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1.
Front Immunol ; 13: 842150, 2022.
Article in English | MEDLINE | ID: covidwho-1779941

ABSTRACT

Although it is now widely accepted that host inflammatory response contributes to COVID-19 immunopathogenesis, the pathways and mechanisms driving disease severity and clinical outcome remain poorly understood. In the effort to identify key soluble mediators that characterize life-threatening COVID-19, we quantified 62 cytokines, chemokines and other factors involved in inflammation and immunity in plasma samples, collected at hospital admission, from 80 hospitalized patients with severe COVID-19 disease who were stratified on the basis of clinical outcome (mechanical ventilation or death by day 28). Our data confirm that age, as well as neutrophilia, lymphocytopenia, procalcitonin, D-dimer and lactate dehydrogenase are strongly associated with the risk of fatal COVID-19. In addition, we found that cytokines related to TH2 regulations (IL-4, IL-13, IL-33), cell metabolism (lep, lep-R) and interferons (IFNα, IFNß, IFNγ) were also predictive of life-threatening COVID-19.


Subject(s)
COVID-19 , Cytokines , Chemokines , Humans , Interferons , SARS-CoV-2
2.
EuropePMC;
Preprint in English | EuropePMC | ID: ppcovidwho-328786

ABSTRACT

Aging is a major risk factor for developing severe COVID-19, but few detailed data are available concerning immunological changes after infection in aged individuals. Here we describe main immune characteristics in 31 patients with severe SARS-CoV-2 infection who were >70 years old, compared to 33 subjects <60 years of age. Differences in plasma levels of 62 cytokines, landscape of peripheral blood mononuclear cells, T cell repertoire, transcriptome of central memory CD4 + T cells, specific antibodies are reported along with features of lung macrophages. Elderly subjects have higher levels of pro-inflammatory cytokines, more circulating plasmablasts, reduced plasmatic level of anti-S and anti-RBD IgG3 antibodies, lower proportions of central memory CD4 + T cells, more immature monocytes and CD56 + pro-inflammatory monocytes, lower percentages of circulating follicular helper T cells (cTfh), antigen-specific cTfh cells with a less activated transcriptomic profile, lung resident activated macrophages that promote collagen deposition and fibrosis. Our study underlines the importance of inflammation in the response to SARS-CoV-2 and suggests that inflammaging, coupled with the inability to mount a proper anti-viral response, could exacerbate disease severity and the worst clinical outcome in old patients.

3.
EuropePMC; 2021.
Preprint in English | EuropePMC | ID: ppcovidwho-306885

ABSTRACT

In 14 pregnant women who had asymptomatic or paucisymptomatic SARS-CoV-2 infection, we performed a detailed 38-parameter analysis of peripheral blood mononuclear cells by mass cytometry, studied the expression of T-cell master regular genes, investigated cell proliferation and cytokine production, and measured plasma levels of 62 cytokines. No patient showed lymphopenia or gross alterations of white blood cells. Unsupervised analyses revealed that most immune parameters were similar in patients and uninfected controls, apart from an increase in low density neutrophils in SARS-CoV-2 positive women. Also, patients did not show altered plasma levels of interleukin-6 or other main inflammatory molecules, but displayed significant increases of anti-inflammatory cytokines such as IL-1RA, IL-10 and IL-19, and decreased levels of IL-17, PD-L1 and D-dimer. The endogenous control of inflammation, as evidenced by plasma levels of soluble molecules, could be a strategy used during pregnancy to avoid virus-induced damages and maintain a normal immune response.

4.
EuropePMC; 2020.
Preprint in English | EuropePMC | ID: ppcovidwho-306884

ABSTRACT

We have deeply investigated T cell compartment, plasma cytokines and cells producing cytokines in patients affected by Covid-19. At admission, patients were lymphopenic;in all of them SARS-CoV-2 was detected in a nasopharyngeal swab specimen by real-time RT-PCR, and pneumonia was subsequently confirmed by X-rays.Detailed 18-parameter flow cytometry was performed in 21 patients and 13 controls. Coupling polychromatic cytometry with unsupervised data analysis, we found that patients show an increased amount of CD4+ T lymphocytes that were activated, exhausted, stem memory or Treg. Similar results concerning activation and exhaustion were found in the CD8+ T cell compartment, within which the differences were even greater.Measuring plasma level of 31 cytokines linked to inflammation revealed that Covid-19 showed a dramatic increase of several molecules, such as TH1 and TH2 cytokines, chemokines, galectins, pro- and anti-inflammatory mediators, confirming the importance of a massive immune activation causing the cytokine storm. Then, intracellular staining detecting the simultaneous production of different cytokines after a para-physiologic stimulus given by anti-CD3/CD28 mAbs revealed not only a high capacity to produce a variety of molecules, including TNF-a, IFN-g and IL-2, but also a significant skewing of CD4+ T cells towards the TH17 phenotype.A therapeutic approach now exists based on the administration of drugs that block IL-6 pathway, and is now consistently improving the course of the disease. IL-17 is crucial in recruiting and activating neutrophils, cells that can migrate to the lung and are heavily involved in the pathogenesis of Covid-19. We show here that a significant skewing of activated T cells towards TH17 functional phenotype exists in Covid-19 patients. Thus, we suggest that blocking IL-17 pathway by already available biological drugs that are used to treat different pathologies could be a novel, additional strategy to improve the health of patients infected by SARS-CoV-2.

