Pathogenesis Of Severe Acute Respiratory Syndrome Research Articles

The Coronavirus SARS-Cov-2: the Characteristics of Structural Proteins, Contagiousness, and Possible Immune Collisions

Relevance. Coronavirus SARS-Cov-2 is a novel virus demonstrating the ability to be trans¬mitted from human-to-human, via respiratory droplets or close contact, and cause the severe acute respiratory syndrome (SARS). The role of its structural proteins in the SARS pathogenesis is unknown.Aim is to characterize the features of the SARS-Cov-2 structural proteins and their changes associated with acquiring other way of transmission and analyze the possibility of heterologous immunity emergence in its infection. Materials and method. For the computer analysis and alignment, the gene sequences of SARS-Cov-2 , SARS-CoV , MERS-CoV и bat CoV HKU3 reference strains were used from the Internet. From the primary structure of their genes it were translated their structural proteins: spike (S), envelope (E),membrane (M), and nucleocapsid (N). The genetic code of structural proteins was also defined. The search of homologous sequences in the SARS-Cov-2 S-protein, surface proteins of other viruses, and human proteins was made to find immune epitope continuum of protein relationships.Results. In the SARS-Cov-2 structural proteins amino acid sequences of M, E, and N-proteins are conservative. The S1 subunit of the S-protein contains some large insertions, significant changes of the amino acid content with the predominance of arginine and lysine which is typical for the surface glycoproteins in the viruses possessing high contagiousness. The S2 subunit is rather conservative and retain negative polarity. The S-protein exhibits the immune epitope relationships with many proteins of viruses and human which may be associated with immune collisions.Conclusion: The SARSCov-2 features are determined by marked changes of the S1 subunit structure in the S-protein which may be responsible for its contagiousness and many immune collisions aggravating infection process.

The Coronavirus SARS-Cov-2: the Characteristics of Structural Proteins, Contagiousness, and Possible Immune Collisions

Relevance. Coronavirus SARS-Cov-2 is a novel virus demonstrating the ability to be trans¬mitted from human-to-human, via respiratory droplets or close contact, and cause the severe acute respiratory syndrome (SARS). The role of its structural proteins in the SARS pathogenesis is unknown.Aim is to characterize the features of the SARS-Cov-2 structural proteins and their changes associated with acquiring other way of transmission and analyze the possibility of heterologous immunity emergence in its infection. Materials and method. For the computer analysis and alignment, the gene sequences of SARS-Cov-2 , SARS-CoV , MERS-CoV и bat CoV HKU3 reference strains were used from the Internet. From the primary structure of their genes it were translated their structural proteins: spike (S), envelope (E),membrane (M), and nucleocapsid (N). The genetic code of structural proteins was also defined. The search of homologous sequences in the SARS-Cov-2 S-protein, surface proteins of other viruses, and human proteins was made to find immune epitope continuum of protein relationships.Results. In the SARS-Cov-2 structural proteins amino acid sequences of M, E, and N-proteins are conservative. The S1 subunit of the S-protein contains some large insertions, significant changes of the amino acid content with the predominance of arginine and lysine which is typical for the surface glycoproteins in the viruses possessing high contagiousness. The S2 subunit is rather conservative and retain negative polarity. The S-protein exhibits the immune epitope relationships with many proteins of viruses and human which may be associated with immune collisions.Conclusion: The SARSCov-2 features are determined by marked changes of the S1 subunit structure in the S-protein which may be responsible for its contagiousness and many immune collisions aggravating infection process.

Science in the fight against the novel coronavirus disease 2019 (COVID-19).

Science in the fight against the novel coronavirus disease 2019 (COVID-19).

