Your browser doesn't support javascript.
Show: 20 | 50 | 100
Results 1 - 20 de 765
Filter
1.
Ann Afr Med ; 21(4): 371-376, 2022.
Article in English | MEDLINE | ID: covidwho-2144095

ABSTRACT

Introduction: COVID 19 pandemic has given rise to several challenges to clinicians and one of the keys in this is to predict the set of patients who progress from mild disease to moderate and severe. Apart from the symptomatology and signs, there are several lab parameters varying from biochemical, hematological to radiological parameters which help us in stratifying the stage of disease and also in deciding on which set of patients need close and vigilant monitoring. This would help us in better stratification of disease and utilize the available infrastructure and resources in an optimum way for better management of the disease. Aim: To analyze the early warning efficiency of laboratory parameters individually or in combination in predicting the progress of disease in patients from mild to moderate/severe disease. Materials and Methods: This was taken up as a retrospective study with 100 cases and 100 controls. The demographic details, inflammatory markers, biochemical markers and hematological markers were analyzed. Test of significance was employed to compare categorical variables while student t-test was employed to test the difference in the mean value such as age between case and control (Mann-Whitney U-test in parameters not having normal distribution). Receiver operating characteristic (ROC) curve was constructed for these parameters using cases and controls and area under the curve (AUC) were estimated which was used as an indicator of sensitivity and specificity of the parameter in their early warning efficiency. The critical values for each of the parameters either individually or in combination was estimated as well. Results: Among the parameters C reactive protein (CRP), d-dimers and eosinopenia have the best early warning efficiency. The area under the ROCs curve for neutrophil lymphocyte ratio (NLR), CRP. Ferritin, lactate dehydrogenase, Eosinopenia was 0.609, 0.947, 0.614, 0.554, 0.617 respectively at triage. However, a combination of eosinopenia with CRP (AUC-0.732) or NLR with CRP (AUC-0.728) have a good sensitivity and specificity in predicting the outcome regarding the progression of the disease. Conclusions: Among the parameters, CRP, d-dimers, Eosinopenia and NLR have the best early warning efficiency. However, a combination of Eosinopenia and CRP at triage should also serve as a red flag sign in patients apart from the well-known NLR and IL6 values.


Résumé Introduction: La pandémie covide 19 a relevé plusieurs défis aux cliniciens et l'une des clés dans ce domaine est de prédire l'ensemble des patients qui passent d'une maladie légère à modérée et sévère. Outre la symptomatologie et les signes, plusieurs paramètres de laboratoire variant des paramètres biochimiques, hématologiques à radiologiques qui nous aident à stratifier le stade de la maladie et également à décider quel ensemble de patients nécessite une surveillance étroite et vigilante. Cela nous aiderait à mieux stratification des maladies et à utiliser l'infrastructure et les ressources disponibles de manière optimale pour une meilleure prise en charge de la maladie. Objectif: Analyser l'efficacité d'alerte précoce des paramètres de laboratoire individuellement ou en combinaison pour prédire les progrès des maladies chez les patients d'une maladie légère à modérée / sévère. Matériaux et méthodes: Ceci a été considéré comme une étude rétrospective avec 100 cas et 100 contrôles. Les détails démographiques, les marqueurs inflammatoires, les marqueurs biochimiques et les marqueurs hématologiques ont été analysés. Le test de signification a été utilisé pour comparer les variables catégorielles tandis que le test T des étudiants a été utilisé pour tester la différence de valeur moyenne telle que l'âge entre le cas et le contrôle (test U Mann - Whitney dans les paramètres n'ayant pas de distribution normale). La courbe des caractéristiques de fonctionnement du récepteur (ROC) a été construite pour ces paramètres en utilisant les cas et les contrôles et la zone sous la courbe (AUC) ont été estimés qui ont été utilisés comme indicateur de sensibilité et de spécificité du paramètre dans leur efficacité d'alerte précoce. Les valeurs critiques pour chacun des paramètres individuellement ou en combinaison ont également été estimées. Résultats: Parmi les paramètres C Protein réactif (CRP), les D - dimères et l'éosinopénie ont la meilleure efficacité d'alerte précoce. La zone sous la courbe ROCS pour le rapport lymphocyte des neutrophiles (NLR), CRP. La ferritine, la lactate déshydrogénase, l'éosinopénie était de 0,609, 0,947, 0,614, 0,554, 0,617 respectivement au triage. Cependant, une combinaison d'éosinopénie avec CRP (AUC - 0,732) ou NLR avec CRP (AUC - 0,728) a une bonne sensibilité et spécificité pour prédire le résultat concernant la progression de la maladie. Conclusions: Parmi les paramètres, le CRP, les D - dimères, l'éosinopénie et le NLR ont la meilleure efficacité d'alerte précoce. Cependant, une combinaison d'éosinopénie et de CRP au triage devrait également servir de signe du drapeau rouge chez les patients en dehors des valeurs NLR et IL6 bien connues. Mots-clés: C Protéine réactive, efficacité d'alerte précoce, éosinopénie, progression de la maladie dans Covid ­ 19.


