interleukin-8 has been researched along with Coronavirus-Infections* in 32 studies
1 review(s) available for interleukin-8 and Coronavirus-Infections
1 trial(s) available for interleukin-8 and Coronavirus-Infections
31 other study(ies) available for interleukin-8 and Coronavirus-Infections
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Activation of the MKK3-p38-MK2-ZFP36 Axis by Coronavirus Infection Restricts the Upregulation of AU-Rich Element-Containing Transcripts in Proinflammatory Responses.
Coronavirus infections induce the expression of multiple proinflammatory cytokines and chemokines. We have previously shown that in cells infected with gammacoronavirus infectious bronchitis virus (IBV), interleukin 6 (IL-6), and IL-8 were drastically upregulated, and the MAP kinase p38 and the integrated stress response pathways were implicated in this process. In this study, we report that coronavirus infection activates a negative regulatory loop that restricts the upregulation of a number of proinflammatory genes. As revealed by the initial transcriptomic and subsequent validation analyses, the anti-inflammatory adenine-uridine (AU)-rich element (ARE)-binding protein, zinc finger protein 36 (ZFP36), and its related family members were upregulated in cells infected with IBV and three other coronaviruses, alphacoronaviruses porcine epidemic diarrhea virus (PEDV), human coronavirus 229E (HCoV-229E), and betacoronavirus HCoV-OC43, respectively. Characterization of the functional roles of ZFP36 during IBV infection demonstrated that ZFP36 promoted the degradation of transcripts coding for IL-6, IL-8, dual-specificity phosphatase 1 (DUSP1), prostaglandin-endoperoxide synthase 2 (PTGS2) and TNF-α-induced protein 3 (TNFAIP3), through binding to AREs in these transcripts. Consistently, knockdown and inhibition of JNK and p38 kinase activities reduced the expression of ZFP36, as well as the expression of IL-6 and IL-8. On the contrary, overexpression of mitogen-activated protein kinase kinase 3 (MKK3) and MAPKAP kinase-2 (MK2), the upstream and downstream kinases of p38, respectively, increased the expression of ZFP36 and decreased the expression of IL-8. Taken together, this study reveals an important regulatory role of the MKK3-p38-MK2-ZFP36 axis in coronavirus infection-induced proinflammatory response. Topics: Adenine; Animals; Cell Line; Chlorocebus aethiops; Coronavirus Infections; Gene Expression Regulation; Humans; Infectious bronchitis virus; Interleukin-6; Interleukin-8; Intracellular Signaling Peptides and Proteins; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Protein Serine-Threonine Kinases; Transcriptional Activation; Tristetraprolin; Up-Regulation; Uridine; Vero Cells | 2022 |
Chemokines induced by PEDV infection and chemotactic effects on monocyte, T and B cells.
Porcine epidemic diarrhea virus (PEDV) is a re-emerging pathogen that causes severe economic loss in the pig industry. The host's innate immune system is the first line of defense on virus invasion of the small intestinal epithelial cells. Chemokines, as a part of the innate immune system, play an important role in host immunity against infection, however, and their expression and chemotactic effect on key immune cells in PEDV infection remains unclear. In this study, cDNA microarray was firstly performed to analyzed ileum tissue of piglets on the third day after PEDV infection. The differentially expressed genes mainly involved in multiple biological processes, chemokine signaling pathway and cytokine receptor interaction signaling pathway had the highest enrichment according to GO and KEGG enrichment analysis. The expression levels of chemokines MCP-1, MIP-1β, IL-8, CXCL9, CXCL10 and CXCL13 in ileum of PEDV- infected piglets were significantly higher than those in the control group. The expression of chemokines in vivo experiment was further verified by RT-qPCR and ELISA using PEDV-infected IPEC-J2 cells. The results showed that the PEDV-infected IPEC-J2 cells had significantly induced protein expression of MCP-1, MIP-1β, IL-8, CXCL9, CXCL-10 and CXCL13. These results indicated that the changes of chemokines expressed in the ileum of piglets (in vivo) were consistent with those in IPEC-J2 cells (in vitro) after PEDV infection. Finally, the role of chemokines in immune cell migration during PEDV infection was illustrated by siRNA-mediated knock down method and the co-culture model of IPEC-J2 cells with peripheral blood leukocyte cells (PBLCs). The FACS analysis showed that MCP-1 induced by PEDV infection played a chemotactic effect on CD14 Topics: Animals; Cell Line; Chemokine CCL4; Coronavirus Infections; Interleukin-8; Monocytes; Porcine epidemic diarrhea virus; Swine; Swine Diseases | 2022 |
Screening of Lactic Acid Bacterial Strains with Antiviral Activity Against Porcine Epidemic Diarrhea.
Newly emerging and re-emerging viral infectious diseases cause significant economic losses in swine production. Efficacious vaccines have not yet been developed for several major swine infectious diseases, including porcine epidemic diarrhea virus (PEDV). We used the PEDV-infected Vero cell model to screen lactic acid bacteria (LAB) strains with antiviral activity. Sixty LAB strains were isolated from the feces of nursing piglets. After the elimination of LAB strains with high cytotoxicity to Vero cells, the protective effects of the remaining 6 strains against PEDV infection were determined. Vero cells pretreated with the intracellular extracts or cell wall fractions of YM22 and YM33 strains for 24 h before infection with PEDV showed significantly higher cell viabilities and lower mRNA expression of PEDV nucleocapsid (PEDV-N) than the unpretreated cells, indicating that the intracellular extracts and cell wall fractions of YM22 and YM33 possessed prophylactic effects on Vero cells against PEDV infection. PEDV-infection significantly increased the mRNA expression of proinflammatory cytokines, including tumor necrosis factor-α (TNF-α) and interleukin-8 (IL-8) in Vero cells. However, pretreatment of Vero cells with the cell wall fractions of YM22 and YM33 decreased the mRNA expression of TNF-α and IL-8, which could be a mechanism associated with the protective effects of YM22 and YM33 against PEDV. Based on the biochemical characteristics and phylogenetic analyses, YM22 and YM33 were identified as Ligilactobacillus agilis (basonym: Lactobacillus agilis) and Ligilactobacillus salivarius (basonym: Lactobacillus salivarius), respectively. These findings suggest that L. agilis YM22 and L. salivarius YM33 could provide some levels of protective effects against PEDV infections. Topics: Animals; Antiviral Agents; Chlorocebus aethiops; Coronavirus Infections; Diarrhea; Dysentery; Interleukin-8; Lactic Acid; Lactobacillales; Phylogeny; Plant Extracts; Porcine epidemic diarrhea virus; RNA, Messenger; Swine; Swine Diseases; Tumor Necrosis Factor-alpha; Vero Cells | 2022 |
Porcine Hemagglutinating Encephalomyelitis Virus Infection
Porcine hemagglutinating encephalomyelitis virus (PHEV) is a betacoronavirus that causes vomiting and wasting disease and/or encephalomyelitis in suckling pigs. This study characterized PHEV infection, pathogenesis, and immune response in cesarean-derived, colostrum-deprived (CDCD) neonatal pigs. Infected animals developed mild respiratory, enteric, and neurological clinical signs between 2 to 13 days postoronasal inoculation (dpi). PHEV did not produce viremia, but virus shedding was detected in nasal secretions (1 to 10 dpi) and feces (2 to 7 dpi) by reverse transcriptase quantitative PCR (RT-qPCR). Viral RNA was detected in all tissues except liver, but the detection rate and RT-qPCR threshold cycle ( Topics: Animals; Betacoronavirus 1; Cell Line; Coronavirus Infections; Interferon-alpha; Interleukin-8; Organ Specificity; Reverse Transcriptase Polymerase Chain Reaction; Swine; Swine Diseases; T-Lymphocytes | 2021 |
Induction of the Proinflammatory Chemokine Interleukin-8 Is Regulated by Integrated Stress Response and AP-1 Family Proteins Activated during Coronavirus Infection.
