INTRODUCTION: Allergic asthmatics with both an early (EAR) and a late allergic reaction (LAR) following allergen exposure are termed 'dual responders' (DR), while 'single responders' (SR) only have an EAR. Mechanisms that differentiate DR from SR are largely unknown, particularly regarding the role and phenotypes of neutrophils. Therefore, we aimed to study neutrophils in DR and SR asthmatics. METHODS: Thirty-four allergic asthmatics underwent an inhaled allergen challenge, samples were collected before and up to 24 h post-challenge. Cell differentials were counted from bronchial lavage, alveolar lavage and blood; and tissue neutrophils were quantified in immune-stained bronchial biopsies. Lavage neutrophil nuclei lobe segmentation was used to classify active (1-4 lobes) from suppressive neutrophils (≥5 lobes). Levels of transmigration markers: soluble (s)CD62L and interleukin-1Ra, and activity markers: neutrophil elastase (NE), DNA-histone complex and dsDNA were measured in lavage fluid and plasma. RESULTS: Compared with SR at baseline, DR had more neutrophils in their bronchial airways at baseline, both in the lavage (p = .0031) and biopsies (p = .026) and elevated bronchial neutrophils correlated with less antitransmigratory IL-1Ra levels (r = -0.64). DR airways had less suppressive neutrophils and more 3-lobed (active) neutrophils (p = .029) that correlated with more bronchial lavage histone (p = .020) and more plasma NE (p = .0016). Post-challenge, DR released neutrophil extracellular trap factors in the blood earlier and had less pro-transmigratory sCD62L during the late phase (p = .0076) than in SR. CONCLUSION: DR have a more active airway neutrophil phenotype at baseline and a distinct neutrophil response to allergen challenge that may contribute to the development of an LAR. Therefore, neutrophil activity should be considered during targeted diagnosis and bio-therapeutic development for DR.

Center for Primary Health Care Research Department of Clinical Sciences Malmö Lund University Lund Sweden

Department of Clinical Sciences Lund Respiratory Medicine and Allergology Lund University Lund Sweden

Department of Experimental Medical Science Cell and Tissue biology Lund University Lund Sweden

Department of Microbiology Immunology and Transplantation KU Leuven Catholic University of Leuven Leuven Belgium

Department of Otorhinolaryngology Head and Neck Surgery Institute of Clinical Sciences Skane University Hospital Lund Sweden

Department of Respiratory Medicine 1st Faculty of Medicine Charles University and Thomayer Hospital Prague Czech Republic

Division of ENT Diseases Department of Clinical Science Intervention and Technology Karolinska Institutet Stockholm Sweden

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Diamant Z, Gauvreau GM, Cockcroft DW, et al. Inhaled allergen bronchoprovocation tests. J Allergy Clin Immunol. 2013;132(5):1045‐55.e6. PubMed

Machado L, Stalenheim G, Malmberg P. Early and late allergic bronchial reactions: physiological characteristics. Clin Allergy. 1986;16(2):111‐117. PubMed

Avila PC, Segal MR, Wong HH, Boushey HA, Fahy JV. Predictors of late asthmatic response. Logistic regression and classification tree analyses. Am J Respiratory Crit Med. 2000;161(6):2092‐2095. PubMed

Singh A, Shannon CP, Kim YW, et al. Novel blood‐based transcriptional biomarker panels predict the late‐phase asthmatic response. Am J Respir Crit Care Med. 2018;197(4):450‐462. PubMed

Singh A, Cohen Freue GV, Oosthuizen JL, et al. Plasma proteomics can discriminate isolated early from dual responses in asthmatic individuals undergoing an allergen inhalation challenge. Proteom – Clin Appl. 2012;6(9–10):476‐485. PubMed

Singh A, Yamamoto M, Kam SHY, et al. Gene‐metabolite expression in blood can discriminate allergen‐induced isolated early from dual asthmatic responses. PLoS One. 2013;8(7):e67907. PubMed PMC

Snelgrove RJ, Patel DF, Patel T, Lloyd CM. The enigmatic role of the neutrophil in asthma: friend, foe or indifferent? Clin Exp Allergy. 2018;48(10):1275‐1285. PubMed

Marwick JA, Mills R, Kay O, et al. Neutrophils induce macrophage anti‐inflammatory reprogramming by suppressing NF‐κB activation. Cell Death Dis. 2018;9(6):665. PubMed PMC

Leitch AE, Duffin R, Haslett C, Rossi AG. Relevance of granulocyte apoptosis to resolution of inflammation at the respiratory mucosa. Mucosal Immunol. 2008;1(5):350‐363. PubMed PMC

