Integrated cytokine and metabolite analysis reveals immunometabolic reprogramming in COVID-19 patients with therapeutic implications

AbstractCytokine release syndrome (CRS) is a major cause of the multi-organ injury and fatal outcome induced by SARS-CoV-2 infection in severe COVID-19 patients. Metabolism can modulate the immune responses against infectious diseases, yet our understanding remains limited on how host metabolism correlates with inflammatory responses and affects cytokine release in COVID-19 patients. Here we perform both metabolomics and cytokine/chemokine profiling on serum samples from healthy controls, mild and severe COVID-19 patients, and delineate their global metabolic and immune response landscape. Correlation analyses show tight associations between metabolites and proinflammatory cytokines/chemokines, such as IL-6, M-CSF, IL-1α, IL-1β, and imply a potential regulatory crosstalk between arginine, tryptophan, purine metabolism and hyperinflammation. Importantly, we also demonstrate that targeting metabolism markedly modulates the proinflammatory cytokines release by peripheral blood mononuclear cells isolated from SARS-CoV-2-infected rhesus macaques ex vivo, hinting that exploiting metabolic alterations may be a potential strategy for treating fatal CRS in COVID-19.

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Host metabolite-cytokine correlation landscape in SARS-CoV-2 infection

10.21203/rs.3.rs-91866/v1 ◽

2020 ◽

Author(s):

Nan Xiao ◽

Meng Nie ◽

Huanhuan Pang ◽

Bohong Wang ◽

Jieli Hu ◽

...

Keyword(s):

Proinflammatory Cytokines ◽

Ex Vivo ◽

Rhesus Macaques ◽

Fatal Outcome ◽

Organ Injury ◽

Cytokine Release ◽

Serum Samples ◽

Cytokines And Chemokines ◽

Novel Strategy ◽

Host Metabolism

Abstract The systemic cytokine release syndrome (CRS) is a major cause of the multi-organ injury and fatal outcome induced by SARS-CoV-2 infection in severe COVID-19 patients. It has been well-known that metabolism plays a role in modulating the immune responses in infectious diseases. Yet, how the host metabolism correlates with CRS in COVID-19 patients and how the perturbed metabolites affect the cytokine release remains unclear. Here, we performed both metabolomics and cytokine/chemokine profiling on serum samples from the same cohort of healthy controls, mild and severe COVID-19 patients and delineated the global metabolic and immune response landscape along disease progression. Intriguingly, the correlation analysis revealed the tight link between metabolites and proinflammatory cytokines and chemokines, such as IL-6, M-CSF, IL-1α, IL-1β, implying the potential regulatory role of arginine metabolism, tryptophan metabolism, and purine metabolism in hyperinflammation. Importantly, we demonstrated that targeting metabolism markedly modulated the proinflammatory cytokines release by PBMCs isolated from SARS-CoV-2-infected rhesus macaques ex vivo. Beyond providing a comprehensive resource of metabolism and immunology data of SARS-CoV-2 infection, our study showed that metabolic alterations can be potentially exploited to develop novel strategy for the treatment of fatal CRS in COVID-19.

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Organ Injury and Cytokine Release Caused by Peptidoglycan Are Dependent on the Structural Integrity of the Glycan Chain

Infection and Immunity ◽

10.1128/iai.72.3.1311-1317.2004 ◽

2004 ◽

Vol 72 (3) ◽

pp. 1311-1317 ◽

Cited By ~ 19

Author(s):

Anders E. Myhre ◽

Jon Fredrik Stuestøl ◽

Maria K. Dahle ◽

Gunhild Øverland ◽

Christoph Thiemermann ◽

...