Infection induces the production of proinflammatory cytokines and chemokines such as interleukin-8 (IL-8) and IL-6. Although they facilitate local antiviral immunity, their excessive release leads to life-threatening cytokine release syndrome, exemplified by the severe cases of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. In this study, we investigated the roles of the integrated stress response (ISR) and activator protein-1 (AP-1) family proteins in regulating coronavirus-induced IL-8 and IL-6 upregulation. The mRNA expression of IL-8 and IL-6 was significantly induced in cells infected with infectious bronchitis virus (IBV), a gammacoronavirus, and porcine epidemic diarrhea virus, an alphacoronavirus. Overexpression of a constitutively active phosphomimetic mutant of eukaryotic translation initiation factor 2α (eIF2α), chemical inhibition of its dephosphorylation, or overexpression of its upstream double-stranded RNA-dependent protein kinase (PKR) significantly enhanced IL-8 mRNA expression in IBV-infected cells. Overexpression of the AP-1 protein cJUN or its upstream kinase also increased the IBV-induced IL-8 mRNA expression, which was synergistically enhanced by overexpression of cFOS. Taken together, this study demonstrated the important regulatory roles of ISR and AP-1 proteins in IL-8 production during coronavirus infection, highlighting the complex interactions between cellular stress pathways and the innate immune response. Topics: Alphacoronavirus; Animals; Cell Line; Chlorocebus aethiops; Coronavirus Infections; eIF-2 Kinase; Endoplasmic Reticulum Stress; Eukaryotic Initiation Factor-2; Gammacoronavirus; Gene Expression Regulation; Humans; Immunity, Innate; Infectious bronchitis virus; Interleukin-8; Phosphorylation; Porcine epidemic diarrhea virus; Proto-Oncogene Proteins c-fos; Proto-Oncogene Proteins c-jun; Signal Transduction; Transcription Factor AP-1; Unfolded Protein Response; Up-Regulation; Vero Cells | 2021 |
Elevated Expression Levels of Lung Complement Anaphylatoxin, Neutrophil Chemoattractant Chemokine IL-8, and RANTES in MERS-CoV-Infected Patients: Predictive Biomarkers for Disease Severity and Mortality.
The complement system, a network of highly-regulated proteins, represents a vital part of the innate immune response. Over-activation of the complement system plays an important role in inflammation, tissue damage, and infectious disease severity. The prevalence of MERS-CoV in Saudi Arabia remains significant and cases are still being reported. The role of complement in Middle East Respiratory Syndrome coronavirus (MERS-CoV) pathogenesis and complement-modulating treatment strategies has received limited attention, and studies involving MERS-CoV-infected patients have not been reported. This study offers the first insight into the pulmonary expression profile including seven complement proteins, complement regulatory factors, IL-8, and RANTES in MERS-CoV infected patients without underlying chronic medical conditions. Our results significantly indicate high expression levels of complement anaphylatoxins (C3a and C5a), IL-8, and RANTES in the lungs of MERS-CoV-infected patients. The upregulation of lung complement anaphylatoxins, C5a, and C3a was positively correlated with IL-8, RANTES, and the fatality rate. Our results also showed upregulation of the positive regulatory complement factor P, suggesting positive regulation of the complement during MERS-CoV infection. High levels of lung C5a, C3a, factor P, IL-8, and RANTES may contribute to the immunopathology, disease severity, ARDS development, and a higher fatality rate in MERS-CoV-infected patients. These findings highlight the potential prognostic utility of C5a, C3a, IL-8, and RANTES as biomarkers for MERS-CoV disease severity and mortality. To further explore the prediction of functional partners (proteins) of highly expressed proteins (C5a, C3a, factor P, IL-8, and RANTES), the computational protein-protein interaction (PPI) network was constructed, and six proteins (hub nodes) were identified. Topics: Aged; Biomarkers; Chemokine CCL5; Complement C3a; Complement C5a; Coronavirus Infections; Female; Humans; Interleukin-8; Lung; Male; Middle Aged; Middle East Respiratory Syndrome Coronavirus; Prognosis; Severity of Illness Index; Survival Analysis; Up-Regulation | 2021 |
Threatening drug-drug interaction in a kidney transplant patient with coronavirus disease 2019 (COVID-19).
During the novel coronavirus pandemic, organ transplant recipients represent a frail susceptible category due to long-term immunosuppressive therapy. For this reason, clinical manifestations may differ from general population and different treatment approaches may be needed. We present the case of a 36-year-old kidney-transplanted woman affected by Senior-Loken syndrome diagnosed with COVID-19 pneumonia after a contact with her positive mother. Initial symptoms were fatigue, dry cough, and coryza; she never had fever nor oxygen supplementation. Hydroxychloroquine and lopinavir/ritonavir were started, and the antiviral drug was replaced with darunavir/cobicistat after 2 days for diarrhea. Immunosuppressant levels were closely monitored, and we observed very high tacrolimus trough levels despite initial dose reduction. The patient was left with steroid therapy alone. The peculiarity of clinical presentation and the management difficulties represent the flagship of our case report. We stress the need for guidelines in transplant recipients with COVID-19 infection with particular regard to the management of therapy. Topics: Adult; Antiviral Agents; Betacoronavirus; C-Reactive Protein; Ciliopathies; Cobicistat; Common Cold; Coronavirus Infections; Cough; COVID-19; COVID-19 Drug Treatment; Cytochrome P-450 CYP3A Inhibitors; Darunavir; Deprescriptions; Drug Combinations; Drug Interactions; Enzyme Inhibitors; Fatigue; Female; Glucocorticoids; Graft Rejection; Humans; Hydroxychloroquine; Immunocompromised Host; Immunosuppressive Agents; Interleukin-10; Interleukin-1beta; Interleukin-6; Interleukin-8; Kidney Diseases, Cystic; Kidney Failure, Chronic; Kidney Transplantation; Leber Congenital Amaurosis; Lopinavir; Methylprednisolone; Optic Atrophies, Hereditary; Pandemics; Pneumonia, Viral; Ritonavir; SARS-CoV-2; Severity of Illness Index; Tacrolimus | 2020 |
Dapsone, colchicine and olanzapine as treatment adjuncts to prevent COVID-19 associated adult respiratory distress syndrome (ARDS).