Subject(s)
COVID-19 , Humans , Retrospective Studies , Lymphocytes , Neutrophils , Biomarkers
2.
Cells ; 11(22)2022 Nov 16.
Article in English | MEDLINE | ID: covidwho-2142561

ABSTRACT

Alveolar macrophage (AM) proliferation and self-renewal play an important role in the lung tissue microenvironment. However, the impact of immune cells, especially the neutrophils, on AM homeostasis or function is not well characterized. In this study, we induced in vivo migration of neutrophils into bronchoalveolar lavage (BAL) fluid and lung using CXCL1, and then co-cultured these with AMs in vitro. Neutrophils in the BAL (BAL-neutrophils), rather than neutrophils of bone marrow (BM-neutrophils), were found to inhibit AM proliferation. Analysis of publicly available data showed high heterogeneity of lung neutrophils with distinct molecular signatures of BM- and blood-neutrophils. Unexpectedly, BAL-neutrophils from influenza virus PR8-infected mice (PR8-neutrophils) did not inhibit the proliferation of AMs. Bulk RNA sequencing further revealed that co-culture of AMs with PR8-neutrophils induced IFN-α and -γ responses and inflammatory response, and AMs co-cultured with BAL-neutrophils showed higher expression of metabolism- and ROS-associated genes; in addition, BAL-neutrophils from PR8-infected mice modulated AM polarization and phagocytosis. BAL-neutrophil-mediated suppression of AM proliferation was abrogated by a combination of inhibitors of different neutrophil death pathways. Collectively, our findings suggest that multiple cell death pathways of neutrophils regulate the proliferation of AMs. Targeting neutrophil death may represent a potential therapeutic strategy for improving AM homeostasis during respiratory diseases.


Subject(s)
Macrophages, Alveolar , Neutrophils , Mice , Animals , Macrophages, Alveolar/metabolism , Neutrophils/metabolism , Bronchoalveolar Lavage Fluid , Lung , Cell Proliferation
3.
Front Immunol ; 13: 993720, 2022.
Article in English | MEDLINE | ID: covidwho-2142018

ABSTRACT

Pathogenesis of lung injury in COVID-19 is not completely understood, leaving gaps in understanding how current treatments modulate the course of COVID-19. Neutrophil numbers and activation state in circulation have been found to correlate with COVID-19 severity, and neutrophil extracellular traps (NETs) have been found in the lung parenchyma of patients with acute respiratory distress syndrome (ARDS) in COVID-19. Targeting the pro-inflammatory functions of neutrophils may diminish lung injury in COVID-19 and ARDS. Neutrophils were isolated from peripheral blood of healthy donors, treated ex vivo with dexamethasone, tocilizumab and intravenous immunoglobulin (IVIG) and NET formation, oxidative burst, and phagocytosis were assessed. Plasma from critically ill COVID-19 patients before and after clinical treatment with IVIG and from healthy donors was assessed for neutrophil activation-related proteins. While dexamethasone and tocilizumab did not affect PMA- and nigericin-induced NET production ex vivo, IVIG induced a dose-dependent abrogation of NET production in both activation models. IVIG also reduced PMA-elicited reactive oxygen species production, but did not alter phagocytosis. COVID-19 patients were found to have elevated levels of cell-free DNA, neutrophil elastase and IL-8 as compared to healthy controls. Levels of both cell-free DNA and neutrophil elastase were lower 5 days after 4 days of daily treatment with IVIG. The lack of impact of dexamethasone or tocilizumab on these neutrophil functions suggests that these therapeutic agents may not act through suppression of neutrophil functions, indicating that the door might still be open for the addition of a neutrophil modulator to the COVID-19 therapeutic repertoire.


Subject(s)
COVID-19 , Cell-Free Nucleic Acids , Lung Injury , Respiratory Distress Syndrome , Humans , Neutrophils/metabolism , Immunoglobulins, Intravenous/therapeutic use , Immunoglobulins, Intravenous/pharmacology , COVID-19/drug therapy , Leukocyte Elastase/metabolism , Lung Injury/metabolism , Cell-Free Nucleic Acids/metabolism , Dexamethasone
4.
Front Immunol ; 13: 931388, 2022.
Article in English | MEDLINE | ID: covidwho-2141951

ABSTRACT

Intracranial inoculation of the neuroadapted JHM strain of mouse hepatitis virus (JHMV) into susceptible strains of mice results in acute encephalomyelitis followed by a cimmune-mediated demyelination similar to the human demyelinating disease multiple sclerosis (MS). JHMV infection of transgenic mice in which expression of the neutrophil chemoattractant chemokine CXCL1 is under the control of a tetracycline-inducible promoter active within GFAP-positive cells results in sustained neutrophil infiltration in the central nervous system (CNS) that correlates with an increase in spinal cord demyelination. We used single cell RNA sequencing (scRNAseq) and flow cytometry to characterize molecular and cellular changes within the CNS associated with increased demyelination in transgenic mice compared to control animals. These approaches revealed the presence of activated neutrophils as determined by expression of mRNA transcripts associated with neutrophil effector functions, including CD63, MMP9, S100a8, S100a9, and ASPRV1, as well as altered neutrophil morphology and protein expression. Collectively, these findings reveal insight into changes in the profile of neutrophils associated with increased white matter damage in mice persistently infected with a neurotropic coronavirus.