Keyword(s):

Structural Integrity ◽

Inflammatory Responses ◽

Ex Vivo ◽

Organ Injury ◽

Enzyme Linked Immunosorbent Assay ◽

Cytokine Release ◽

Ex Vivo Model ◽

Glycan Chain ◽

Tnf Α ◽

Glutamyl Transferase

ABSTRACT Several studies have implicated a role of peptidoglycan (PepG) as a pathogenicity factor in sepsis and organ injury, in part by initiating the release of inflammatory mediators. We wanted to elucidate the structural requirements of PepG to trigger inflammatory responses and organ injury. Injection of native PepG into anesthetized rats caused moderate but significant increases in the levels of alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transferase, and bilirubin (markers of hepatic injury and/or dysfunction) and creatinine and urea (markers of renal dysfunction) in serum, whereas PepG pretreated with muramidase to digest the glycan backbone failed to do this. In an ex vivo model of human blood, PepG containing different amino acids induced similar levels of the cytokines tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), IL-8, and IL-10, as determined by plasma analyses (enzyme-linked immunosorbent assay). Hydrolysis of the Staphylococcus aureus cross-bridge with lysostaphin resulted in moderately reduced release of TNF-α, IL-6, IL-8, and IL-10, whereas muramidase digestion nearly abolished the ability to induce cytokine release and IL-6 mRNA accumulation in CD14+ monocytes compared to intact PepG. However, additional experiments showed that muramidase-treated PepG synergized with lipopolysaccharide to induce TNF-α and IL-10 release in whole blood, despite its lack of inflammatory activity when administered alone. Based on these studies, we hypothesize that the structural integrity of the glycan chain of the PepG molecule is very important for the pathogenic effects of PepG. The amino acid composition of PepG, however, does not seem to be essential for the inflammatory properties of the molecule.

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An Updated Review of the Diagnostic Methods in Delayed Drug Hypersensitivity

Frontiers in Pharmacology ◽

10.3389/fphar.2020.573573 ◽

2021 ◽

Vol 11 ◽

Author(s):

Ana Copaescu ◽

Andrew Gibson ◽

Yueran Li ◽

Jason A. Trubiano ◽

Elizabeth J. Phillips

Keyword(s):

Mononuclear Cells ◽

Ex Vivo ◽

Drug Hypersensitivity ◽

Lymphocyte Transformation ◽

Diagnostic Methods ◽

Drug Discontinuation ◽

Hypersensitivity Reactions ◽

Cytokine Release ◽

Peripheral Blood Mononuclear ◽

Intradermal Testing

Delayed drug hypersensitivity reactions are clinically diverse reactions that vary from isolated benign skin conditions that remit quickly with no or symptomatic treatment, drug discontinuation or even continued drug treatment, to the other extreme of severe cutaneous adverse reactions (SCARs) that are associated with presumed life-long memory T-cell responses, significant acute and long-term morbidity and mortality. Diagnostic “in clinic” approaches to delayed hypersensitivity reactions have included patch testing (PT), delayed intradermal testing (IDT) and drug challenges for milder reactions. Patch and IDT are, in general, performed no sooner than 4–6 weeks after resolution of the acute reaction at the maximum non-irritating concentrations. Functional in vitro and ex vivo assays have largely remained the province of research laboratories and include lymphocyte transformation test (LTT) and cytokine release enzyme linked ImmunoSpot (ELISpot) assay, an emerging diagnostic tool which uses cytokine release, typically IFN-γ, after the patient’s peripheral blood mononuclear cells are stimulated with the suspected drug(s). Genetic markers such as human leukocyte antigen have shown recent promise for both pre-prescription screening as well as pre-emptive and diagnostic testing strategies.

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Boosted Pro-Inflammatory Activity in Human PBMCs by Lipopolysaccharide and SARS-CoV-2 Spike Protein Is Regulated by α-1 Antitrypsin

International Journal of Molecular Sciences ◽

10.3390/ijms22157941 ◽

2021 ◽

Vol 22 (15) ◽

pp. 7941

Author(s):

Srinu Tumpara ◽

Anna R. Gründing ◽

Kokilavani Sivaraman ◽

Sabine Wrenger ◽

Beata Olejnicka ◽

...