Topics: Adult; Betacoronavirus; Chemotaxis, Leukocyte; Colchicine; Coronavirus Infections; COVID-19; Dapsone; Humans; Interleukin-8; Neutrophils; Olanzapine; Pandemics; Pneumonia, Viral; Respiratory Distress Syndrome; SARS-CoV-2 | 2020 |
Using IL-2R/lymphocytes for predicting the clinical progression of patients with COVID-19.
Effective laboratory markers for the estimation of disease severity and predicting the clinical progression of coronavirus disease-2019 (COVID-19) is urgently needed. Laboratory tests, including blood routine, cytokine profiles and infection markers, were collected from 389 confirmed COVID-19 patients. The included patients were classified into mild (n = 168), severe (n = 169) and critical groups (n = 52). The leukocytes, neutrophils, infection biomarkers [such as C-reactive protein (CRP), procalcitonin (PCT) and ferritin] and the concentrations of cytokines [interleukin (IL)-2R, IL-6, IL-8, IL-10 and tumor necrosis factor (TNF)-α] were significantly increased, while lymphocytes were significantly decreased with increased severity of illness. The amount of IL-2R was positively correlated with the other cytokines and negatively correlated with lymphocyte number. The ratio of IL-2R to lymphocytes was found to be remarkably increased in severe and critical patients. IL-2R/lymphocytes were superior compared with other markers for the identification of COVID-19 with critical illness, not only from mild but also from severe illness. Moreover, the cytokine profiles and IL-2R/lymphocytes were significantly decreased in recovered patients, but further increased in disease-deteriorated patients, which might be correlated with the outcome of COVID-19. Lymphopenia and increased levels of cytokines were closely associated with disease severity. The IL-2R/lymphocyte was a prominent biomarker for early identification of severe COVID-19 and predicting the clinical progression of the disease. Topics: Aged; Aged, 80 and over; Betacoronavirus; Biomarkers; C-Reactive Protein; China; Coronavirus Infections; COVID-19; Disease Progression; Female; Ferritins; Humans; Interleukin-10; Interleukin-2 Receptor alpha Subunit; Interleukin-6; Interleukin-8; Leukocyte Count; Male; Middle Aged; Neutrophils; Pandemics; Pneumonia, Viral; Procalcitonin; Prognosis; SARS-CoV-2; Severity of Illness Index; T-Lymphocytes; Tumor Necrosis Factor-alpha | 2020 |
Steroid-Responsive Encephalitis in Coronavirus Disease 2019.
Coronavirus disease 2019 (COVID-19) infection has the potential for targeting the central nervous system, and several neurological symptoms have been described in patients with severe respiratory distress. Here, we described the case of a 60-year-old patient with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection but only mild respiratory abnormalities who developed an akinetic mutism attributable to encephalitis. Magnetic resonance imaging was negative, whereas electroencephalography showed generalized theta slowing. Cerebrospinal fluid analyses during the acute stage were negative for SARS-CoV-2, positive for pleocytosis and hyperproteinorrachia, and showed increased interleukin-8 and tumor necrosis factor-α concentrations. Other infectious or autoimmune disorders were excluded. A progressive clinical improvement along with a reduction of cerebrospinal fluid parameters was observed after high-dose steroid treatment, thus arguing for an inflammatory-mediated brain involvement related to COVID-19. ANN NEUROL 2020;88:423-427. Topics: Akinetic Mutism; Antiviral Agents; beta 2-Microglobulin; Betacoronavirus; Coronavirus Infections; COVID-19; COVID-19 Drug Treatment; Drug Combinations; Electroencephalography; Encephalitis; Glucocorticoids; Humans; Hydroxychloroquine; Interleukin-6; Interleukin-8; Lopinavir; Magnetic Resonance Imaging; Male; Methylprednisolone; Middle Aged; Pandemics; Pneumonia, Viral; Ritonavir; SARS-CoV-2; Treatment Outcome; Tumor Necrosis Factor-alpha | 2020 |
Viral and host factors related to the clinical outcome of COVID-19.
In December 2019, coronavirus disease 2019 (COVID-19), which is caused by the new coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), was identified in Wuhan (Hubei province, China) Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aging; Animals; Asymptomatic Infections; Betacoronavirus; China; Cohort Studies; Coronavirus Infections; COVID-19; Critical Illness; Disease Progression; Evolution, Molecular; Female; Genetic Variation; Genome, Viral; Hospitalization; Host-Pathogen Interactions; Humans; Inflammation Mediators; Interleukin-6; Interleukin-8; Lymphocyte Count; Lymphopenia; Male; Middle Aged; Pandemics; Phylogeny; Pneumonia, Viral; Respiratory Distress Syndrome; SARS-CoV-2; T-Lymphocytes; Time Factors; Treatment Outcome; Virulence; Virus Shedding; Young Adult; Zoonoses | 2020 |
Pediatric Crohn Disease and Multisystem Inflammatory Syndrome in Children (MIS-C) and COVID-19 Treated With Infliximab.