Subject(s)
Demyelinating Diseases , Multiple Sclerosis , Murine hepatitis virus , White Matter , Animals , Central Nervous System , Chemokine CXCL1/metabolism , Humans , Matrix Metalloproteinase 9/metabolism , Mice , Mice, Transgenic , Multiple Sclerosis/metabolism , Neutrophils/metabolism , RNA, Messenger , Tetracyclines , White Matter/metabolism
5.
J Am Soc Nephrol ; 33(5): 936-947, 2022 05.
Article in English | MEDLINE | ID: covidwho-2141044

ABSTRACT

BACKGROUND: The ANCA autoantigens proteinase 3 (PR3) and myeloperoxidase (MPO) are exclusively expressed by neutrophils and monocytes. ANCA-mediated activation of these cells is the key driver of the vascular injury process in ANCA-associated vasculitis (AAV), and neutrophil serine proteases (NSPs) are disease mediators. Cathepsin C (CatC) from zymogens activates the proteolytic function of NSPs, including PR3. Lack of NSP zymogen activation results in neutrophils with strongly reduced NSP proteins. METHODS: To explore AAV-relevant consequences of blocking NSP zymogen activation by CatC, we used myeloid cells from patients with Papillon-Lefèvre syndrome, a genetic deficiency of CatC, to assess NSPs and NSP-mediated endothelial cell injury. We also examined pharmacologic CatC inhibition in neutrophil-differentiated human hematopoietic stem cells, primary human umbilical vein cells, and primary glomerular microvascular endothelial cells. RESULTS: Patients with Papillon-Lefèvre syndrome showed strongly reduced NSPs in neutrophils and monocytes. Neutrophils from these patients produced a negative PR3-ANCA test, presented less PR3 on the surface of viable and apoptotic cells, and caused significantly less damage in human umbilical vein cells. These findings were recapitulated in human stem cells, in which a highly specific CatC inhibitor, but not prednisolone, reduced NSPs without affecting neutrophil differentiation, reduced membrane PR3, and diminished neutrophil activation upon PR3-ANCA but not MPO-ANCA stimulation. Compared with healthy controls, neutrophils from patients with Papillon-Lefèvre syndrome transferred less proteolytically active NSPs to glomerular microvascular endothelial cells, the cell type targeted in ANCA-induced necrotizing crescentic glomerulonephritis. Finally, both genetic CatC deficiency and pharmacologic inhibition, but not prednisolone, reduced neutrophil-induced glomerular microvascular endothelial cell damage. CONCLUSIONS: These findings may offer encouragement for clinical studies of adjunctive CatC inhibitor in patients with PR3-AAV.


Subject(s)
Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis , Papillon-Lefevre Disease , Antibodies, Antineutrophil Cytoplasmic , Cathepsin C/metabolism , Endothelial Cells/metabolism , Enzyme Precursors/metabolism , Humans , Myeloblastin/genetics , Neutrophils/metabolism , Papillon-Lefevre Disease/metabolism , Peroxidase
6.
Genome Med ; 14(1): 135, 2022 Nov 28.
Article in English | MEDLINE | ID: covidwho-2139390

ABSTRACT

BACKGROUND: As circulating DNA (cirDNA) is mainly detected as mononucleosome-associated circulating DNA (mono-N cirDNA) in blood, apoptosis has until now been considered as the main source of cirDNA. The mechanism of cirDNA release into the circulation, however, is still not fully understood. This work addresses that knowledge gap, working from the postulate that neutrophil extracellular traps (NET) may be a source of cirDNA, and by investigating whether NET may directly produce mono-N cirDNA. METHODS: We studied (1) the in vitro kinetics of cell derived genomic high molecular weight (gHMW) DNA degradation in serum; (2) the production of extracellular DNA and NET markers such as neutrophil elastase (NE) and myeloperoxidase (MPO) by ex vivo activated neutrophils; and (3) the in vitro NET degradation in serum; for this, we exploited the synergistic analytical information provided by specifically quantifying DNA by qPCR, and used shallow WGS and capillary electrophoresis to perform fragment size analysis. We also performed an in vivo study in knockout mice, and an in vitro study of gHMW DNA degradation, to elucidate the role of NE and MPO in effecting DNA degradation and fragmentation. We then compared the NET-associated markers and fragmentation size profiles of cirDNA in plasma obtained from patients with inflammatory diseases found to be associated with NET formation and high levels of cirDNA (COVID-19, N = 28; systemic lupus erythematosus, N = 10; metastatic colorectal cancer, N = 10; and from healthy individuals, N = 114). RESULTS: Our studies reveal that gHMW DNA degradation in serum results in the accumulation of mono-N DNA (81.3% of the remaining DNA following 24 h incubation in serum corresponded to mono-N DNA); "ex vivo" NET formation, as demonstrated by a concurrent 5-, 5-, and 35-fold increase of NE, MPO, and cell-free DNA (cfDNA) concentration in PMA-activated neutrophil culture supernatant, leads to the release of high molecular weight DNA that degrades down to mono-N in serum; NET mainly in the form of gHMW DNA generate mono-N cirDNA (2 and 41% of the remaining DNA after 2 h in serum corresponded to 1-10 kbp fragments and mono-N, respectively) independent of any cellular process when degraded in serum; NE and MPO may contribute synergistically to NET autocatabolism, resulting in a 25-fold decrease in total DNA concentration and a DNA fragment size profile similar to that observed from cirDNA following 8 h incubation with both NE and MPO; the cirDNA size profile of NE KO mice significantly differed from that of the WT, suggesting NE involvement in DNA degradation; and a significant increase in the levels of NE, MPO, and cirDNA was detected in plasma samples from lupus, COVID-19, and mCRC, showing a high correlation with these inflammatory diseases, while no correlation of NE and MPO with cirDNA was found in HI. CONCLUSIONS: Our work describes the mechanisms by which NET and cirDNA are linked. In doing so, we demonstrate that NET are a major source of mono-N cirDNA independent of apoptosis and establish a new paradigm of the mechanisms of cirDNA release in normal and pathological conditions. We also demonstrate a link between immune response and cirDNA.