Keyword(s):

Human Plasma ◽

Mononuclear Cells ◽

Inflammatory Responses ◽

Ex Vivo ◽

Statistical Significance ◽

Spike Protein ◽

Human Pbmcs ◽

Experimental Conditions ◽

Peripheral Blood Mononuclear ◽

Anti Inflammatory

For the treatment of severe COVID-19, supplementation with human plasma-purified α-1 antitrypsin (AAT) to patients is currently considered. AAT inhibits host proteases that facilitate viral entry and possesses broad anti-inflammatory and immunomodulatory activities. Researchers have demonstrated that an interaction between SARS-CoV-2 spike protein (S) and lipopolysaccharides (LPS) enhances pro-inflammatory responses in vitro and in vivo. Hence, we wanted to understand the potential anti-inflammatory activities of plasma-derived and recombinant AAT (recAAT) in a model of human total peripheral blood mononuclear cells (PBMCs) exposed to a combination of CHO expressed trimeric spike protein and LPS, ex vivo. We confirmed that cytokine production was enhanced in PBMCs within six hours when low levels of LPS were combined with purified spike proteins (“spike”). In the presence of 0.5 mg/mL recAAT, however, LPS/spike-induced TNF-α and IL-1β mRNA expression and protein release were significantly inhibited (by about 46–50%) relative to LPS/spike alone. Although without statistical significance, recAAT also reduced production of IL-6 and IL-8. Notably, under the same experimental conditions, the plasma-derived AAT preparation Respreeza (used in native and oxidized forms) did not show significant effects. Our findings imply that an early pro-inflammatory activation of human PBMCs is better controlled by the recombinant version of AAT than the human plasma-derived AAT used here. Considering the increasing clinical interest in AAT therapy as useful to ameliorate the hyper-inflammation seen during COVID-19 infection, different AAT preparations require careful evaluation.

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PGE1-Containing Protocols Generate Mature (Leukemia-Derived) Dendritic Cells Directly from Leukemic Whole Blood

International Journal of Molecular Sciences ◽

10.3390/ijms20184590 ◽

2019 ◽

Vol 20 (18) ◽

pp. 4590

Author(s):

Daniel Christoph Amberger ◽

Fatemeh Doraneh-Gard ◽

Carina Gunsilius ◽

Melanie Weinmann ◽

Sabine Möbius ◽

...

Keyword(s):

Dendritic Cells ◽

Whole Blood ◽

Myeloid Leukemia ◽

Mononuclear Cells ◽

Ex Vivo ◽

Potential Treatment ◽

Cytokine Release ◽

Peripheral Blood Mononuclear ◽

Physiological Conditions ◽

Acute Myeloid

Dendritic cells (DCs) and leukemia-derived DC (DCleu) are potent stimulators of various immunoreactive cells and they play a pivotal role in the (re-) activation of the immune system. As a potential treatment tool for patients with acute myeloid leukemia, we developed and analyzed two new PGE1-containing protocols (Pici-PGE1, Kit M) to generate DC/DCleu ex vivo from leukemic peripheral blood mononuclear cells (PBMCs) or directly from leukemic whole blood (WB) to simulate physiological conditions. Pici-PGE1 generated significantly higher amounts of DCs from leukemic and healthy PBMCs when compared to control and comparable amounts as the already established protocol Pici-PGE2. The proportions of sufficient DC-generation were even higher after DC/DCleu-generation with Pici-PGE1. With Kits, it was possible to generate DCs and DCleu directly from leukemic and healthy WB without induction of blast proliferation. The average amounts of generated DCs and DCleu-subgroups were comparable with all Kits. The PGE1 containing Kit M generated significantly higher amounts of mature DCs when compared to the PGE2-containing Kit K and increased the anti-leukemic-activity. In summary PGE1-containing protocols were suitable for generating DC/DCleu from PBMCs as well as from WB, which reliably (re-) activated immunoreactive cells, improved the overall ex vivo anti-leukemic activity, and influenced cytokine-release-profiles.

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The Staphyloccous aureus Eap Protein Activates Expression of Proinflammatory Cytokines

Infection and Immunity ◽

10.1128/iai.01699-07 ◽

2008 ◽

Vol 76 (5) ◽

pp. 2164-2168 ◽

Cited By ~ 15

Author(s):

Thomas J. Scriba ◽

Sophie Sierro ◽

Eric L. Brown ◽

Rodney E. Phillips ◽

Andrew K. Sewell ◽

...

