WS10.01Microbial tolerance in cystic fibrosis airways is imprinted by a new type of immune memory and can be countered by novel host-directed therapy

IF 5.4 2区医学 Q1 RESPIRATORY SYSTEM

Journal of Cystic Fibrosis Pub Date : 2025-06-01 DOI:10.1016/j.jcf.2025.03.546

B. Dobosh , D. Luthra , J. Hosten , P. Kumar , D. Moncada Giraldo , J. Coppinger , R. Tirouvanziam

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引用次数: 0

Abstract

Objectives

Chronic infections in cystic fibrosis (CF) airways are not efficiently countered by antibiotics and CFTR modulators. Our prior studies showed that neutrophils conditioned by the CF airway milieu actively repress microbial killing (Margaroli, Cell Rep Med 2021). Here, we asked if extracellular vesicles (EVs) may underlie this microbial tolerance.

Methods

EVs from CF airway supernatant (CF ASN, obtained from sputum by homogenization and removal of cells, bacteria and debris) were characterized by nanoparticle tracking analysis, Western blot, proteomics, qRT-PCR and RNASeq. CF ASN either whole or EV-depleted was used in a transmigration model to recapitulate airway leukocyte conditioning (Dobosh, STAR Prot 2021). EVs from transmigrated neutrophils were used in sequential transmigrations to evaluate signaling recursivity. Microbial killing was assessed by incubation of leukocytes with Pseudomonas aeruginosa.

Results

Unlike neutrophils recruited to full CF ASN, those recruited to CF ASN devoid of neutrophil EVs showed normal bacterial killing and their EVs had normal content and did not induce bacterial tolerance in the next wave of transmigrated cells. Like neutrophils, monocytes recruited to CF ASN and differentiated into macrophages also repressed bacterial killing. By omics analysis, we identified the long non-coding RNA MALAT1 in neutrophil EVs, and the histone deacetylase HDAC11 in airway leukocytes as key to this pathological loop. Clinically, MALAT1 levels in CF sputum was negatively correlated with lung function. Therapeutically, HDAC11 inhibitor SIS-17 normalized microbial killing by leukocytes in CF ASN and abrogated EV-mediated tolerance imprinting.

Conclusion

Neutrophil EVs mediate a new type of recursive immune memory in CF airways, which forces leukocytes to tolerate microbes and can be countered by host-directed therapy.

Acknowledgments: CF Biospecimen Repository, Emory Cytometry and Genomics Cores; R01HL159058 (NIH), TIROUV22G0-CFRD (CFF).

查看原文本刊更多论文

囊性纤维化气道中的微生物耐受性被一种新的免疫记忆所印记，并且可以通过新的宿主定向治疗来对抗

目的囊性纤维化气道慢性感染不能通过抗生素和CFTR调节剂有效对抗。我们之前的研究表明，CF气道环境调节的中性粒细胞积极抑制微生物杀灭（Margaroli, Cell Rep Med 2021）。在这里，我们询问细胞外囊泡（EVs）是否可能是这种微生物耐受性的基础。方法采用纳米颗粒跟踪分析、Western blot、蛋白质组学、qRT-PCR和RNASeq等方法对CF气道上清（CF ASN）的sev进行表征。在转运模型中使用完整或ev耗尽的CF ASN来重现气道白细胞调节（Dobosh, STAR Prot 2021）。来自迁移中性粒细胞的ev被用于序列迁移来评估信号递归性。用铜绿假单胞菌培养白细胞来评估微生物杀灭效果。结果与招募到完整CF ASN的中性粒细胞一样，招募到缺乏中性粒细胞ev的CF ASN的中性粒细胞表现出正常的细菌杀伤，其ev含量正常，并且在下一波转移细胞中不诱导细菌耐受。与中性粒细胞一样，单核细胞被募集到CF ASN并分化为巨噬细胞，也抑制了细菌的杀伤。通过组学分析，我们发现中性粒细胞ev中的长链非编码RNA MALAT1和气道白细胞中的组蛋白去乙酰化酶HDAC11是这一病理循环的关键。临床上，CF痰中MALAT1水平与肺功能呈负相关。在治疗上，HDAC11抑制剂SIS-17使CF ASN中白细胞的微生物杀伤正常化，并消除了ev介导的耐受印迹。结论在CF气道中，中性粒细胞介导一种新的递归免疫记忆，迫使白细胞耐受微生物，可通过宿主定向治疗进行对抗。致谢：CF生物标本库，Emory细胞术和基因组学核心；R01hl159058 (nih), tirouv22g0-cfrd （cff）。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Cystic Fibrosis 医学-呼吸系统

CiteScore

10.10

自引率

13.50%

发文量

1361

审稿时长

50 days

期刊介绍： The Journal of Cystic Fibrosis is the official journal of the European Cystic Fibrosis Society. The journal is devoted to promoting the research and treatment of cystic fibrosis. To this end the journal publishes original scientific articles, editorials, case reports, short communications and other information relevant to cystic fibrosis. The journal also publishes news and articles concerning the activities and policies of the ECFS as well as those of other societies related the ECFS.