Moore WC, Hastie AT, Li X, et al. Sputum neutrophil counts are associated with more severe asthma phenotypes using cluster analysis. J Allergy Clin Immunol. 2014;133(6):1557‐63.e5. PubMed PMC

Just J, Gouvis‐Echraghi R, Rouve S, Wanin S, Moreau D, Annesi‐Maesano I. Two novel, severe asthma phenotypes identified during childhood using a clustering approach. Eur Respir J. 2012;40(1):55. PubMed

Nguyen LT, Lim S, Oates T, Chung KF. Increase in airway neutrophils after oral but not inhaled corticosteroid therapy in mild asthma. Respir Med. 2005;99(2):200‐207. PubMed

Radermecker C, Louis R, Bureau F, Marichal T. Role of neutrophils in allergic asthma. Curr Opin Immunol. 2018;54:28‐34. PubMed

Jin W, Dong C. IL‐17 cytokines in immunity and inflammation. Emerg. Microbes Infect. 2013;2(9):e60. PubMed PMC

Moore BB, Kunkel SL. Attracting attention: discovery of IL‐8/CXCL8 and the birth of the chemokine field. J Immunol (Baltimore, Md: 1950). 2019;202(1):3‐4. PubMed PMC

Hernandez ML, Mills K, Almond M, et al. IL‐1 receptor antagonist reduces endotoxin‐induced airway inflammation in healthy volunteers. J Allergy Clin Immunol. 2015;135(2):379‐385. PubMed PMC

Kolaczkowska E, Kubes P. Neutrophil recruitment and function in health and inflammation. Nat Rev Immunol. 2013;13(3):159‐175. PubMed

Ivetic A. A head‐to‐tail view of L‐selectin and its impact on neutrophil behaviour. Cell Tissue Res. 2018;371(3):437‐453. PubMed PMC

Toussaint M, Jackson DJ, Swieboda D, et al. Host DNA released by NETosis promotes rhinovirus‐induced type‐2 allergic asthma exacerbation. Nat Med. 2017;23(6):681‐691. PubMed PMC

Mortaz E, Alipoor SD, Adcock IM, Mumby S, Koenderman L. Update on neutrophil function in severe inflammation. Front Immunol. 2018;9:2171. PubMed PMC

Pillay J, Kamp VM, van Hoffen E, et al. A subset of neutrophils in human systemic inflammation inhibits T cell responses through Mac‐1. J Clin Invest. 2012;122(1):327‐336. PubMed PMC

Tak T, Wijten P, Heeres M, et al. Human CD62L(dim) neutrophils identified as a separate subset by proteome profiling and in vivo pulse‐chase labeling. Blood. 2017;129(26):3476‐3485. PubMed

Pillay J, Ramakers BP, Kamp VM, et al. Functional heterogeneity and differential priming of circulating neutrophils in human experimental endotoxemia. J Leukoc Biol. 2010;88(1):211‐220. PubMed

Ekstedt S, Stenberg H, Tufvesson E, et al. The potential role of CD16 high CD62L dim neutrophils in the allergic asthma. Allergy. 2019;74(11):2265‐2268. PubMed

Monteseirin J, Chacon P, Vega A, et al. L‐selectin expression on neutrophils from allergic patients. Clin Exp Allergy. 2005;35(9):1204‐1213. PubMed

Stenberg H, Diamant Z, Ankerst J, Bjermer L, Tufvesson E. Small airway involvement in the late allergic response in asthma. Clin Exp Allergy. 2017;47(12):1555‐1565. PubMed

Stenberg H, Wadelius E, Moitra S, et al. Club cell protein (CC16) in plasma, bronchial brushes, BAL and urine following an inhaled allergen challenge in allergic asthmatics. Biomarkers. 2018;23(1):51‐60. PubMed

Van Vyve T, Chanez P, Lacoste JY, Bousquet J, Michel FB, Godard P. Comparison between bronchial and alveolar samples of bronchoalveolar lavage fluid in asthma. Chest. 1992;102(2):356‐361. PubMed

Maretta M, Toth S, Jonecova Z, et al. Immunohistochemical expression of MPO, CD163 and VEGF in inflammatory cells in acute respiratory distress syndrome: a case report. Int J Clin Exp Pathol. 2014;7(7):4539‐4544. PubMed PMC

Sharshar RS. Impulse oscillometry in Small airway dysfunction in asthmatics and its utility in asthma control. Eur Respir J. 2018;52(suppl 62):PA1700.