5.
Eur J Immunol ; 52(3): 484-502, 2022 03.
Article in English | MEDLINE | ID: covidwho-1555185

ABSTRACT

To better understand the mechanisms at the basis of neutrophil functions during SARS-CoV-2, we studied patients with severe COVID-19 pneumonia. They had high blood proportion of degranulated neutrophils and elevated plasma levels of myeloperoxidase (MPO), elastase, and MPO-DNA complexes, which are typical markers of neutrophil extracellular traps (NET). Their neutrophils display dysfunctional mitochondria, defective oxidative burst, increased glycolysis, glycogen accumulation in the cytoplasm, and increase glycogenolysis. Hypoxia-inducible factor 1α (ΗΙF-1α) is stabilized in such cells, and it controls the level of glycogen phosphorylase L (PYGL), a key enzyme in glycogenolysis. Inhibiting PYGL abolishes the ability of neutrophils to produce NET. Patients displayed significant increases of plasma levels of molecules involved in the regulation of neutrophils' function including CCL2, CXCL10, CCL20, IL-18, IL-3, IL-6, G-CSF, GM-CSF, IFN-γ. Our data suggest that metabolic remodelling is vital for the formation of NET and for boosting neutrophil inflammatory response, thus, suggesting that modulating ΗΙF-1α or PYGL could represent a novel approach for innovative therapies.


Subject(s)
COVID-19/immunology , COVID-19/metabolism , Neutrophils/immunology , Neutrophils/metabolism , SARS-CoV-2 , Adult , Aged , Aged, 80 and over , COVID-19/blood , Case-Control Studies , Cohort Studies , Cytokines/blood , Extracellular Traps/immunology , Extracellular Traps/metabolism , Female , Glycogen Phosphorylase, Liver Form/blood , Granulocytes/immunology , Granulocytes/metabolism , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/blood , Male , Metabolic Networks and Pathways/genetics , Metabolic Networks and Pathways/immunology , Middle Aged , Neutrophil Activation , Peroxidase/blood , Respiratory Burst , Severity of Illness Index
8.
Nat Commun ; 12(1): 4677, 2021 07 29.
Article in English | MEDLINE | ID: covidwho-1387356

ABSTRACT

SARS-CoV-2 infection can affect all human beings, including pregnant women. Thus, understanding the immunological changes induced by the virus during pregnancy is nowadays of pivotal importance. Here, using peripheral blood from 14 pregnant women with asymptomatic or mild SARS-CoV-2 infection, we investigate cell proliferation and cytokine production, measure plasma levels of 62 cytokines, and perform a 38-parameter mass cytometry analysis. Our results show an increase in low density neutrophils but no lymphopenia or gross alterations of white blood cells, which display normal levels of differentiation, activation or exhaustion markers and show well preserved functionality. Meanwhile, the plasma levels of anti-inflammatory cytokines such as interleukin (IL)-1RA, IL-10 and IL-19 are increased, those of IL-17, PD-L1 and D-dimer are decreased, but IL-6 and other inflammatory molecules remain unchanged. Our profiling of antiviral immune responses may thus help develop therapeutic strategies to avoid virus-induced damages during pregnancy.


