Trans-epithelial migration is essential for neutrophil activation during RSV infection

作者全名："Robinson, Elisabeth; Herbert, Jenny Amanda; Palor, Machaela; Ren, Luo; Larken, Isobel; Patel, Alisha; Moulding, Dale; Cortina-Borja, Mario; Smyth, Rosalind Louise; Smith, Claire Mary"

作者地址："[Robinson, Elisabeth; Herbert, Jenny Amanda; Palor, Machaela; Ren, Luo; Larken, Isobel; Patel, Alisha; Moulding, Dale; Cortina-Borja, Mario; Smyth, Rosalind Louise; Smith, Claire Mary] UCL Great Ormond St Inst Child Hlth, Infect Immun & Inflammat Dept, 30 Guilford St, London WC1N1EH, England; [Herbert, Jenny Amanda] Univ Manchester, Fac Biol Med & Hlth, Sch Med Sci, Oxford Rd, Manchester M13 9PL, Lancs, England; [Ren, Luo] Chongqing Med Univ, Childrens Hosp, Dept Resp Med, Chongqing 400014, Peoples R China; [Smith, Claire Mary] UCL Great Ormond St Inst Child Hlth, 30 Guilford St, London WC1N 1EH, England"

通信作者："Smith, CM (通讯作者)，UCL Great Ormond St Inst Child Hlth, 30 Guilford St, London WC1N 1EH, England."

来源：JOURNAL OF LEUKOCYTE BIOLOGY

ESI学科分类：IMMUNOLOGY

WOS号：WOS:000960220600003

JCR分区：Q2

影响因子：3.6

年份：2023

卷号：113

期号：4

开始页：354

结束页：364

文献类型：Article

关键词：virus infection; bronchiolitis; airway epithelial cells; migration; movement; neutrophils

摘要："Building on previous work of neutrophil function, we propose 3 main phases of early neutrophil recruitment and behavior in the airways during RSV infection. Phase 1: Initial chemotaxis and adherence: here unstimulated circulating neutrophils expressing baseline levels of CD11b migrate across infected AECs in response to chemotactic signals in the apical supernatant. Some neutrophils remain adherent to the infected AECs. Phase 2: Activation and reverse migration: once on the apical side of the epithelium, neutrophils increase expression of CD11b and other activation-associated markers, and some ""activated"" neutrophils undergo reverse migration. Neutrophils with greater expression of CD11b are detected on the basolateral side. Phase 3: Amplified chemotaxis and clustering: after 20 min, adherent neutrophils begin to rapidly cluster on RSV-infected primary airway epithelial cells cultures, mediated by signaling from a dying neutrophil. Drawing created using BioRender.com. The recruitment of neutrophils to the infected airway occurs early following respiratory syncytial virus (RSV) infection, and high numbers of activated neutrophils in the airway and blood are associated with the development of severe disease. The aim of this study was to investigate whether trans-epithelial migration is sufficient and necessary for neutrophil activation during RSV infection. Here, we used flow cytometry and novel live-cell fluorescent microscopy to track neutrophil movement during trans-epithelial migration and measure the expression of key activation markers in a human model of RSV infection. We found that when migration occurred, neutrophil expression of CD11b, CD62L, CD64, NE, and MPO increased. However, the same increase did not occur on basolateral neutrophils when neutrophils were prevented from migrating, suggesting that activated neutrophils reverse migrate from the airway to the bloodstream side, as has been suggested by clinical observations. We then combined our findings with the temporal and spatial profiling and suggest 3 initial phases of neutrophil recruitment and behavior in the airways during RSV infection; (1) initial chemotaxis; (2) neutrophil activation and reverse migration; and (3) amplified chemotaxis and clustering, all of which occur within 20 min. This work and the novel outputs could be used to develop therapeutics and provide new insight into how neutrophil activation and a dysregulated neutrophil response to RSV mediates disease severity."

基金机构：Academy of Medical Science [NIF004/1012]; Great Ormond Street Children's Charity [W1802]; Animal Free Research UK [AFR19-20274]; BBSRC [BB/V006738/1]; GOSH Children's Charity [COVID_CSmith_017]; Wellcome Trust [212516/Z/18/Z]; NIHR Great Ormond Street Hospital Biomedical Research Centre; NIHR GOSH BRC award [17DD08]

基金资助正文："LR was a recipient of a Newton fellowship from the Academy of Medical Science (ref. NIF004/1012). RLS was supported by the Great Ormond Street Children's Charity (grant code W1802). CMS is a recipient of grants from Animal Free Research UK (AFR19-20274), BBSRC (BB/V006738/1), GOSH Children's Charity (COVID_CSmith_017), and the Wellcome Trust (212516/Z/18/Z). This research was supported by the NIHR Great Ormond Street Hospital Biomedical Research Centre. Microscopy was performed at the Light Microscopy Core Facility, UCL GOS Institute of Child Health, supported by the NIHR GOSH BRC award 17DD08. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, or the Department of Health. We thank Dr. Shyam Sawhney for technical support and the healthy volunteers who donated airway cells and blood samples for this study."