High expression of neutrophil and monocyte CD64 with simultaneous lack of upregulation of adhesion receptors CD11b, CD162, CD15, CD65 on neutrophils in severe COVID-19

Malgorzata Karawajczyk, Lena Douhan Håkansson, Miklos Lipcsey, Michael Hultström, Karlis Pauksens, Robert Frithiof, Anders Larsson, Malgorzata Karawajczyk, Lena Douhan Håkansson, Miklos Lipcsey, Michael Hultström, Karlis Pauksens, Robert Frithiof, Anders Larsson

Abstract

Background and aims: The pronounced neutrophilia observed in patients with coronavirus disease 2019 (COVID-19) infections suggests a role for these leukocytes in the pathology of the disease. Monocyte and neutrophil expression of CD64 and CD11b have been reported as early biomarkers to detect infections. The aim of this study was to study the expression of receptors for IgG (CD64) and adhesion molecules (CD11b, CD15s, CD65, CD162, CD66b) on neutrophils and monocytes in patients with severe COVID-19 after admission to an intensive care unit (ICU).

Methods: The expression of receptors was analyzed using flow cytometry. EDTA blood from 23 patients with confirmed COVID-19 infection was sampled within 48 h of admission to the ICU. Leukocytes were labeled with antibodies to CD11b, CD15s, CD65s, CD162, CD64, and CD66b. Expression of receptors was reported as mean fluorescence intensity (MFI) or the percentage of cells expressing receptors.

Results: Results are presented as comparison of COVID-19 patients with the healthy group and the receptor expression as MFI. Neutrophil receptors CD64 (2.5 versus 0.5) and CD66b (44.5 versus 34) were increased and CD15 decreased (21.6 versus 28.3) when CD65 (6.6 versus 4.4), CD162 (21.3 versus 21.1) and CD11b (10.5 versus 12) were in the same range. Monocytes receptors CD64 (30.5 versus 16.6), CD11b (18.7 versus 9.8), and CD162 (38.6 versus 36.5) were increased and CD15 decreased (10.3 versus 17.9); CD65 were in the same range (2.3 versus 1.96).

Conclusion: Monocytes and neutrophils are activated during severe COVID-19 infection as shown by strong upregulation of CD64. High monocyte and neutrophil CD64 can be an indicator of a severe form of COVID19. The adhesion molecules (CD11b, CD162, CD65, and CD15) are not upregulated on otherwise activated neutrophils, which might lead to relative impairment of tissue migration. Low adhesion profile of neutrophils suggests immune dysfunction of neutrophils. Monocytes maintain upregulation of some adhesion molecules (CD11b, CD162) suggesting the persistence of an increased ability to migrate into tissues, even during a severe stage of COVID-19. Future research should focus on CD64 and CD11b kinetics in the context of prognosis.

Keywords: CD 162; CD11b; CD15; CD64; CD65; COVID-19; monocyte; neutrophil.

Conflict of interest statement

Conflict of interest statement: The authors declare that there is no conflict of interest.

© The Author(s), 2021.

Figures

Figure 1.
Figure 1.
(a) Expression of CD11b and CD15s on monocytes and neutrophils. Bars represent median values. A difference is considered significant if p < 0.05 as calculated with the Benjamin–Hochberg test and p < 0.01 with the Mann–Whitney test. (•) CD11b monocytes, (■) CD11b neutrophils, (○) CD15s monocytes, (□) CD15s neutrophil. (b) Expression of CD65 and CD162 on monocytes and neutrophils. Bars represent the mean value. A difference is considered significant if p < 0.05 as calculated with the Benjamin–Hochberg test and p < 0.01 with the Mann–Whitney test. (•) CD65 monocytes, (■) CD65 neutrophils, (○) CD162 monocytes, (□) CD162 neutrophils. COVID-19, coronavirus disease 2019; MFI, mean fluorescence intensity.
Figure 2.
Figure 2.
Expression of CD64 on monocytes and neutrophils. Bars represent the mean value. A difference is considered significant if p < 0.05 as calculated with the Benjamin–Hochberg test and p < 0.01 with the Mann–Whitney test. (•) CD64 monocytes MFI, (♦) CD64 neutrophils MFI, (□) CD64 neutrophils %. COVID-19, coronavirus disease 2019; MFI, mean fluorescence intensity.
Figure 3.
Figure 3.
ROC curves for predicting death. Dotted lines show ROC curve, light grey CI (a) monocyte CD15 AUC = 0.82 (b) monocyte CD64 AUC = 0.79, (c) troponin AUC = 0.86 and (d) NT-pro-BNP AUC = 0.833. AUC, area under the ROC curve; CI, confidence interval; COVID-19, coronavirus disease 2019; MFI, mean fluorescence intensity; ROC, receiver operating characteristic.