Lommatzsch M, Julius P, Kuepper M, et al. The course of allergen‐induced leukocyte infiltration in human and experimental asthma. J Allergy Clin Immunol. 2006;118(1):91‐97. PubMed

Rahman I, Collado Sánchez A, Davies J, et al. L‐selectin regulates human neutrophil transendothelial migration. J Cell Sci. 2021;134(3):jcs250340. PubMed PMC

Zhang J, Bai C. Elevated serum interleukin‐8 level as a preferable biomarker for identifying uncontrolled asthma and glucocorticosteroid responsiveness. Tanaffos. 2017;16(4):260‐269. PubMed PMC

Ghaffari J, Rafiei AR, Ajami A, Mahdavi M, Hoshiar B. Serum interleukins 6 and 8 in mild and severe asthmatic patients, is it difference? Caspian J Intern Med. 2011;2(2):226‐228. PubMed PMC

Jappe U, Schwager C, Schromm AB, et al. Lipophilic allergens, different modes of allergen‐lipid interaction and their impact on asthma and allergy. Front Immunol. 2019;10:122. PubMed PMC

Wallin A, Pourazar J, Sandström T. LPS‐induced bronchoalveolar neutrophilia; effects of salmeterol treatment. Respir Med. 2004;98(11):1087‐1092. PubMed

Uddin M, Watz H, Malmgren A, Pedersen F. NETopathic inflammation in chronic obstructive pulmonary disease and severe asthma. Front Immunol. 2019;10:47. PubMed PMC

Wright TK, Gibson PG, Simpson JL, McDonald VM, Wood LG, Baines KJ. Neutrophil extracellular traps are associated with inflammation in chronic airway disease. Respirology. 2016;21(3):467‐475. PubMed

Agache I, Akdis CA, Akdis M, et al. EAACI biologicals guidelines – Recommendations for severe asthma. Allergy. 2020;76(1):14‐44. PubMed

Kyriakopoulos C, Gogali A, Bartziokas K, Kostikas K. Identification and treatment of T2‐low asthma in the era of biologics. ERJ Open Res. 2021;7(2):00309‐2020. PubMed PMC

Maas SL, Soehnlein O, Viola JR. Organ‐specific mechanisms of transendothelial neutrophil migration in the lung, liver, kidney, and aorta. Front Immunol. 2018;9:2739. PubMed PMC

Gao P, Gibson PG, Baines KJ, et al. Anti‐inflammatory deficiencies in neutrophilic asthma: reduced galectin‐3 and IL‐1RA/IL‐1β. Respir Res. 2015;16(1):5. PubMed PMC

Evans MD, Esnault S, Denlinger LC, Jarjour NN. Sputum cell IL‐1 receptor expression level is a marker of airway neutrophilia and airflow obstruction in asthmatic patients. J Allergy Clin Immunol. 2018;142(2):415‐423. PubMed PMC

Ronchetti S, Ricci E, Migliorati G, Gentili M, Riccardi C. How glucocorticoids affect the neutrophil life. Int J Mol Sci. 2018;19(12):4090. PubMed PMC

Baatjes AJ, Smith SG, Watson R, et al. T regulatory cell phenotypes in peripheral blood and bronchoalveolar lavage from non‐asthmatic and asthmatic subjects. Clin Exp Allergy. 2015;45(11):1654‐1662. PubMed

Becker N, Störmann P, Janicova A, et al. Club cell protein 16 attenuates CD16(bright)CD62(dim) immunosuppressive neutrophils in damaged tissue upon posttraumatic sepsis‐induced lung injury. J Immunol Res. 2021;2021:6647753. PubMed PMC

Donnelly SC, Haslett C, Robertson CE, et al. Role of selectiris in development of adult respiratory distress syndrome. The Lancet. 1994;344(8917):215‐219. PubMed

Strausbaugh HJ, Green PG, Lo E, et al. Painful stimulation suppresses joint inflammation by inducing shedding of L‐selectin from neutrophils. Nat Med. 1999;5(9):1057‐1061. PubMed

Prame Kumar K, Nicholls AJ, Wong CHY. Partners in crime: neutrophils and monocytes/macrophages in inflammation and disease. Cell Tissue Res. 2018;371(3):551‐565. PubMed PMC

Eguíluz‐Gracia I, Malmstrom K, Dheyauldeen SA, et al. Monocytes accumulate in the airways of children with fatal asthma. Clin Exp Allergy. 2018;48(12):1631‐1639. PubMed