Subject(s)
COVID-19/immunology , Cytokines/blood , Inflammation/immunology , Pregnancy Complications, Infectious/immunology , Pregnancy Complications, Infectious/virology , SARS-CoV-2/immunology , Adolescent , Adult , Asymptomatic Infections , Biomarkers/blood , COVID-19/blood , COVID-19/virology , Case-Control Studies , Cross-Sectional Studies , Cytokines/immunology , Female , Humans , Inflammation/blood , Inflammation/prevention & control , Inflammation/virology , Middle Aged , Pregnancy , Pregnancy Complications, Infectious/blood , SARS-CoV-2/isolation & purification , Young Adult
9.
Cells ; 10(7)2021 06 23.
Article in English | MEDLINE | ID: covidwho-1285369

ABSTRACT

Cell death mechanisms are crucial to maintain an appropriate environment for the functionality of healthy cells. However, during viral infections, dysregulation of these processes can be present and can participate in the pathogenetic mechanisms of the disease. In this review, we describe some features of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and some immunopathogenic mechanisms characterizing the present coronavirus disease (COVID-19). Lymphopenia and monocytopenia are important contributors to COVID-19 immunopathogenesis. The fine mechanisms underlying these phenomena are still unknown, and several hypotheses have been raised, some of which assign a role to cell death as far as the reduction of specific types of immune cells is concerned. Thus, we discuss three major pathways such as apoptosis, necroptosis, and pyroptosis, and suggest that all of them likely occur simultaneously in COVID-19 patients. We describe that SARS-CoV-2 can have both a direct and an indirect role in inducing cell death. Indeed, on the one hand, cell death can be caused by the virus entry into cells, on the other, the excessive concentration of cytokines and chemokines, a process that is known as a COVID-19-related cytokine storm, exerts deleterious effects on circulating immune cells. However, the overall knowledge of these mechanisms is still scarce and further studies are needed to delineate new therapeutic strategies.


Subject(s)
COVID-19/pathology , Cell Death/physiology , SARS-CoV-2/pathogenicity , Apoptosis/physiology , COVID-19/immunology , COVID-19/virology , Cytokine Release Syndrome/immunology , Cytokine Release Syndrome/pathology , Cytokines/metabolism , Humans , Necroptosis/physiology , Virus Internalization
10.
EMBO Mol Med ; 12(12): e13001, 2020 12 07.
Article in English | MEDLINE | ID: covidwho-881540

ABSTRACT

In patients infected by SARS-CoV-2 who experience an exaggerated inflammation leading to pneumonia, monocytes likely play a major role but have received poor attention. Thus, we analyzed peripheral blood monocytes from patients with COVID-19 pneumonia and found that these cells show signs of altered bioenergetics and mitochondrial dysfunction, had a reduced basal and maximal respiration, reduced spare respiratory capacity, and decreased proton leak. Basal extracellular acidification rate was also diminished, suggesting reduced capability to perform aerobic glycolysis. Although COVID-19 monocytes had a reduced ability to perform oxidative burst, they were still capable of producing TNF and IFN-γ in vitro. A significantly high amount of monocytes had depolarized mitochondria and abnormal mitochondrial ultrastructure. A redistribution of monocyte subsets, with a significant expansion of intermediate/pro-inflammatory cells, and high amounts of immature monocytes were found, along with a concomitant compression of classical monocytes, and an increased expression of inhibitory checkpoints like PD-1/PD-L1. High plasma levels of several inflammatory cytokines and chemokines, including GM-CSF, IL-18, CCL2, CXCL10, and osteopontin, finally confirm the importance of monocytes in COVID-19 immunopathogenesis.


Subject(s)
COVID-19/pathology , Energy Metabolism/physiology , Mitochondria/metabolism , Monocytes/metabolism , Adult , Aged , Aged, 80 and over , COVID-19/virology , Case-Control Studies , Chemokines/blood , Cytokines/blood , Female , Humans , Male , Middle Aged , Mitochondria/ultrastructure , Monocytes/cytology , Programmed Cell Death 1 Receptor/genetics , Programmed Cell Death 1 Receptor/metabolism , SARS-CoV-2/isolation & purification
11.
Eur J Immunol ; 50(9): 1283-1294, 2020 09.
Article in English | MEDLINE | ID: covidwho-670172

ABSTRACT

Studies on the interactions between SARS-CoV-2 and humoral immunity are fundamental to elaborate effective therapies including vaccines. We used polychromatic flow cytometry, coupled with unsupervised data analysis and principal component analysis (PCA), to interrogate B cells in untreated patients with COVID-19 pneumonia. COVID-19 patients displayed normal plasma levels of the main immunoglobulin classes, of antibodies against common antigens or against antigens present in common vaccines. However, we found a decreased number of total and naïve B cells, along with decreased percentages and numbers of memory switched and unswitched B cells. On the contrary, IgM+ and IgM- plasmablasts were significantly increased. In vitro cell activation revealed that B lymphocytes showed a normal proliferation index and number of dividing cells per cycle. PCA indicated that B-cell number, naive and memory B cells but not plasmablasts clustered with patients who were discharged, while plasma IgM level, C-reactive protein, D-dimer, and SOFA score with those who died. In patients with pneumonia, the derangement of the B-cell compartment could be one of the causes of the immunological failure to control SARS-Cov2, have a relevant influence on several pathways, organs and systems, and must be considered to develop vaccine strategies.