Subject(s)
COVID-19 , Cell-Free Nucleic Acids , Extracellular Traps , Animals , Mice , Neutrophils , Genomics
7.
Front Immunol ; 13: 1065345, 2022.
Article in English | MEDLINE | ID: covidwho-2123419

ABSTRACT

Background: Several systemic inflammatory biomarkers have been associated with poor overall survival (OS) and disease severity in patients with coronavirus disease 2019 (COVID-19). However, it remains unclear which markers are better for predicting prognosis, especially for COVID-19 Omicron BA.2 infected patients. The present study aimed to identify reliable predictors of prognosis of COVID-19 Omicron BA.2 from inflammatory indicators. Methods: A cohort of 2645 COVID-19 Omicron BA.2 infected patients were retrospectively analyzed during the Omicron BA.2 surge in Shanghai between April 12, 2022, and June 17, 2022. The patients were admitted to the Shanghai Fourth People's Hospital, School of Medicine, Tongji University. Six systemic inflammatory indicators were included, and their cut-off points were calculated using maximally selected rank statistics. The analysis involved Kaplan-Meier curves, univariate and multivariate Cox proportional hazard models, and time-dependent receiver operating characteristic curves (time-ROC) for OS-associated inflammatory indicators. Results: A total of 2347 COVID-19 Omicron BA.2 infected patients were included. All selected indicators proved to be independent predictors of OS in the multivariate analysis (all P < 0.01). A high derived neutrophil to lymphocyte ratio (dNLR) was associated with a higher mortality risk of COVID-19 [hazard ratio, 4.272; 95% confidence interval (CI), 2.417-7.552]. The analyses of time-AUC and C-index showed that the dNLR (C-index: 0.844, 0.824, and 0.718 for the 5th, 10th, and 15th day, respectively) had the best predictive power for OS in COVID-19 Omicron BA.2 infected patients. Among different sub-groups, the dNLR was the best predictor for OS regardless of age (0.811 for patients aged ≥70 years), gender (C-index, 0.880 for men and 0.793 for women) and disease severity (C-index, 0.932 for non-severe patients and 0.658 for severe patients). However, the platelet to lymphocyte ratio was superior to the other indicators in patients aged <70 years. Conclusions: The prognostic ability of the dNLR was higher than the other evaluated inflammatory indicators for all COVID-19 Omicron BA.2 infected patients.


Subject(s)
COVID-19 , Neutrophils , Humans , Male , Female , Retrospective Studies , China/epidemiology , Lymphocytes , Prognosis
8.
J Infect Dev Ctries ; 16(10): 1564-1569, 2022 10 31.
Article in English | MEDLINE | ID: covidwho-2110321

ABSTRACT

INTRODUCTION: This study aims to research the effects of hematological and inflammatory parameters on the prognosis of COVID-19 disease and hospitalization duration. METHODOLOGY: One hundred and eighty-six patients with COVID-19 and a control group consisting of 187 healthy individuals were included in the study. Hematological variables and inflammatory parameters of the patients were recorded on the first and the fifth days of hospitalization. RESULTS: White blood cell count, lymphocyte count, and platelet count were statistically lower, and mean platelet volume (MPV), neutrophil to lymphocyte ratio (NLR), and platelet to lymphocyte ratio (PLR) levels were higher in the patient group compared to the control group. It was observed that the neutrophil count and MPV level were lower, and the platelet count and ferritin level were statistically higher on the fifth day of follow-up compared to the admission day. In contrast, there was a significantly positive correlation between the duration of hospitalization and the fifth day D-dimer (r = 0.546, p < 0.001) and ferritin (r = 0.568, p < 0.001); in addition, there was a negative correlation between the duration of hospitalization and admission day lymphocyte count and the fifth-day lymphocyte count. CONCLUSIONS: Increased levels of ferritin and D-dimer, and decreased count of lymphocytes are among the important factors affecting the duration of hospitalization for COVID-19 patients. Furthermore, we think that neutrophil count and MPV levels are low, and platelet count and ferritin levels are high during the disease. Therefore, these parameters can be used as prognostic indicators of the disease.