Coronavirus disease 2019 (COVID-19) may lead to a severe inflammatory response referred to as a cytokine storm. We describe a case of severe COVID-19 infection in a recently diagnosed pediatric Crohn disease patient successfully treated with tumor necrosis factor-alpha (TNF-α) blockade. The patient presented with 5 days of fever, an erythematous maculopapular facial rash, and abdominal pain without respiratory symptoms. SARS-CoV-2 polymerase chain reaction was positive. Despite inpatient treatment for COVID-19 and a perianal abscess, the patient acutely decompensated, with worsening fever, tachycardia, fluid-refractory hypotension, elevation of liver enzymes, and transformation of the rash into purpura extending from the face to the trunk, upper and lower extremities, including the palmar and plantar surfaces of the hands and feet. Cytokine profile revealed rising levels of interleukin (IL)-6, IL-8, and TNF-α, higher than those described in either inflammatory bowel disease or severe COVID-19 alone. The patient was treated with infliximab for TNF-α blockade to address both moderately to severely active Crohn disease and multisystem inflammatory syndrome in children temporally related to COVID-19. Within hours of infliximab treatment, fever, tachycardia, and hypotension resolved. Cytokine profile improved with normalization of TNF-α, a decrease in IL-6, and IL-8 concentrations. This case supports a role for blockade of TNF-α in the treatment of COVID-19 inflammatory cascade. The role of anti-TNF agents in patients with multisystem inflammatory syndrome in children temporally related to COVID-19 requires further investigation. Topics: Abnormalities, Multiple; Adolescent; Antirheumatic Agents; Betacoronavirus; Coronavirus Infections; COVID-19; Crohn Disease; Genetic Diseases, X-Linked; Humans; Ichthyosiform Erythroderma, Congenital; Infliximab; Interleukin-6; Interleukin-8; Limb Deformities, Congenital; Male; Pandemics; Pneumonia, Viral; SARS-CoV-2; Tumor Necrosis Factor-alpha | 2020 |
IL-6 and CD8+ T cell counts combined are an early predictor of in-hospital mortality of patients with COVID-19.
BACKGROUNDFatal cases of COVID-19 are increasing globally. We retrospectively investigated the potential of immunologic parameters as early predictors of COVID-19.METHODSA total of 1018 patients with confirmed COVID-19 were enrolled in our 2-center retrospective study. Clinical feature, laboratory test, immunological test, radiological findings, and outcomes data were collected. Univariate and multivariable logistic regression analyses were performed to evaluate factors associated with in-hospital mortality. Receiver operator characteristic (ROC) curves and survival curves were plotted to evaluate their clinical utility.RESULTSThe counts of all T lymphocyte subsets were markedly lower in nonsurvivors than in survivors, especially CD8+ T cells. Among all tested cytokines, IL-6 was elevated most significantly, with an upward trend of more than 10-fold. Using multivariate logistic regression analysis, IL-6 levels of more than 20 pg/mL and CD8+ T cell counts of less than 165 cells/μL were found to be associated with in-hospital mortality after adjusting for confounding factors. Groups with IL-6 levels of more than 20 pg/mL and CD8+ T cell counts of less than 165 cells/μL had a higher percentage of older and male patients as well as a higher proportion of patients with comorbidities, ventilation, intensive care unit admission, shock, and death. Furthermore, the receiver operating curve of the model combining IL-6 (>20 pg/mL) and CD8+ T cell counts (<165 cells/μL) displayed a more favorable discrimination than that of the CURB-65 score. The Hosmer-Lemeshow test showed a good fit of the model, with no statistical significance.CONCLUSIONIL-6 (>20 pg/mL) and CD8+ T cell counts (<165 cells/μL) are 2 reliable prognostic indicators that accurately stratify patients into risk categories and predict COVID-19 mortality.FundingThis work was supported by funding from the National Natural Science Foundation of China (no. 81772477 and 81201848). Topics: Aged; Area Under Curve; Betacoronavirus; CD8-Positive T-Lymphocytes; Coronavirus Infections; COVID-19; Female; Hospital Mortality; Humans; Interleukin-10; Interleukin-6; Interleukin-8; Logistic Models; Lymphocyte Count; Lymphopenia; Male; Middle Aged; Multivariate Analysis; Pandemics; Pneumonia, Viral; Prognosis; Receptors, Interleukin-2; Retrospective Studies; ROC Curve; SARS-CoV-2; Tumor Necrosis Factor-alpha | 2020 |
Characterization of the Inflammatory Response to Severe COVID-19 Illness.
Topics: Acute-Phase Reaction; Adult; Aged; alpha 1-Antitrypsin; Betacoronavirus; Blotting, Western; Carrier Proteins; Case-Control Studies; Community-Acquired Infections; Coronavirus Infections; COVID-19; Critical Illness; Cytokines; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Female; Hospitalization; Humans; Hypoxia-Inducible Factor 1, alpha Subunit; Intensive Care Units; Interleukin-10; Interleukin-1beta; Interleukin-6; Interleukin-8; Lactic Acid; Length of Stay; Male; Membrane Proteins; Middle Aged; Neutrophils; Pandemics; Phosphorylation; Pneumonia; Pneumonia, Viral; Receptors, Tumor Necrosis Factor, Type I; SARS-CoV-2; Severity of Illness Index; Thyroid Hormone-Binding Proteins; Thyroid Hormones | 2020 |
Use of inhaled corticosteroids in asthma and coronavirus disease 2019: Keep calm and carry on.
Topics: Administration, Inhalation; Adrenal Cortex Hormones; Angiotensin-Converting Enzyme 2; Anti-Asthmatic Agents; Asthma; Betacoronavirus; Coronavirus Infections; COVID-19; Cytokine Release Syndrome; Dexamethasone; Endoribonucleases; Gene Expression Regulation; Host-Pathogen Interactions; Humans; Interleukin-6; Interleukin-8; Pandemics; Peptidyl-Dipeptidase A; Pneumonia, Viral; Receptors, Virus; SARS-CoV-2; Serine Endopeptidases; Severity of Illness Index; Tumor Necrosis Factor-alpha; Viral Nonstructural Proteins | 2020 |
Gene expression pattern differences in primary human pulmonary epithelial cells infected with MERS-CoV or SARS-CoV-2.
Coronaviruses such as MERS-CoV and SARS-CoV-2 infect the human respiratory tract and can cause severe pneumonia. Disease severity and outcomes are different for these two infections: the human mortality rate for MERS-CoV and SARS-CoV-2 is over 30% and less than 10%, respectively. Here, using microarray assay, we analyzed the global alterations in gene expression induced by MERS-CoV or SARS-CoV-2 infections in primary human pulmonary epithelial cells. Overall, the number of differentially expressed genes was higher in human lung cells infected with MERS-CoV than in cells with SARS-CoV-2. Out of 44,556 genes analyzed, 127 and 50 were differentially expressed in cells infected with MERS-CoV and SARS-CoV-2, respectively (> 2-fold increase, compared to uninfected cells). Of these, only eight genes, including the one coding for CXCL8, were similarly modulated (upregulated or downregulated) by the two coronaviruses. Importantly, these results were virus-specific and not conditioned by differences in viral load, and viral growth curves were similar in human lung cells infected with both viruses. Our results suggest that these distinct gene expression profiles, detected early after infection by these two coronaviruses, may help us understand the differences in clinical outcomes of MERS-CoV and SARS-CoV-2 infections. Topics: Betacoronavirus; Cells, Cultured; Chemokine CXCL6; Coronavirus Infections; COVID-19; Down-Regulation; Epithelial Cells; Gene Expression Profiling; Host Microbial Interactions; Humans; Interleukin-8; Lung; Middle East Respiratory Syndrome Coronavirus; Pandemics; Pneumonia, Viral; SARS-CoV-2; Species Specificity; Up-Regulation | 2020 |
Cytokine prediction of mortality in COVID19 patients.