Keyword(s):

Proinflammatory Cytokines ◽

Mononuclear Cells ◽

Ex Vivo ◽

Human Peripheral Blood ◽

Intercellular Adhesion Molecule ◽

Intercellular Adhesion Molecule 1 ◽

Peripheral Blood Mononuclear ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Monocytes And Macrophages

ABSTRACT The extracellular adhesion protein (Eap) secreted by the major human pathogen Staphylococcus aureus is known to have several effects on human immunity. We have recently added to knowledge of these roles by demonstrating that Eap enhances interactions between major histocompatibility complex molecules and human leukocytes. Several studies have indicated that Eap can induce cytokine production by human peripheral blood mononuclear cells (PBMCs). To date, there has been no rigorous attempt to identify the breadth of cytokines produced by Eap stimulation or to identify the cell subsets that respond. Here, we demonstrate that Eap induces the secretion of the proinflammatory cytokines interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) by CD14+ leukocytes (monocytes and macrophages) within direct ex vivo PBMC populations (note that granulocytes are also CD14+ but are largely depleted from PBMC preparations). Anti-intercellular adhesion molecule 1 (CD54) antibodies inhibited this induction and implicated a role for this known Eap binding protein in cellular activation. IL-6 and TNF-α secretion by murine cells exposed to Eap was also observed. The activation of CD14+ cells by Eap suggests that it could play a significant role in both septic shock and fever, two of the major pathological features of S. aureus infections.

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MiR-144-3p is associated with pathological inflammation in patients infected with Mycobacteroides abscessus

Experimental & Molecular Medicine ◽

10.1038/s12276-020-00552-0 ◽

2021 ◽

Vol 53 (1) ◽

pp. 136-149

Author(s):

Hyeon Ji Kim ◽

In Soo Kim ◽

Sung-Gwon Lee ◽

Young Jae Kim ◽

Prashanta Silwal ◽

...

Keyword(s):

Proinflammatory Cytokines ◽

Mononuclear Cells ◽

Nontuberculous Mycobacteria ◽

Expression Profiles ◽

Host Factors ◽

Integrated Analysis ◽

Peripheral Blood Mononuclear ◽

Global Health Issue ◽

Chemokine Ligand ◽

Chemokine Ligand 2

AbstractInfection with rapidly growing nontuberculous mycobacteria is emerging as a global health issue; however, key host factors remain elusive. Here, we investigated the characteristic immune profiles of peripheral blood mononuclear cells (PBMCs) from patients infected with Mycobacteroides abscessus subsp. abscessus (Mabc) and M. abscessus subsp. massiliense (Mmass). Using an integrated analysis of global mRNA and microRNA expression profiles, we found that several inflammatory cytokines/chemokines [interleukin (IL)-1β, IL-6, C-X-C motif chemokine ligand 2, and C-C motif chemokine ligand 2] and miR-144-3p were significantly upregulated in PBMCs from patients compared with those from healthy controls (HCs). Notably, there was a strong correlation between the expression levels of miR-144-3p and proinflammatory cytokines/chemokines. Similarly, upregulated expression of miR-144-3p and proinflammatory cytokines/chemokines was found in macrophages and lungs from mice after infection with Mabc and Mmass. We showed that the expression of negative regulators of inflammation (SARM1 and TNIP3) was significantly downregulated in PBMCs from the patients, although they were not putative targets of miR-144-3p. Furthermore, overexpression of miR-144-3p led to a marked increase in proinflammatory cytokines/chemokines and promoted bacterial growth in macrophages. Together, our results highlight the importance of miR-144-3p linking to pathological inflammation during M. abscessus infection.

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Initiating antiretroviral treatment early in infancy has long-term benefits on the HIV reservoir in late childhood and adolescence

Clinical Infectious Diseases ◽

10.1093/cid/ciaa1931 ◽

2021 ◽

Author(s):

Véronique Avettand-Fenoel ◽

Jérôme Lechenadec ◽

Mariama Sadjo Diallo ◽

Marine Fillion ◽

Adeline Melard ◽

...