SARS-CoV Regulates Immune Function-Related Gene Expressions in Human Monocytic Cells

Abstract Background: Severe Acute Respiratory Syndrome (SARS) is characterized by acute respiratory distress (ARDS) and pulmonary fibrosis, and the monocyte/macrophage is the key player in the pathogenesis of SARS. Methods: In this study, we compared the transcriptional profiles of SARS coronavirus (SARS-CoV) infected monocytic cells against that infected by coronavirus 229E (CoV-229E). Total RNA was extracted from infected DC-SIGN transfected monocytes (THP-1-DC-SIGN) at 6 and 24 h after infection and the gene expression was profiled by oligonucleotide-based microarray. Results: Analysis of immune-related gene expression profiles showed that 24 h after SARS-CoV infection, (i) IFN-alpha/beta-inducible and cathepsin/proteosome genes were down-regulated; (ii) the hypoxia/hyperoxia-related genes were up-regulated; and (iii) the TLR/TLR-signaling, cytokine/cytokine receptor-related, chemokine/chemokine receptor-related, the lysosome-related, MHC/chaperon-related, and fibrosis-related genes were differentially regulated. Conclusion: These results elucidate that monocyte/macrophage dysfunction and dysregulation of fibrosis-related genes are two important pathogenic events of SARS.

Differential characteristics of the early stage of lung inflammation induced by SARS-CoV Nucleocapsid protein related to age in the mouse

.Objective:Severe acute respiratory syndrome (SARS) is an acute infectious disease of the respiratory system which has newly emerged. Interestingly, it appears to be a disease that predominantly affects adults while the mortality in children is extremely low. However, the pathogenesis of SARS in relation to different characteristics relevant to age remains unclear.Material and Methods:To better understand the role of cytokines in the immunopathological processes of SARS, weanling (4 weeks old), young (6 weeks old) and adult (10 weeks old) male BALB/C mice were inoculated intranasally with N-protein of SARS-CoV in this study. Serum or lung homogenate levels of some cytokines such as tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) along with acute injury lung index and histology were also analyzed.Results:Histopathological analysis of adult male BALB/C mice after N-protein infection showed progressive inflammatory reactions, especially pulmonary edema, in accordance with a moderately (~13%) elevated level of W/D ratio at 24 h. Although adult groups underwent a progressive lung inflammation in the acute phase accompanied by raised levels of TNF-α in serum, no significant changes in lung TNF-α level were reported simultaneously. Moreover, adult SARS infected BALB/c mice showed elevated levels of IFN-γ while IFN-γ levels in weanling and young groups had no obvious association with lung inflammation.Conclusion:Our study supports the observation that adult mice do have progressively greater immune reactions than weanling and adolescent ones over time. The relative immaturity of the immune system in weanlings may confer benefit leading to less impairment of lung function. However, the measurement of TNF-α and IFN-γ levels was not indicative of the severity of lung injury at the early stage of disease.

Mechanisms of severe acute respiratory syndrome pathogenesis and innate immunomodulation.

The modulation of the immune response is a common practice of many highly pathogenic viruses. The emergence of the highly pathogenic coronavirus severe acute respiratory virus (SARS-CoV) serves as a robust model system to elucidate the virus-host interactions that mediate severe end-stage lung disease in humans and animals. Coronaviruses encode the largest positive-sense RNA genome of approximately 30 kb, encode a variety of replicase and accessory open reading frames that are structurally unique, and encode novel enzymatic functions among RNA viruses. These viruses have broad or specific host ranges, suggesting the possibility of novel strategies for targeting and regulating host innate immune responses following virus infection. Using SARS-CoV as a model, we review the current literature on the ability of coronaviruses to interact with and modify the host intracellular environment during infection. These studies are revealing a rich set of novel viral proteins that engage, modify, and/or disrupt host cell signaling and nuclear import machinery for the benefit of virus replication.

Human LINE1 endonuclease domain as a putative target of SARS-associated autoantibodies involved in the pathogenesis of severe acute respiratory syndrome

Severe acute respiratory syndrome (SARS) is a disease with a mortality of 9.56%. Although SARS is etiologically linked to a new coronavirus (SARS-CoV) and functional cell receptor has been identified, the pathogenesis of the virus infection is largely unclear. The clinical specimens were processed and analyzed using an indirect enzyme-linked immunosorbent assay (ELISA) in-house. Further investigations of target antigen included reviews of phage display technique, rapid amplification of cDNA ends (RACE) technique, protein expression and purification, Western blotting validation, serological and immunohistochemical staining in postmortem tissue. A type of medium or low titer anti-lung tissue antibodies were found in the sera of SARS patients at the early stage of the disease. Human long interspersed nuclear element 1 (LINE1) gene endonuclease (EN) domain protein was one of the target autoantigens and it was aberrantly expressed in the lung tissue of SARS patients. Anti-EN antibody was positive in the sera of 40.9% of SARS patients. Human LINE1 endonuclease domain was identified as a putative target of SARS-associated autoantibodies, which were presented in the serum of SARS patients and may be involved in the pathogenesis of SARS.