Subject(s)
COVID-19 , Humans , COVID-19/diagnosis , Retrospective Studies , Lymphocyte Count , Platelet Count , Leukocyte Count , Mean Platelet Volume , Lymphocytes , Neutrophils , Ferritins
9.
Front Immunol ; 13: 1010140, 2022.
Article in English | MEDLINE | ID: covidwho-2121437

ABSTRACT

The emerging SARS-CoV-2 virus has affected the entire world with over 600 million confirmed cases and 6.5 million deaths as of September 2022. Since the beginning of the pandemic, several variants of SARS-CoV-2 have emerged, with different infectivity and virulence. Several studies suggest an important role of neutrophils in SARS-Cov-2 infection severity, but data about direct activation of neutrophils by the virus is scarce. Here, we studied the in vitro activation of human neutrophils by SARS-CoV-2 variants of concern (VOCs). In our work, we show that upon stimulation with SARS-Cov-2 infectious particles, human healthy resting neutrophils upregulate activation markers, degranulate IL-8, produce Reactive Oxygen Species and release Neutrophil Extracellular Traps. Neutrophil activation was dependent on TLR7/8 and IRF3/STING. We then compared the activation potential of neutrophils by SARS-CoV-2 variants and showed a significantly increased activation by the Delta variant and a decreased activation by the Omicron variant as compared to the initial strain. In this study, we demonstrate that the SARS-Cov-2 virus can directly activate neutrophils in COVID-19 and that the different VOCs had differences in neutrophil activation intensity that mirror the differences of clinical severity. These data highlight the need to address neutrophil-virus interactions as a potential target for therapeutic intervention in SARS-CoV-2 infection.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , Neutrophils
10.
Sci Transl Med ; 14(671): eabo5795, 2022 Nov 16.
Article in English | MEDLINE | ID: covidwho-2119264

ABSTRACT

Interstitial lung disease and associated fibrosis occur in a proportion of individuals who have recovered from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection through unknown mechanisms. We studied individuals with severe coronavirus disease 2019 (COVID-19) after recovery from acute illness. Individuals with evidence of interstitial lung changes at 3 to 6 months after recovery had an up-regulated neutrophil-associated immune signature including increased chemokines, proteases, and markers of neutrophil extracellular traps that were detectable in the blood. Similar pathways were enriched in the upper airway with a concomitant increase in antiviral type I interferon signaling. Interaction analysis of the peripheral phosphoproteome identified enriched kinases critical for neutrophil inflammatory pathways. Evaluation of these individuals at 12 months after recovery indicated that a subset of the individuals had not yet achieved full normalization of radiological and functional changes. These data provide insight into mechanisms driving development of pulmonary sequelae during and after COVID-19 and provide a rational basis for development of targeted approaches to prevent long-term complications.


Subject(s)
COVID-19 , Extracellular Traps , Humans , SARS-CoV-2 , Neutrophils , Lung
11.
Cell Rep Med ; 3(10): 100779, 2022 10 18.
Article in English | MEDLINE | ID: covidwho-2096146

ABSTRACT

Mechanisms of neutrophil involvement in severe coronavirus disease 2019 (COVID-19) remain incompletely understood. Here, we collect longitudinal blood samples from 306 hospitalized COVID-19+ patients and 86 controls and perform bulk RNA sequencing of enriched neutrophils, plasma proteomics, and high-throughput antibody profiling to investigate relationships between neutrophil states and disease severity. We identify dynamic switches between six distinct neutrophil subtypes. At days 3 and 7 post-hospitalization, patients with severe disease display a granulocytic myeloid-derived suppressor cell-like gene expression signature, while patients with resolving disease show a neutrophil progenitor-like signature. Humoral responses are identified as potential drivers of neutrophil effector functions, with elevated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific immunoglobulin G1 (IgG1)-to-IgA1 ratios in plasma of severe patients who survived. In vitro experiments confirm that while patient-derived IgG antibodies induce phagocytosis in healthy donor neutrophils, IgA antibodies predominantly induce neutrophil cell death. Overall, our study demonstrates a dysregulated myelopoietic response in severe COVID-19 and a potential role for IgA-dominant responses contributing to mortality.


Subject(s)
COVID-19 , Humans , SARS-CoV-2 , Neutrophils , Immunoglobulin A , Immunoglobulin G , Phenotype
12.
PLoS One ; 17(10): e0276460, 2022.
Article in English | MEDLINE | ID: covidwho-2089429

ABSTRACT

Excessive neutrophil infiltration and dysfunction contribute to the progression and severity of hyper-inflammatory syndrome, such as in severe COVID19. In the current study, we re-analysed published scRNA-seq datasets of mouse and human neutrophils to classify and compare the transcriptional regulatory networks underlying neutrophil differentiation and inflammatory responses. Distinct sets of TF modules regulate neutrophil maturation, function, and inflammatory responses under the steady state and inflammatory conditions. In COVID19 patients, neutrophil activation was associated with the selective activation of inflammation-specific TF modules. SARS-CoV-2 RNA-positive neutrophils showed a higher expression of type I interferon response TF IRF7. Furthermore, IRF7 expression was abundant in neutrophils from severe patients in progression stage. Neutrophil-mediated inflammatory responses positively correlate with the expressional level of IRF7. Based on these results, we suggest that differential activation of activation-related TFs, such as IRF7 mediate neutrophil inflammatory responses during inflammation.