Coronavirus disease 2019 (COVID19) is a life-threatening infection with uncertain progression and outcome. Assessing the severity of the disease for worsening patients is of importance in making decisions related to supportive mechanical ventilation and aggressive treatments. This was a prospective, non-randomized study that included hospitalized patients diagnosed with COVID19. Pro-inflammatory cytokines were assessed during hospitalization, and we calculated a prediction paradigm for 30-day mortality based on the serum levels of interleukin1β (IL1β), interleukin6 (IL6), interleukin8 (IL8), and tumor necrosis factor alpha (TNFα) measured by next-generation ELISA. Data of 71 COVID19 patients, mean age 62 years, SD13.8, 50 males, 21 females, were analyzed. Twelve (16.9%) patients died within 7-39 days of their first COVID19 positive nasopharyngeal test. Levels of IL6 and TNFα were significantly higher in patients that did not survive. IL6 predicted mortality at the cut-off value of 163.4 pg/ml, with a sensitivity of 91.7% and specificity of 57.6%. Our findings demonstrate that IL6 expression is significant for the prediction of 30-day mortality in hospitalized COVID19 patients and, therefore, may assist in treatment decisions. Topics: Betacoronavirus; Coronavirus Infections; COVID-19; Cytokine Release Syndrome; Enzyme-Linked Immunosorbent Assay; Female; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Kaplan-Meier Estimate; Male; Middle Aged; Pandemics; Pneumonia, Viral; Prognosis; Prospective Studies; SARS-CoV-2; Tumor Necrosis Factor-alpha | 2020 |
Cytokine profile in plasma of severe COVID-19 does not differ from ARDS and sepsis.
BACKGROUNDElevated levels of inflammatory cytokines have been associated with poor outcomes among COVID-19 patients. It is unknown, however, how these levels compare with those observed in critically ill patients with acute respiratory distress syndrome (ARDS) or sepsis due to other causes.METHODSWe used a Luminex assay to determine expression of 76 cytokines from plasma of hospitalized COVID-19 patients and banked plasma samples from ARDS and sepsis patients. Our analysis focused on detecting statistical differences in levels of 6 cytokines associated with cytokine storm (IL-1β, IL-1RA, IL-6, IL-8, IL-18, and TNF-α) between patients with moderate COVID-19, severe COVID-19, and ARDS or sepsis.RESULTSFifteen hospitalized COVID-19 patients, 9 of whom were critically ill, were compared with critically ill patients with ARDS (n = 12) or sepsis (n = 16). There were no statistically significant differences in baseline levels of IL-1β, IL-1RA, IL-6, IL-8, IL-18, and TNF-α between patients with COVID-19 and critically ill controls with ARDS or sepsis.CONCLUSIONLevels of inflammatory cytokines were not higher in severe COVID-19 patients than in moderate COVID-19 or critically ill patients with ARDS or sepsis in this small cohort. Broad use of immunosuppressive therapies in ARDS has failed in numerous Phase 3 studies; use of these therapies in unselected patients with COVID-19 may be unwarranted.FUNDINGFunding was received from NHLBI K23 HL125663 (AJR); The Bill and Melinda Gates Foundation OPP1113682 (AJR and CAB); Burroughs Wellcome Fund Investigators in the Pathogenesis of Infectious Diseases #1016687 NIH/NIAID U19AI057229-16; Stanford Maternal Child Health Research Institute; and Chan Zuckerberg Biohub (CAB). Topics: Adult; Aged; Case-Control Studies; Coronavirus Infections; COVID-19; Cytokine Release Syndrome; Cytokines; Female; Humans; Interleukin 1 Receptor Antagonist Protein; Interleukin-18; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Middle Aged; Pandemics; Pneumonia, Viral; Respiratory Distress Syndrome; Sepsis; Severity of Illness Index; Tumor Necrosis Factor-alpha | 2020 |
Tobacco, but Not Nicotine and Flavor-Less Electronic Cigarettes, Induces ACE2 and Immune Dysregulation.
The COVID-19 pandemic caused by the SARS-CoV-2 virus, overlaps with the ongoing epidemics of cigarette smoking and electronic cigarette (e-cig) vaping. However, there is scarce data relating COVID-19 risks and outcome with cigarette or e-cig use. In this study, we mined three independent RNA expression datasets from smokers and vapers to understand the potential relationship between vaping/smoking and the dysregulation of key genes and pathways related to COVID-19. We found that smoking, but not vaping, upregulates ACE2, the cellular receptor that SARS-CoV-2 requires for infection. Both smoking and use of nicotine and flavor-containing e-cigs led to upregulation of pro-inflammatory cytokines and inflammasome-related genes. Specifically, chemokines including CCL20 and CXCL8 are upregulated in smokers, and CCL5 and CCR1 are upregulated in flavor/nicotine-containing e-cig users. We also found genes implicated in inflammasomes, such as CXCL1, CXCL2, NOD2, and ASC, to be upregulated in smokers and these e-cig users. Vaping flavor and nicotine-less e-cigs, however, did not lead to significant cytokine dysregulation and inflammasome activation. Release of inflammasome products, such as IL-1B, and cytokine storms are hallmarks of COVID-19 infection, especially in severe cases. Therefore, our findings demonstrated that smoking or vaping may critically exacerbate COVID-19-related inflammation or increase susceptibility to COVID-19. Topics: Adult; Angiotensin-Converting Enzyme 2; Betacoronavirus; Bronchi; Chemokine CCL20; Coronavirus Infections; COVID-19; Electronic Nicotine Delivery Systems; Epithelial Cells; Humans; Immune System; Interleukin-1beta; Interleukin-8; Middle Aged; Nod2 Signaling Adaptor Protein; Pandemics; Peptidyl-Dipeptidase A; Pneumonia, Viral; SARS-CoV-2; Tobacco Smoking; Up-Regulation; Young Adult | 2020 |
An inflammatory cytokine signature predicts COVID-19 severity and survival.