References

    1. Liu J, Li H, Luo M, et al.. Lymphopenia predicted illness severity and recovery in patients with COVID-19: a single-center, retrospective study. PLoS One 2020; 15: e0241659.
    1. Liu J, Liu Y, Xiang P, et al.. Neutrophil-to-lymphocyte ratio predicts critical illness patients with 2019 coronavirus disease in the early stage. J Transl Med 2020; 18: 206.
    1. Mortaz E, Alipoor SD, Adcock IM, et al.. Update on neutrophil function in severe inflammation. Front Immunol 2018; 9: 2171.
    1. Barnes BJ, Adrover JM, Baxter-Stoltzfus A, et al.. Targeting potential drivers of COVID-19: neutrophil extracellular traps. J Exp Med. Epub ahead of print 1 June 2020. DOI: 10.1084/jem.20200652.
    1. Bülow Anderberg S, Luther T, Berglund M, et al.. Increased levels of plasma cytokines and correlations to organ failure and 30-day mortality in critically ill Covid-19 patients. Cytokine 2021; 138: 155389.
    1. Vitte J, Diallo AB, Boumaza A, et al.. A granulocytic signature identifies COVID-19 and its severity. J Infect Dis 2020; 222: 1985–1996.
    1. Rawson TM, Moore LSP, Zhu N, et al.. Bacterial and fungal coinfection in individuals with coronavirus: a rapid review to support COVID-19 antimicrobial prescribing. Clin Infect Dis 2020; 71: 2459–2468.
    1. Shallcross LJ, Davies DSC. Antibiotic overuse: a key driver of antimicrobial resistance. Br J Gen Pract 2014; 64: 604–605.
    1. Cassini A, Högberg LD, Plachouras D, et al.. Attributable deaths and disability-adjusted life-years caused by infections with antibiotic-resistant bacteria in the EU and the European Economic Area in 2015: a population-level modelling analysis. Lancet Infect Dis 2019; 19: 56–66.
    1. Spalding MC, Cripps MW, Minshall CT. Ventilator-associated pneumonia: new definitions. Crit Care Clin 2017; 33: 277–292.
    1. Ioan-Facsinay A, de Kimpe SJ, Hellwig SMM, et al.. FcγRI (CD64) contributes substantially to severity of arthritis, hypersensitivity responses, and protection from bacterial infection. Immunity 2002; 16: 391–402.
    1. Borregaard N, Lollike K, Kjeldsen L, et al.. Human neutrophil granules and secretory vesicles. Eur J Haematol 1993; 51: 187–198.
    1. Cid J, Aguinaco R, Sánchez R, et al.. Neutrophil CD64 expression as marker of bacterial infection: a systematic review and meta-analysis. J Infect 2010; 60: 313–319.
    1. Hashem HE, Abdel Halim RM, El Masry SA, et al.. The utility of neutrophil CD64 and presepsin as diagnostic, prognostic, and monitoring biomarkers in neonatal sepsis. Int J Microbiol 2020; 2020: 8814892.
    1. Lu L, Jin X, Zhang Q. CD35 and CD64 of neutrophils can differentiate between bacterial and viral infections in children by simultaneous quantitative analysis. Med Sci Monit 2019; 25: 7728–7734.
    1. Fjaertoft G, Pauksen K, Håkansson L, et al.. Cell surface expression of FcgammaRI (CD64) on neutrophils and monocytes in patients with influenza A, with and without complications. Scand J Infect Dis 2005; 37: 882–889.
    1. Jämsä J, Huotari V, Savolainen E-R, et al.. Kinetics of leukocyte CD11b and CD64 expression in severe sepsis and non-infectious critical care patients. Acta Anaesthesiol Scand 2015; 59: 881–891.
    1. Hu B-Q, Yang Y, Zhao C-J, et al.. Accuracy of neutrophil CD64 expression in diagnosing infection in patients with autoimmune diseases: a meta-analysis. Clin Rheumatol 2019; 38: 1319–1328.
    1. Muzlovic I, Ihan A, Stubljar D. CD64 index on neutrophils can diagnose sepsis and predict 30-day survival in subjects after ventilator-associated pneumonia. J Infect Dev Ctries 2016; 10: 260–268.
    1. Jämsä J, Ala-Kokko T, Huotari V, et al.. Neutrophil CD64, C-reactive protein, and procalcitonin in the identification of sepsis in the ICU - post-test probabilities. J Crit Care 2018; 43: 139–142.
    1. McEver RP. Selectins: initiators of leucocyte adhesion and signalling at the vascular wall. Cardiovasc Res 2015; 107: 331–339.
    1. Pauksens K, Fjaertoft G, Douhan-Håkansson L, et al.. Neutrophil and monocyte receptor expression in uncomplicated and complicated influenza A infection with pneumonia. Scand J Infect Dis 2008; 40: 326–337.
    1. Qiu X, Li J, Yang X, et al.. Is neutrophil CD11b a special marker for the early diagnosis of sepsis in neonates? A systematic review and meta-analysis. BMJ Open. Epub ahead of print 1 May 2019. DOI: 10.1136/bmjopen-2018-025222.