Subject(s)
COVID-19 , Neutrophils , Humans , COVID-19/genetics , COVID-19/metabolism , Inflammation/genetics , Interferon Type I/metabolism , Neutrophil Activation/genetics , Neutrophil Activation/physiology , Neutrophils/metabolism , RNA, Viral , RNA-Seq , SARS-CoV-2 , Single-Cell Analysis
13.
Clin Lab ; 68(10)2022 Oct 01.
Article in English | MEDLINE | ID: covidwho-2080864

ABSTRACT

BACKGROUND: Inflammatory processes activated by rapid viral replication of SARS-CoV-2 can play a key role in the pathogenesis of multiple organ damage and be responsible for the COVID-19 patients' dramatic outcomes and common abnormal laboratory findings. The aim of this study was to assess the correlation between various laboratory biomarkers, ferritin/transferrin ratio (FTR) and receiver operating characteristic (ROC) analysis in monitoring COVID-19 patients. METHODS: This observational study was conducted in three groups: healthy participants, non COVID-19 patients with COVID-19-like clinical signs, and COVID-19 patients (severe and non-severe). Biochemical (CRP, ferritin, transferrin and albumin) and hematological (WBC, lymphocytes) parameters were assessed by automated methods. Moreover, FTR and NLR markers were calculated in the three groups mentioned. Statistical analyses were done using R (version 4.1.0). ROC curve was used to validate the predictive value of parameters. RESULTS: The COVID-19 positive group had significantly higher NEU, CRP, ferritin, FTR values, while it's WBC, absolute counts of lymphocytes and albumin were significantly lower compared to the non-COVID-19 patients (p < 0.001). Serum ferritin and FTR level of the severe group was significantly higher than that of the non-severe group (p = 0.006 and (p = 0.011, respectively). The strongest correlation in all subjects showed between lymphocytopenia and increased NEU (r = -0.99, p < 0.001). The AUC values of WBC (0.95), lymphocytes (0.89), NEU (0.88), and NLR (0.88) were higher than CRP (0.64) or Ferritin (0.81). CONCLUSIONS: We recommend using FTR, WBC, and NLR changes as simple, useful, and inexpensive indicators in early detection of COVID-19 patients.


Subject(s)
COVID-19 , Albumins , Biomarkers , COVID-19/diagnosis , Ferritins , Humans , Neutrophils , Prognosis , ROC Curve , Retrospective Studies , SARS-CoV-2 , Transferrin
14.
J Leukoc Biol ; 111(6): 1159-1173, 2022 06.
Article in English | MEDLINE | ID: covidwho-2075036

ABSTRACT

Neutrophils play significant roles in immune homeostasis and as neutralizers of microbial infections. Recent evidence further suggests heterogeneity of neutrophil developmental and activation states that exert specialized effector functions during inflammatory disease conditions. Neutrophils can play multiple roles during viral infections, secreting inflammatory mediators and cytokines that contribute significantly to host defense and pathogenicity. However, their roles in viral immunity are not well understood. In this review, we present an overview of neutrophil heterogeneity and its impact on the course and severity of viral respiratory infectious diseases. We focus on the evidence demonstrating the crucial roles neutrophils play in the immune response toward respiratory infections, using influenza as a model. We further extend the understanding of neutrophil function with the studies pertaining to COVID-19 disease and its neutrophil-associated pathologies. Finally, we discuss the relevance of these results for future therapeutic options through targeting and regulating neutrophil-specific responses.


Subject(s)
COVID-19 , Virus Diseases , Cytokines , Humans , Inflammation Mediators , Neutrophil Activation , Neutrophils , Virus Diseases/pathology
15.
Int J Mol Sci ; 23(18)2022 Sep 09.
Article in English | MEDLINE | ID: covidwho-2071501

ABSTRACT

In SARS-CoV-2-infected humans, disease progression is often associated with acute respiratory distress syndrome involving severe lung injury, coagulopathy, and thrombosis of the alveolar capillaries. The pathogenesis of these pulmonary complications in COVID-19 patients has not been elucidated. Autopsy study of these patients showed SARS-CoV-2 virions in pulmonary vessels and sequestrated leukocytes infiltrates associated with endotheliopathy and microvascular thrombosis. Since SARS-CoV-2 enters and infects target cells by binding its spike (S) protein to cellular angiotensin-converting enzyme 2 (ACE2), and there is evidence that vascular endothelial cells and neutrophils express ACE2, we investigated the effect of S-proteins and cell-cell communication on primary human lung microvascular endothelial cells (HLMEC) and neutrophils expression of thrombogenic factors and the potential mechanisms. Using S-proteins of two different SARS-CoV-2 variants (Wuhan and Delta), we demonstrate that exposure of HLMEC or neutrophils to S-proteins, co-culture of HLMEC exposed to S-proteins with non-exposed neutrophils, or co-culture of neutrophils exposed to S-proteins with non-exposed HLMEC induced transcriptional upregulation of tissue factor (TF), significantly increased the expression and secretion of factor (F)-V, thrombin, and fibrinogen and inhibited tissue factor pathway inhibitor (TFPI), the primary regulator of the extrinsic pathway of blood coagulation, in both cell types. Recombinant (r)TFPI and a thiol blocker (5,5'-dithio-bis-(2-nitrobenzoic acid)) prevented S-protein-induced expression and secretion of Factor-V, thrombin, and fibrinogen. Thrombomodulin blocked S-protein-induced expression and secretion of fibrinogen but had no effect on S-protein-induced expression of Factor-V or thrombin. These results suggests that following SARS-CoV-2 contact with the pulmonary endothelium or neutrophils and endothelial-neutrophil interactions, viral S-proteins induce coagulopathy via the TF pathway and mechanisms involving functional thiol groups. These findings suggest that using rTFPI and/or thiol-based drugs could be a viable therapeutic strategy against SARS-CoV-2-induced coagulopathy and thrombosis.