Pathological Changes in Masked Palm Civets Experimentally Infected by Severe Acute Respiratory Syndrome (SARS) Coronavirus

SummaryMasked palm civets are highly susceptible to infection with the severe acute respiratory syndrome coronavirus (SARS-CoV). Infected animals become less aggressive and develop pyrexia, lethargy and diarrhoea. The present study describes the spectrum of histopathological changes in the lung, spleen, lymph node, liver, small intestine, kidney and cerebrum of civets infected experimentally with SARS-CoV. In-situ hybridization (ISH) with probes specific for the RNA polymerase gene demonstrated viral RNA in the lung, small intestine and cerebrum only. In-situ labelling was employed in order to demonstrate cellular apoptosis in the cerebrum, but there was no evidence of apoptosis within the myocardium. These results indicate that SARS-CoV causes multi-organ pathology in civets, similar to that observed in human SARS patients. These parallels suggest that civets may be used as an animal model of this infection to gain insight into the pathogenesis of SARS and for evaluation of candidate vaccines and antiviral drugs.

Mechanisms of Severe Acute Respiratory Syndrome Pathogenesis and Innate Immunomodulation

Mechanisms of Severe Acute Respiratory Syndrome Pathogenesis and Innate Immunomodulation

The association of RANTES polymorphism with severe acute respiratory syndrome in Hong Kong and Beijing Chinese

BackgroundChemokines play important roles in inflammation and antiviral action. We examined whether polymorphisms of RANTES, IP-10 and Mig affect the susceptibility to and outcome of severe acute respiratory syndrome (SARS).MethodsWe tested the polymorphisms of RANTES, IP-10 and Mig for their associations with SARS in 495 Hong Kong Chinese SARS patients and 578 controls. Then we tried to confirm the results in 356 Beijing Chinese SARS patients and 367 controls.ResultsRANTES -28 G allele was associated with SARS susceptibility in Hong Kong Chinese (P < 0.0001, OR = 2.80, 95%CI:2.11–3.71). Individuals with RANTES -28 CG and GG genotypes had a 3.28-fold (95%CI:2.32–4.64) and 3.06-fold (95%CI:1.47–6.39) increased risk of developing SARS respectively (P < 0.0001). This -28 G allele conferred risk of death in a gene-dosage dependent manner (P = 0.014) with CG and GG individuals having a 2.12-fold (95% CI: 1.11–4.06) and 4.01-fold (95% CI: 1.30–12.4) increased risk. For the replication of RANTES data in Beijing Chinese, the -28 G allele was not associated with susceptibility to SARS. However, -28 CG (OR = 4.27, 95%CI:1.64–11.1) and GG (OR = 3.34, 95%CI:0.37–30.7) were associated with admission to intensive care units or death due to SARS (P = 0.011).ConclusionRANTES -28 G allele plays a role in the pathogenesis of SARS.

Up-regulation of IL-6 and TNF-α induced by SARS-coronavirus spike protein in murine macrophages via NF-κB pathway

The clinical picture of severe acute respiratory syndrome (SARS) is characterized by an over-exuberant immune response with lung lymphomononuclear cells infilteration and proliferation that may account for tissue damage more than the direct effect of viral replication. To understand how cells response in the early stage of virus–host cell interaction, in this study, a purified recombinant S protein was studied for stimulating murine macrophages (RAW264.7) to produce proinflammatory cytokines (IL-6 and TNF-α) and chemokine IL-8. We found that direct induction of IL-6 and TNF-α release in the supernatant in a dose-, time-dependent manner and highly spike protein-specific, but no induction of IL-8 was detected. Further experiments showed that IL-6 and TNF-α production were dependent on NF-κB, which was activated through I-κBα degradation. These results suggest that SARS-CoV spike protein may play an important role in the pathogenesis of SARS, especially in inflammation and high fever.

