transcripts, indicating de novo synthesis. Further, naive bone marrow neutrophils expressed IL-17 RNA when incubated with splenocyte supernatants from immunized mice, and expression was ablated in the presence of both anti-IL-6 and anti-IL-23. Conversely, stimulation with both IL-16 and IL-23, but not with single Abs stimulated IL-17 expression. Together, these data indicate that IL-17 producing neutrophils are generated by IL-6 and IL-23 in immunized mice, and are recruited to fungal infected corneas, resulting in rapid fungal clearance possibly due to elevated production of reactive oxygen species or interactions with Th17 cells. These findings show a role for IL-17, Th17, and IL-17 producing neutrophils in regulating fungal infections. In the proposed experiments I will determine the relative contribution of Th17 cells and IL-17 producing cells in protective immunity in fungal keratitis. )

描述（由申请人提供）：IL-17和TH17细胞在宿主对真菌感染的反应中具有重要作用。在当前的研究中，我们研究了IL-17在角膜曲霉和镰刀菌感染的鼠模型中的作用，这是世界范围内失明和视觉障碍的主要原因。通过气管内或皮下途径免疫的C57BL/6小鼠。与未免疫的，受感染的小鼠相比，通过任何一条路线免疫的小鼠都表现出快速的真菌清除，并且显着降低了角膜疾病，而这些小鼠严重的角膜不透明和持续的真菌存在。免疫后，细胞内流式细胞术显示脾脏在脾脏中的真菌特异性Th1和Th17细胞的发展，这些细胞在感染后选择性地募集到角膜（48H时Th17细胞，而在72H时Th1细胞）。 免疫后IL-17的中和或IL-17 - / - 小鼠感染抑制了保护性表型，而IFN-？中和没有作用。 在感染免疫的角膜后，在24小时内检测到IL-17产生嗜中性粒细胞的离散群体，但未在脾脏和骨髓中检测到无免疫小鼠。此外，嗜中性粒细胞耗竭损害了IL-17的产生，并阻止了保护性免疫。 IL-17产生嗜中性粒细胞的ROS升高，并具有增加体外杀死菌丝的能力，这表明该细胞群在调节真菌感染中具有重要作用。来自角膜，脾和骨髓的FACS的定量PCR显示了IL-17转录本，表明从头合成。此外，当与免疫小鼠的脾细胞上清液一起孵育时，幼稚的骨髓嗜中性粒细胞表达IL-17 RNA，并且在抗IL-6和抗IL-23的情况下表达消融。相反，IL-16和IL-23均具有刺激，但单个ABS刺激IL-17表达却没有。总之，这些数据表明，IL-17产生嗜中性粒细胞是由IL-6和IL-23在免疫小鼠中产生的，并被募集到被真菌感染的角膜中，从而导致了快速的真菌清除率，可能是由于活性氧的产生升高而导致活性氧或与TH17细胞相互作用。这些发现在调节真菌感染中产生嗜中性粒细胞的IL-17，TH17和IL-17的作用。在拟议的实验中，我将确定Th17细胞和IL-17产生细胞在真菌角膜炎中保护性免疫中的相对贡献。 ）