Subject(s)
Antibodies, Viral/blood , B-Lymphocytes/immunology , Betacoronavirus/pathogenicity , Coronavirus Infections/immunology , Immunoglobulin Isotypes/blood , Lung/immunology , Pneumonia, Viral/immunology , Adult , Aged , Aged, 80 and over , Antibodies, Viral/classification , B-Lymphocytes/virology , Betacoronavirus/immunology , C-Reactive Protein/immunology , COVID-19 , Case-Control Studies , Cell Proliferation , Coronavirus Infections/mortality , Coronavirus Infections/pathology , Coronavirus Infections/virology , Cross-Sectional Studies , Cytokines/genetics , Cytokines/immunology , Female , Fibrin Fibrinogen Degradation Products/immunology , Humans , Immunity, Humoral , Immunologic Memory , Lung/pathology , Lung/virology , Lymphocyte Activation , Lymphocyte Count , Male , Middle Aged , Organ Dysfunction Scores , Pandemics , Pneumonia, Viral/mortality , Pneumonia, Viral/pathology , Pneumonia, Viral/virology , Primary Cell Culture , SARS-CoV-2 , Severity of Illness Index , Survival Analysis
12.
Nat Commun ; 11(1): 3434, 2020 07 06.
Article in English | MEDLINE | ID: covidwho-631255

ABSTRACT

The immune system of patients infected by SARS-CoV-2 is severely impaired. Detailed investigation of T cells and cytokine production in patients affected by COVID-19 pneumonia are urgently required. Here we show that, compared with healthy controls, COVID-19 patients' T cell compartment displays several alterations involving naïve, central memory, effector memory and terminally differentiated cells, as well as regulatory T cells and PD1+CD57+ exhausted T cells. Significant alterations exist also in several lineage-specifying transcription factors and chemokine receptors. Terminally differentiated T cells from patients proliferate less than those from healthy controls, whereas their mitochondria functionality is similar in CD4+ T cells from both groups. Patients display significant increases of proinflammatory or anti-inflammatory cytokines, including T helper type-1 and type-2 cytokines, chemokines and galectins; their lymphocytes produce more tumor necrosis factor (TNF), interferon-γ, interleukin (IL)-2 and IL-17, with the last observation implying that blocking IL-17 could provide a novel therapeutic strategy for COVID-19.


Subject(s)
Betacoronavirus/immunology , CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , Coronavirus Infections/immunology , Pneumonia, Viral/immunology , T-Lymphocyte Subsets/immunology , Adult , Aged , Aged, 80 and over , Biomarkers/metabolism , CD4-Positive T-Lymphocytes/metabolism , CD4-Positive T-Lymphocytes/pathology , CD8-Positive T-Lymphocytes/metabolism , CD8-Positive T-Lymphocytes/pathology , COVID-19 , Cellular Senescence , Coronavirus Infections/blood , Coronavirus Infections/pathology , Cytokine Release Syndrome , Cytokines/immunology , Cytokines/metabolism , Female , Humans , Immunologic Memory , Italy/epidemiology , Lymphocyte Activation , Lymphocyte Count , Male , Middle Aged , Pandemics , Pneumonia, Viral/blood , Pneumonia, Viral/pathology , SARS-CoV-2 , T-Lymphocyte Subsets/metabolism , T-Lymphocyte Subsets/pathology , Th17 Cells/immunology , Th17 Cells/metabolism , Th17 Cells/pathology
13.
Cytometry A ; 97(7): 668-673, 2020 07.
Article in English | MEDLINE | ID: covidwho-47563

ABSTRACT

The pandemic caused by severe acute respiratory syndrome coronavirus 2 heavily involves all those working in a laboratory. Samples from known infected patients or donors who are considered healthy can arrive, and a colleague might be asymptomatic but able to transmit the virus. Working in a clinical laboratory is posing several safety challenges. Few years ago, International Society for Advancement of Cytometry published guidelines to safely analyze and sort human samples that were revised in these days. We describe the procedures that we have been following since the first patient appeared in Italy, which have only slightly modified our standard one, being all human samples associated with risks. © 2020 International Society for Advancement of Cytometry.


Subject(s)
Betacoronavirus , Coronavirus Infections/blood , Medical Laboratory Personnel , Occupational Health , Pneumonia, Viral/blood , Specimen Handling/methods , COVID-19 , Cytokine Release Syndrome/blood , Flow Cytometry/methods , Humans , Italy , Masks , Occupational Exposure/prevention & control , Pandemics , Protective Clothing , SARS-CoV-2
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