Subject(s)
Blood Coagulation Disorders , COVID-19 , Thrombosis , Angiotensin-Converting Enzyme 2 , Cell Communication , Endothelial Cells/metabolism , Endothelium/metabolism , Fibrinogen , Humans , Lipoproteins , Lung/metabolism , Neutrophils/metabolism , SARS-CoV-2 , Spike Glycoprotein, Coronavirus/metabolism , Sulfhydryl Compounds , Thrombin , Thrombomodulin , Thromboplastin , Thrombosis/etiology
16.
Front Immunol ; 13: 918476, 2022.
Article in English | MEDLINE | ID: covidwho-2071085

ABSTRACT

Background: Deep venous thrombosis (DVT) highly occurs in patients with severe COVID-19 and probably accounted for their high mortality. DVT formation is a time-dependent inflammatory process in which NETosis plays an important role. However, whether ginsenoside Rg5 from species of Panax genus could alleviate DVT and its underlying mechanism has not been elucidated. Methods: The interaction between Rg5 and P2RY12 was studied by molecular docking, molecular dynamics, surface plasmon resonance (SPR), and molecular biology assays. The preventive effect of Rg5 on DVT was evaluated in inferior vena cava stasis-induced mice, and immunocytochemistry, Western blot, and calcium flux assay were performed in neutrophils from bone marrow to explore the mechanism of Rg5 in NETosis via P2RY12. Results: Rg5 allosterically interacted with P2RY12, formed stable complex, and antagonized its activity via residue E188 and R265. Rg5 ameliorated the formation of thrombus in DVT mice; accompanied by decreased release of Interleukin (IL)-6, IL-1ß, and tumor necrosis factor-α in plasma; and suppressed neutrophil infiltration and neutrophil extracellular trap (NET) release. In lipopolysaccharide- and platelet-activating factor-induced neutrophils, Rg5 reduced inflammatory responses via inhibiting the activation of ERK/NF-κB signaling pathway while decreasing cellular Ca2+ concentration, thus reducing the activity and expression of peptidyl arginine deiminase 4 to prevent NETosis. The inhibitory effect on neutrophil activity was dependent on P2RY12. Conclusions: Rg5 could attenuate experimental DVT by counteracting NETosis and inflammatory response in neutrophils via P2RY12, which may pave the road for its clinical application in the prevention of DVT-related disorders.


Subject(s)
COVID-19 , Venous Thrombosis , Animals , Ginsenosides , Mice , Molecular Docking Simulation , Neutrophils
17.
PLoS One ; 17(10): e0275391, 2022.
Article in English | MEDLINE | ID: covidwho-2054380

ABSTRACT

INTRODUCTION: In hospitalized COVID-19, neutrophil-to-lymphocyte ratio (NLR) and serum creatinine is sometimes measured under assumption they predict disease severity and mortality. We determined the potential value of NLR and serum creatinine as predictors of disease severity and mortality in COVID-19. METHODS: Prospective cohort study of COVID-19 patients admitted to premier COVID-19 treatment hospitals in Ethiopia. Predictive capability of biomarkers in progression and prognosis of COVID-19 was analyzed using receiver operating characteristics. Survival of COVID-19 patients with different biomarker levels was computed. Logistic regression assessed associations between disease severity and mortality on NLR and serum creatinine adjusted for odds ratio (AOR). RESULTS: The study enrolled 126 adults with severe (n = 68) or mild/moderate (n = 58) COVID-19, with median age 50 [interquartile range (IQR 20-86)]; 57.1% males. The NLR value was significantly higher in severe cases [6.68 (IQR 3.03-12.21)] compared to the mild/moderate [3.23 (IQR 2.09-5.39)], with the NLR value markedly associated with disease severity (p<0.001). Mortality was higher in severe cases [13 (19.1%)] compared to mild/moderate cases [2 (3.4%)] (p = 0.007). The NLR value was significantly higher in non-survivors [15.17 (IQR 5.13-22.5)] compared to survivors [4.26 (IQR 2.40-7.90)] (p = 0.002). Serum creatinine was significantly elevated in severe cases [34 (50%)] compared with mild/moderate [11 (19%)] (p<0.001). Disease severity [AOR 6.58, 95%CI (1.29-33.56), p = 0.023] and NLR [AOR 1.07, 95%CI (1.02-1.12), p = 0.004)] might be associated with death. NLR had a sensitivity and specificity of 69.1% and 60.3% as predictor of disease severity (cut-off >4.08), and 86.7% and 55.9% as prognostic marker of mortality (cut-off >4.63). CONCLUSION: In COVID-19, NLR is a biomarker with only modest accuracy for predicting disease severity and mortality. Still, patients with NLR >4.63 are more likely to die. Monitoring of this biomarker at the earliest stage of the disease may predict outcome. Additionally, high creatinine seems related to disease severity and mortality.