    1. Zhou H, Li Y, Gui H, et al.. Antagonism of integrin CD11b affords protection against endotoxin shock and polymicrobial sepsis via attenuation of HMGB1 nucleocytoplasmic translocation and extracellular release. J Immunol 2018; 200: 1771–1780.
    1. Zhang S, Song L, Wang Y, et al.. Targeting CD162 protects against streptococcal M1 protein-evoked neutrophil recruitment and lung injury. Am J Physiol Lung Cell Mol Physiol 2013; 305: L756–L763.
    1. Mallia P, Message SD, Contoli M, et al.. Neutrophil adhesion molecules in experimental rhinovirus infection in COPD. Respir Res 2013; 14: 72.
    1. Stålhammar ME, Sindelar R, Douhan Håkansson L. Neutrophil receptor response to bacterial N-formyl peptides is similar in term newborn infants and adults in contrast to IL-8. Scand J Immunol 2016; 84: 332–337.
    1. Noguchi M, Sato N, Sugimori H, et al.. A minor E-selectin ligand, CD65, is critical for extravascular infiltration of acute myeloid leukemia cells. Leuk Res 2001; 25: 847–853.
    1. Hammarström S. The carcinoembryonic antigen (CEA) family: structures, suggested functions and expression in normal and malignant tissues. Semin Cancer Biol 1999; 9: 67–81.
    1. Torsteinsdóttir I, Arvidson NG, Hällgren R, et al.. Enhanced expression of integrins and CD66b on peripheral blood neutrophils and eosinophils in patients with rheumatoid arthritis, and the effect of glucocorticoids. Scand J Immunol 1999; 50: 433–439.
    1. Zhao L, Xu S, Fjaertoft G, et al.. An enzyme-linked immunosorbent assay for human carcinoembryonic antigen-related cell adhesion molecule 8, a biological marker of granulocyte activities in vivo. J Immunol Methods 2004; 293: 207–214.
    1. Hamblin A, Taylor M, Bernhagen J, et al.. A method of preparing blood leucocytes for flow cytometry which prevents upregulation of leucocyte integrins. J Immunol Methods 1992; 146: 219–228.
    1. Moreno RP, Metnitz PGH, Almeida E, et al.. SAPS 3–from evaluation of the patient to evaluation of the intensive care unit. Part 2: development of a prognostic model for hospital mortality at ICU admission. Intensive Care Med 2005; 31: 1345–1355.
    1. Simms H, D’Amico R. Regulation of polymorphonuclear neutrophil CD16 and CD11b/CD18 expression by matrix proteins during hypoxia is VLA-5, VLA-6 dependent. J Immunol Baltim Md 1950 1995; 155: 4979–4990.
    1. Leino L, Lilius EM. The up- and down-modulation of immunoglobulin G Fc receptors and complement receptors on activated human neutrophils depends on the nature of activator. J Leukoc Biol 1992; 51: 157–163.
    1. Carter MJ, Fish M, Jennings A, et al.. Peripheral immunophenotypes in children with multisystem inflammatory syndrome associated with SARS-CoV-2 infection. Nat Med 2020; 26: 1701–1707.
    1. Hashem HE, El Masry SA, Mokhtar AM, et al.. Valuable role of neutrophil CD64 and highly sensitive CRP biomarkers for diagnostic, monitoring, and prognostic evaluations of sepsis patients in neonatal ICUs. BioMed Res Int 2020; 2020: 6214363.
    1. Qin J-J, Cheng X, Zhou F, et al.. Redefining cardiac biomarkers in predicting mortality of inpatients with COVID-19. Hypertension 2020; 76: 1104–1112.
    1. Malik P, Patel U, Mehta D, et al.. Biomarkers and outcomes of COVID-19 hospitalisations: systematic review and meta-analysis. BMJ Evid-Based Med. Epub ahead of print 15 September 2020. DOI: 10.1136/bmjebm-2020-111536.
    1. Krutmann J, Kirnbauer R, Köck A, et al.. Cross-linking Fc receptors on monocytes triggers IL-6 production. Role in anti-CD3-induced T cell activation. J Immunol Baltim Md 1950 1990; 145: 1337–1342.
    1. Tanaka M, Krutzik SR, Sieling PA, et al.. Activation of Fc gamma RI on monocytes triggers differentiation into immature dendritic cells that induce autoreactive T cell responses. J Immunol 2009; 183: 2349–2355.
    1. Meizlish ML, Pine AB, Bishai JD, et al.. A neutrophil activation signature predicts critical illness and mortality in COVID-19. MedRxiv. Epub ahead of print 2 September 2020. DOI: 10.1101/2020.09.01.20183897.
    1. Nicolai L, Leunig A, Brambs S, et al.. Vascular neutrophilic inflammation and immunothrombosis distinguish severe COVID-19 from influenza pneumonia. J Thromb Haemost. Epub ahead of print 20 November 2020. DOI: 10.1111/jth.15179.

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