Several studies have revealed that the hyper-inflammatory response induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a major cause of disease severity and death. However, predictive biomarkers of pathogenic inflammation to help guide targetable immune pathways are critically lacking. We implemented a rapid multiplex cytokine assay to measure serum interleukin (IL)-6, IL-8, tumor necrosis factor (TNF)-α and IL-1β in hospitalized patients with coronavirus disease 2019 (COVID-19) upon admission to the Mount Sinai Health System in New York. Patients (n = 1,484) were followed up to 41 d after admission (median, 8 d), and clinical information, laboratory test results and patient outcomes were collected. We found that high serum IL-6, IL-8 and TNF-α levels at the time of hospitalization were strong and independent predictors of patient survival (P < 0.0001, P = 0.0205 and P = 0.0140, respectively). Notably, when adjusting for disease severity, common laboratory inflammation markers, hypoxia and other vitals, demographics, and a range of comorbidities, IL-6 and TNF-α serum levels remained independent and significant predictors of disease severity and death. These findings were validated in a second cohort of patients (n = 231). We propose that serum IL-6 and TNF-α levels should be considered in the management and treatment of patients with COVID-19 to stratify prospective clinical trials, guide resource allocation and inform therapeutic options. Topics: Aged; Betacoronavirus; Coronavirus Infections; COVID-19; Cytokines; Female; Hospitalization; Humans; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Middle Aged; Pandemics; Pneumonia, Viral; SARS-CoV-2; Severity of Illness Index; Survival Rate; Tumor Necrosis Factor-alpha | 2020 |
Re-analysis of Single Cell Transcriptome Reveals That the NR3C1-CXCL8-Neutrophil Axis Determines the Severity of COVID-19.
SARS-CoV-2 infection has recently been declared a pandemic. Some patients showing severe symptoms exhibit drastic inflammation and airway damage. In this study, we re-analyzed published scRNA-seq data of COVID-19 patient bronchoalveolar lavage fluid to further classify and compare immunological features according to the patient's disease severity. Patients with severe symptoms showed DNA damage and apoptotic features of epithelial cells. Our results suggested that epithelial damage was associated with neutrophil infiltration. Myeloid cells of severe patients showed higher expression of proinflammatory cytokines and chemokines such as CXCL8. As a result, neutrophils were abundant in lungs of patients from the severe group. Furthermore, recruited neutrophils highly expressed genes related to neutrophil extracellular traps. Neutrophil-mediated inflammation was regulated by glucocorticoid receptor expression and activity. Based on these results, we suggest that severe COVID-19 symptoms may be determined by differential expression of glucocorticoid receptors and neutrophils. Topics: Adult; Aged; Betacoronavirus; Bronchoalveolar Lavage Fluid; Coronavirus Infections; COVID-19; Epithelial Cells; Extracellular Traps; Female; Gene Expression Profiling; Humans; Inflammation; Interleukin-8; Male; Middle Aged; Myeloid Cells; Neutrophil Infiltration; Neutrophils; Pandemics; Pneumonia, Viral; Receptors, Glucocorticoid; RNA-Seq; SARS-CoV-2; Severity of Illness Index; Single-Cell Analysis; Transcriptome | 2020 |
Immune characteristics distinguish patients with severe disease associated with SARS-CoV-2.
Topics: Adult; Aged; Betacoronavirus; Biomarkers; Coronavirus Infections; COVID-19; Cytokine Release Syndrome; Female; Humans; Interleukin-2 Receptor alpha Subunit; Interleukin-6; Interleukin-8; Length of Stay; Lymphocyte Count; Male; Middle Aged; Neutrophils; Pandemics; Pneumonia, Viral; Retrospective Studies; SARS-CoV-2; Severity of Illness Index; Treatment Outcome; Young Adult | 2020 |
SARS-CoV-2 triggers inflammatory responses and cell death through caspase-8 activation.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection can lead to respiratory illness and multi-organ failure in critically ill patients. Although the virus-induced lung damage and inflammatory cytokine storm are believed to be directly associated with coronavirus disease 2019 (COVID-19) clinical manifestations, the underlying mechanisms of virus-triggered inflammatory responses are currently unknown. Here we report that SARS-CoV-2 infection activates caspase-8 to trigger cell apoptosis and inflammatory cytokine processing in the lung epithelial cells. The processed inflammatory cytokines are released through the virus-induced necroptosis pathway. Virus-induced apoptosis, necroptosis, and inflammation activation were also observed in the lung sections of SARS-CoV-2-infected HFH4-hACE2 transgenic mouse model, a valid model for studying SARS-CoV-2 pathogenesis. Furthermore, analysis of the postmortem lung sections of fatal COVID-19 patients revealed not only apoptosis and necroptosis but also massive inflammatory cell infiltration, necrotic cell debris, and pulmonary interstitial fibrosis, typical of immune pathogenesis in the lung. The SARS-CoV-2 infection triggered a dual mode of cell death pathways and caspase-8-dependent inflammatory responses may lead to the lung damage in the COVID-19 patients. These discoveries might assist the development of therapeutic strategies to treat COVID-19. Topics: Animals; Apoptosis; Betacoronavirus; Caspase 8; Cell Line, Tumor; Chemokine CCL5; Chemokine CXCL10; Coronavirus Infections; COVID-19; Cytokine Release Syndrome; Disease Models, Animal; Epithelial Cells; Gene Expression Regulation; Humans; Interleukin-1beta; Interleukin-7; Interleukin-8; Lung; Mice; Mice, Transgenic; Necroptosis; Pandemics; Pneumonia, Viral; Pulmonary Fibrosis; SARS-CoV-2; Tumor Necrosis Factor-alpha | 2020 |
Age-severity matched cytokine profiling reveals specific signatures in Covid-19 patients.