Keyword(s):

Mononuclear Cells ◽

Ex Vivo ◽

T Cell Subsets ◽

Effector Memory ◽

Childhood And Adolescence ◽

Older Children ◽

Peripheral Blood Mononuclear ◽

Hiv Reservoir ◽

Hiv Dna ◽

Cart Initiation

Abstract Background Early combined antiretroviral therapy (cART) limits the total HIV-DNA load in children. However, data on its impact in older children and adolescents remain scarce. This study aims to compare HIV reservoirs in children (5-12 years) and adolescents (13-17 years) who started cART before 6 months (early (E-)group) or after 2 years old (late (L-)group). Methods The ANRS-EP59-CLEAC study prospectively enrolled 76 HIV-1 perinatally-infected patients who reached HIV-RNA<400 copies/mL less than 24 months after cART initiation, regardless of subsequent viral suppression (E-group: 27 children, 9 adolescents; L-group: 19 children, 21 adolescents). Total and integrated HIV-DNA were quantified in blood and in CD4+ T cell subsets. A substudy assessed HIV reservoir inducibility after ex vivo peripheral blood mononuclear cells (PBMCs) stimulation. Results Total HIV-DNA levels were lower in early- than late-treated patients (Children: 2.14 vs 2.87 log cp/million PBMCs, p<0.0001; Adolescents: 2.25 vs 2.74log, p<0.0001). Low reservoir was independently associated with treatment precocity, protective HLA and low cumulative viremia since cART initiation. The 60 participants with undetectable integrated HIV-DNA started cART earlier than the other patients (4 vs 54 months, p=0.03). In those with sustained virological control, transitional memory and effector memory CD4+T cells were less infected in the E-group than in the L-group (p=0.03 and 0.02, respectively). Viral inducibility of reservoir cells after normalization to HIV-DNA levels was similar between the groups. Conclusions Early cART results in a smaller blood HIV reservoir until adolescence, but all tested participants had an inducible reservoir. This deserves cautious consideration for HIV remission strategies.

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Immunoglobulin A1 Protease, an Exoenzyme of Pathogenic Neisseriae, Is a Potent Inducer of Proinflammatory Cytokines

Journal of Experimental Medicine ◽

10.1084/jem.190.8.1049 ◽

1999 ◽

Vol 190 (8) ◽

pp. 1049-1058 ◽

Cited By ~ 52

Author(s):

Dirk R. Lorenzen ◽

Frank Düx ◽

Uwe Wölk ◽

Anastasios Tsirpouchtsidis ◽

Gaby Haas ◽

...

Keyword(s):

Proinflammatory Cytokines ◽

Mononuclear Cells ◽

Native Form ◽

Peripheral Blood Mononuclear ◽

Potent Inducer ◽

Cytokine Induction ◽

Regulatory Cytokine ◽

Iga1 Protease ◽

Tnf Α ◽

Necrosis Factor

A characteristic of human pathogenic Neisseriae is the production and secretion of an immunoglobulin (Ig)A1-specific serine protease (IgA1 protease) that cleaves preferentially human IgA1 and other target proteins. Here we show a novel function for native IgA1 protease, i.e., the induction of proinflammatory cytokines such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-8 from peripheral blood mononuclear cells. The capacity of IgA1 protease to elicit such cytokine responses in monocytes was enhanced in the presence of T lymphocytes. IgA1 protease did not induce the regulatory cytokine IL-10, which was, however, found in response to lipopolysaccharide and phytohemagglutinin. The immunomodulatory effects caused by IgA1 protease require a native form of the enzyme, and denaturation abolished cytokine induction. However, the proteolytic activity is not required for the cytokine induction by IgA1 protease. Our results indicate that IgA1 protease exhibits important immunostimulatory properties and may contribute substantially to the pathogenesis of neisserial infections by inducing large amounts of TNF-α and other proinflammatory cytokines. In particular, IgA1 protease may represent a key virulence determinant of bacterial meningitis.

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