Subject(s)
COVID-19 , Neutrophils , Adult , Biomarkers , COVID-19/drug therapy , Creatinine , Female , Humans , Lymphocytes , Male , Middle Aged , Point-of-Care Systems , Prognosis , Prospective Studies , Retrospective Studies , Severity of Illness Index
18.
Cells ; 11(18)2022 09 16.
Article in English | MEDLINE | ID: covidwho-2043594

ABSTRACT

Rationale: Infection with the SARS-CoV2 virus is associated with elevated neutrophil counts. Evidence of neutrophil dysfunction in COVID-19 is based on transcriptomics or single functional assays. Cell functions are interwoven pathways, and understanding the effect across the spectrum of neutrophil function may identify therapeutic targets. Objectives: Examine neutrophil phenotype and function in 41 hospitalised, non-ICU COVID-19 patients versus 23 age-matched controls (AMC) and 26 community acquired pneumonia patients (CAP). Methods: Isolated neutrophils underwent ex vivo analyses for migration, bacterial phagocytosis, ROS generation, NETosis and receptor expression. Circulating DNAse 1 activity, levels of cfDNA, MPO, VEGF, IL-6 and sTNFRI were measured and correlated to clinical outcome. Serial sampling on day three to five post hospitalization were also measured. The effect of ex vivo PI3K inhibition was measured in a further cohort of 18 COVID-19 patients. Results: Compared to AMC and CAP, COVID-19 neutrophils demonstrated elevated transmigration (p = 0.0397) and NETosis (p = 0.0332), and impaired phagocytosis (p = 0.0036) associated with impaired ROS generation (p < 0.0001). The percentage of CD54+ neutrophils (p < 0.001) was significantly increased, while surface expression of CD11b (p = 0.0014) and PD-L1 (p = 0.006) were significantly decreased in COVID-19. COVID-19 and CAP patients showed increased systemic markers of NETosis including increased cfDNA (p = 0.0396) and impaired DNAse activity (p < 0.0001). The ex vivo inhibition of PI3K γ and δ reduced NET release by COVID-19 neutrophils (p = 0.0129). Conclusions: COVID-19 is associated with neutrophil dysfunction across all main effector functions, with altered phenotype, elevated migration and NETosis, and impaired antimicrobial responses. These changes highlight that targeting neutrophil function may help modulate COVID-19 severity.


Subject(s)
COVID-19 , Neutrophils , B7-H1 Antigen , COVID-19/immunology , Cell-Free Nucleic Acids , Deoxyribonucleases , Humans , Interleukin-6/pharmacology , Neutrophils/cytology , Phenotype , Phosphatidylinositol 3-Kinases , Reactive Oxygen Species/metabolism , SARS-CoV-2
19.
Front Immunol ; 13: 996637, 2022.
Article in English | MEDLINE | ID: covidwho-2043454

ABSTRACT

Increased neutrophils and elevated level of circulating calprotectin are hallmarks of severe COVID-19 and they contribute to the dysregulated immune responses and cytokine storm in susceptible patients. However, the precise mechanism controlling calprotectin production during SARS-CoV-2 infection remains elusive. In this study, we showed that Dok3 adaptor restrains calprotectin production by neutrophils in response to SARS-CoV-2 spike (S) protein engagement of TLR4. Dok3 recruits SHP-2 to mediate the de-phosphorylation of MyD88 at Y257, thereby attenuating downstream JAK2-STAT3 signaling and calprotectin production. Blocking of TLR4, JAK2 and STAT3 signaling could prevent excessive production of calprotectin by Dok3-/- neutrophils, revealing new targets for potential COVID-19 therapy. As S protein from SARS-CoV-2 Delta and Omicron variants can activate TLR4-driven calprotectin production in Dok3-/- neutrophils, our study suggests that targeting calprotectin production may be an effective strategy to combat severe COVID-19 manifestations associated with these emerging variants.


Subject(s)
Adaptor Proteins, Signal Transducing , COVID-19 , Spike Glycoprotein, Coronavirus , Adaptor Proteins, Signal Transducing/metabolism , Humans , Leukocyte L1 Antigen Complex , Myeloid Differentiation Factor 88/metabolism , Neutrophils/metabolism , SARS-CoV-2 , Toll-Like Receptor 4/metabolism
20.
Front Immunol ; 13: 995886, 2022.
Article in English | MEDLINE | ID: covidwho-2043453

ABSTRACT

Kawasaki disease (KD), a multisystem inflammatory syndrome that occurs in children, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2 or COVID-19) may share some overlapping mechanisms. The purpose of this study was to analyze the differences in single-cell RNA sequencing between KD and COVID-19. We performed single-cell RNA sequencing in KD patients (within 24 hours before IVIG treatment) and age-matched fever controls. The single-cell RNA sequencing data of COVID-19, influenza, and health controls were downloaded from the Sequence Read Archive (GSE149689/PRJNA629752). In total, 22 single-cell RNA sequencing data with 102,355 nuclei were enrolled in this study. After performing hierarchical and functional clustering analyses, two enriched gene clusters demonstrated similar patterns in severe COVID-19 and KD, heightened neutrophil activation, and decreased MHC class II expression. Furthermore, comparable dysregulation of neutrophilic granulopoiesis representing two pronounced hyperinflammatory states was demonstrated, which play a critical role in the overactivated and defective aging program of granulocytes, in patients with KD as well as those with severe COVID-19. In conclusion, both neutrophil activation and MHC class II reduction play a crucial role and thus may provide potential treatment targets for KD and severe COVID-19.


Subject(s)
COVID-19 , Mucocutaneous Lymph Node Syndrome , COVID-19/complications , Child , Humans , Immunoglobulins, Intravenous , Neutrophils , SARS-CoV-2 , Systemic Inflammatory Response Syndrome
SELECTION OF CITATIONS
SEARCH DETAIL