A global effort is currently undertaken to restrain the COVID-19 pandemic. Host immunity has come out as a determinant for COVID-19 clinical outcomes, and several studies investigated the immune profiling of SARS-CoV-2 infected people to properly direct the clinical management of the disease. Thus, lymphopenia, T-cell exhaustion, and the increased levels of inflammatory mediators have been described in COVID-19 patients, in particular in severe cases Topics: Age Factors; Aged; Aged, 80 and over; Antibodies, Viral; Betacoronavirus; Cluster Analysis; Coronavirus Infections; COVID-19; Cytokines; Female; Humans; Immunoglobulin G; Interleukin-10; Interleukin-8; Length of Stay; Leukocytes, Mononuclear; Male; Middle Aged; Pandemics; Pneumonia, Viral; SARS-CoV-2; Severity of Illness Index; Tumor Necrosis Factor-alpha | 2020 |
Broncho-alveolar inflammation in COVID-19 patients: a correlation with clinical outcome.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic proportions. Given that the main target of SARS-CoV-2 are lungs leading to severe pneumonia with hyperactivation of the inflammatory cascade, we conducted a prospective study to assess alveolar inflammatory status in patients with moderate to severe COVID-19.. Diagnostic bronchoalveolar lavage (BAL) was performed in 33 adult patients with SARS-CoV-2 infection by real-time PCR on nasopharyngeal swab admitted to the Intensive care unit (ICU) (n = 28) and to the Intermediate Medicine Ward (IMW) (n = 5). We analyze the differential cell count, ultrastructure of cells and Interleukin (IL)6, 8 and 10 levels.. ICU patients showed a marked increase in neutrophils (1.24 × 10. Alveolitis, associated with COVID-19, is mainly sustained by innate effectors which showed features of extensive activation. The burden of pro-inflammatory cytokines IL6 and IL8 in the broncho-alveolar environment is associated with clinical outcome. Topics: Adenosine Monophosphate; Adrenal Cortex Hormones; Aged; Alanine; Antibodies, Monoclonal, Humanized; Antiviral Agents; Betacoronavirus; Bronchoalveolar Lavage; Bronchoalveolar Lavage Fluid; Coronavirus Infections; COVID-19; COVID-19 Drug Treatment; Drug Combinations; Female; Humans; Hydroxychloroquine; Inflammation; Intensive Care Units; Interleukin-10; Interleukin-6; Interleukin-8; Italy; Leukocytes; Leukocytes, Mononuclear; Lopinavir; Lung; Lymphocytes; Macrophages, Alveolar; Male; Microscopy, Electron, Transmission; Middle Aged; Neutrophils; Pandemics; Pneumonia, Viral; Prognosis; Prospective Studies; Respiration, Artificial; Ritonavir; SARS-CoV-2; Spike Glycoprotein, Coronavirus; Survival Rate; Virion | 2020 |
Porcine epidemic diarrhea virus nsp4 induces pro-inflammatory cytokine and chemokine expression inhibiting viral replication in vitro.
Porcine epidemic diarrhea virus (PEDV) causes severe economic loss in the pig industry each year. To better understand the relationship between cytokines and PEDV replication, in this study, pro-inflammatory cytokine and chemokine expression profiles in Vero cells infected with PEDV were analyzed. Real-time quantitative PCR assay indicated that IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 expression levels were significantly upregulated. Moreover, overexpression and siRNA silencing assays showed that overexpression of IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 could significantly inhibit PEDV replication, while silencing of IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 could significantly promote PEDV replication. Finally, a dual-luciferase reporter assay showed that nsp4 contributed to the expression of IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 via the NF-κB pathway. Together, these data determined that PEDV nsp4 could upregulate pro-inflammatory cytokine and chemokine expression, inhibiting viral replication in vitro. These results provided novel insights for understanding the roles of cytokines in PEDV replication. Topics: Animals; Chemokine CCL2; Chemokine CCL5; Chemokines; Chlorocebus aethiops; Coronavirus Infections; Cytokines; Host-Pathogen Interactions; Interleukin-1beta; Interleukin-8; NF-kappa B; Porcine epidemic diarrhea virus; Swine; Swine Diseases; Tumor Necrosis Factor-alpha; Vero Cells; Viral Nonstructural Proteins; Virus Replication | 2019 |
Astragalus polysaccharides inhibit avian infectious bronchitis virus infection by regulating viral replication.
The avian coronavirus causes infectious bronchitis (IB), which is one of the most serious diseases affecting the avian industry worldwide. However, there are no effective strategies for controlling the IB virus (IBV) at present. Therefore, development of novel antiviral treatment strategies is urgently required. As reported, astragalus polysaccharides (APS) have potential antiviral effects against several viruses; however, the antiviral effect of APS against IBV remains unclear. In this study, we explored whether APS had the potential to inhibit IBV infectionby utilizing several in vitro experimental approaches. To this end, the effect of APS on the replication of IBV was examined in chicken embryo kidney (CEK) cells. Viral titers were calculated by using the plaque formation assay, and the cytotoxicity of APS was tested by utilizing a Cell Counting Kit-8 assay. The expression of viral mRNA and cytokine (IL-1β, IL-6, IL-8 and TNF-α) mRNA transcripts was determined by real-time quantitative RT-PCR(qRT-PCR). IBV titers in infected CEK cells treated with APS were significantly reduced in a dose-dependent manner, indicating that APS inhibited IBV replication in vitro. We also found that the decreased viral replication after APS treatment was associated with reduced mRNA levels of the cytokines IL-1B, IL-6, IL-8 and TNF-α. In conclusion, these results suggest that APS exhibit antiviral activities against IBV and it may represent a potential therapeutic agent for inhibiting the replication of IBV. Topics: Animals; Antiviral Agents; Astragalus Plant; Cell Line; Cell Proliferation; Chickens; Coronavirus Infections; Cytokines; Infectious bronchitis virus; Interleukin-1beta; Interleukin-6; Interleukin-8; Plant Extracts; Polysaccharides; Poultry Diseases; RNA, Messenger; Tumor Necrosis Factor-alpha; Viral Load; Viral Plaque Assay; Virus Replication | 2018 |
Astragalus polysaccharides enhance the immune response to avian infectious bronchitis virus vaccination in chickens.
Astragalus polysaccharides (APS) are biological macromolecules extracted from Astragalus species that have strong immunoregulatory properties. In this study, APS were employed as an adjuvant for an avian infectious bronchitis virus (IBV) vaccine, and its effects on the cellular immune and humoral immune responses to vaccination in chicken were investigated. One hundred and fifty chicken were randomly divided into five groups (n = 30, each group). The chickens in all groups, except for the unvaccinated control group, were vaccinated with an IBV DNA vaccine. Three of the four vaccinated groups were administered different doses of APS (APSL, 10 mg/kg; APSM, 50 mg/kg; and APSH, 100 mg/kg) after the first vaccination, and the remaining vaccinated group served as a control, without any additional treatment. At 14, 28, and 42 days after the first vaccination, serum anti-IBV antibody titers; peripheral lymphocyte proliferation; and the mRNA expression of IL-1β, IL-2, IL-8, and TNF-α in the spleen were assessed by enzyme-linked immunosorbent assay (ELISA), the cell counting kit-8 (CCK-8), and real time quantitative RT-PCR (qRT-PCR), respectively. At most time points, the titer of IBV-specific antibodies, lymphocyte proliferation, and IL-1β, IL-2, IL-8, and TNF-α mRNA expression levels were higher in three APS groups than in the vaccine control group, and these increases were dose-dependent. These data suggest that APS could be used as an adjuvant for IBV vaccination to provide better protection against IBV infection. Topics: Adaptive Immunity; Adjuvants, Immunologic; Animals; Antibodies, Viral; Astragalus Plant; Cell Proliferation; Chickens; Coronavirus Infections; Cytokines; Enzyme-Linked Immunosorbent Assay; Infectious bronchitis virus; Interleukin-1beta; Interleukin-2; Interleukin-8; Lymphocytes; Peptide Fragments; Plant Extracts; Polysaccharides; Poultry Diseases; RNA, Messenger; Spleen; Time Factors; Tumor Necrosis Factor-alpha; Vaccination; Vaccines, DNA; Viral Vaccines | 2017 |
Coronavirus Infections in the Central Nervous System and Respiratory Tract Show Distinct Features in Hospitalized Children.
Coronavirus (CoV) infections induce respiratory tract illnesses and central nervous system (CNS) diseases. We aimed to explore the cytokine expression profiles in hospitalized children with CoV-CNS and CoV-respiratory tract infections.. A total of 183 and 236 hospitalized children with acute encephalitis-like syndrome and respiratory tract infection, respectively, were screened for anti-CoV IgM antibodies. The expression profiles of multiple cytokines were determined in CoV-positive patients.. Anti-CoV IgM antibodies were detected in 22/183 (12.02%) and 26/236 (11.02%) patients with acute encephalitis-like syndrome and respiratory tract infection, respectively. Cytokine analysis revealed that the level of serum granulocyte colony-stimulating factor (G-CSF) was significantly higher in both CoV-CNS and CoV-respiratory tract infection compared with healthy controls. Additionally, the serum level of granulocyte macrophage colony-stimulating factor (GM-CSF) was significantly higher in CoV-CNS infection than in CoV-respiratory tract infection. In patients with CoV-CNS infection, the levels of IL-6, IL-8, MCP-1, and GM-CSF were significantly higher in their cerebrospinal fluid samples than in matched serum samples.. To the best of our knowledge, this is the first report showing a high incidence of CoV infection in hospitalized children, especially with CNS illness. The characteristic cytokine expression profiles in CoV infection indicate the importance of host immune response in disease progression. Topics: Adolescent; Central Nervous System Viral Diseases; Chemokine CCL2; Child; Child, Hospitalized; Child, Preschool; China; Coronavirus; Coronavirus Infections; Cytokines; Disease Progression; Female; Granulocyte Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor; Humans; Immunoglobulin M; Infant; Interleukin-6; Interleukin-8; Male; Respiratory Tract Infections | 2016 |
The effects of Nigella sativa (Ns), Anthemis hyalina (Ah) and Citrus sinensis (Cs) extracts on the replication of coronavirus and the expression of TRP genes family.
Extracts of Anthemis hyalina (Ah), Nigella sativa (Ns) and peels of Citrus sinensis (Cs) have been used as folk medicine to fight antimicrobial diseases. To evaluate the effect of extracts of Ah, Ns and Cs on the replication of coronavirus (CoV) and on the expression of TRP genes during coronavirus infection, HeLa-CEACAM1a (HeLa-epithelial carcinoembryonic antigen-related cell adhesion molecule 1a) cells were inoculated with MHV-A59 (mouse hepatitis virus-A59) at moi of 30. 1/50 dilution of the extracts was found to be the safe active dose. ELISA kits were used to detect the human IL-8 levels. Total RNA was isolated from the infected cells and cDNA was synthesized. Fluidigm Dynamic Array nanofluidic chip 96.96 was used to analyze the mRNA expression of 21 TRP genes and two control genes. Data was analyzed using the BioMark digital array software. Determinations of relative gene expression values were carried out by using the 2(-∆∆Ct) method (normalized threshold cycle (Ct) value of sample minus normalized Ct value of control). TCID50/ml (tissue culture infectious dose that will produce cytopathic effect in 50% of the inoculated tissue culture cells) was found for treatments to determine the viral loads. The inflammatory cytokine IL-8 level was found to increase for both 24 and 48 h time points following Ns extract treatment. TRPA1, TRPC4, TRPM6, TRPM7, TRPM8 and TRPV4 were the genes which expression levels changed significantly after Ah, Ns or Cs extract treatments. The virus load decreased when any of the Ah, Ns or Cs extracts was added to the CoV infected cells with Ah extract treatment leading to undetectable virus load for both 6 and 8 hpi. Although all the extract treatments had an effect on IL-8 secretion, TRP gene expression and virus load after CoV infection, it was the Ah extract treatment that showed the biggest difference in virus load. Therefore Ah extract is the best candidate in our hands that contains potential treatment molecule(s). Topics: Animals; Anthemis; Citrus sinensis; Coronavirus; Coronavirus Infections; Gene Expression Regulation, Viral; HeLa Cells; Humans; Interleukin-8; Medicine, Traditional; Mice; Nigella sativa; Plant Extracts; Transient Receptor Potential Channels; Virus Replication | 2014 |
Quantification of mRNA encoding cytokines and chemokines and assessment of ciliary function in canine tracheal epithelium during infection with canine respiratory coronavirus (CRCoV).
One of the first lines of defence against viral infection is the innate immune response and the induction of antiviral type I interferons (IFNs). However some viruses, including the group 2 coronaviruses, have evolved mechanisms to overcome or circumvent the host antiviral response. Canine respiratory coronavirus (CRCoV) has previously been shown to have a widespread international presence and has been implicated in outbreaks of canine infectious respiratory disease (CIRD). This study aimed to quantify pro-inflammatory cytokine mRNAs following infection of canine air-interface tracheal cultures with CRCoV. Within this system, immunohistochemistry identified ciliated epithelial and goblet cells as positive for CRCoV, identical to naturally infected cases, thus the data obtained would be fully transferable to the situation in vivo. An assay of ciliary function was used to assess potential effects of CRCoV on the mucociliary system. CRCoV was shown to reduce the mRNA levels of the pro-inflammatory cytokines TNF-alpha and IL-6 and the chemokine IL-8 during the 72 h post-inoculation. The mechanism for this is unknown, however the suppression of a key antiviral strategy during a period of physiologic and immunological stress, such as on entry to a kennel, could potentially predispose a dog to further pathogenic challenge and the development of respiratory disease. Topics: Animals; Base Sequence; Chemokines; Cilia; Coronavirus Infections; Coronavirus Nucleocapsid Proteins; Coronavirus, Canine; Cytokines; DNA Primers; Dog Diseases; Dogs; Epithelium; Genes, Viral; Interleukin-6; Interleukin-8; Nucleocapsid Proteins; RNA, Messenger; RNA, Viral; Tissue Culture Techniques; Trachea; Tumor Necrosis Factor-alpha | 2009 |