Pathology and Pathogenesis of Severe Acute Respiratory Syndrome

Severe acute respiratory syndrome (SARS) is an emerging infectious viral disease characterized by severe clinical manifestations of the lower respiratory tract. The pathogenesis of SARS is highly complex, with multiple factors leading to severe injury in the lungs and dissemination of the virus to several other organs. The SARS coronavirus targets the epithelial cells of the respiratory tract, resulting in diffuse alveolar damage. Several organs/cell types may be infected in the course of the illness, including mucosal cells of the intestines, tubular epithelial cells of the kidneys, neurons of the brain, and several types of immune cells, and certain organs may suffer from indirect injury. Extensive studies have provided a basic understanding of the pathogenesis of this disease. In this review we describe the most significant pathological features of SARS, explore the etiological factors causing these pathological changes, and discuss the major pathogenetic mechanisms. The latter include dysregulation of cytokines/chemokines, deficiencies in the innate immune response, direct infection of immune cells, direct viral cytopathic effects, down-regulation of lung protective angiotensin converting enzyme 2, autoimmunity, and genetic factors. It seems that both abnormal immune responses and injury to immune cells may be key factors in the pathogenesis of this new disease.

Developing MHV-1 infection as an animal model for Severe Acute Respiratory Syndrome (SARS) (43.30)

Abstract SARS is a life threatening infectious disease that acquired epidemic proportions in late 2002 affecting 8000 people worldwide and claiming nearly 800 lives. It is caused by a novel group 2 coronavirus HCoV-SARS. No suitable small animal model currently exists for this disease. Developing a good animal model is crucial for generating effective anti viral therapies. Preliminary data indicate that intranasal infection of A/J mice with a related coronavirus MHV1 closely mimics the disease observed in humans infected with HCoV-SARS. Following infection these mice experience severe weight loss and the lungs show evidence of patchy mononuclear infiltrates and pneumonitis. Significant mortality is observed beginning around day 6–7 post infection suggesting an adaptive immune component to the pathogenesis of the disease. Interestingly, wild type BALB/c and B6 mice are relatively resistant to disease however, resistance is compromised in BALB/c mice lacking both perforin and IFNγ, and B6 mice deficient in receptors to IFNαβ. Our overall goal is to characterize the phenotype and function of the virus-specific T cell response in susceptible strains of mice to ascertain if a dysregulated T cell response contributes to morbidity and mortality. We believe this study will provide useful information regarding the pathogenesis of SARS and aid translational initiatives aimed at infection control and vaccine design

Expression of elevated levels of pro‐inflammatory cytokines in SARS‐CoV‐infected ACE2+ cells in SARS patients: relation to the acute lung injury and pathogenesis of SARS

The authors have previously shown that acute lung injury (ALI) produces a wide spectrum of pathological processes in patients who die of severe acute respiratory syndrome (SARS) and that the SARS coronavirus (SARS‐CoV) nucleoprotein is detectable in the lungs, and other organs and tissues, in these patients. In the present study, immunohistochemistry (IHC) and in situ hybridization (ISH) assays were used to analyse the expression of angiotensin‐converting enzyme 2 (ACE2), SARS‐CoV spike (S) protein, and some pro‐inflammatory cytokines (PICs) including MCP‐1, TGF‐β1, TNF‐α, IL‐1β, and IL‐6 in autopsy tissues from four patients who died of SARS. SARS‐CoV S protein and its RNA were only detected in ACE2+ cells in the lungs and other organs, indicating that ACE2‐expressing cells are the primary targets for SARS‐CoV infection in vivo in humans. High levels of PICs were expressed in the SARS‐CoV‐infected ACE2+ cells, but not in the uninfected cells. These results suggest that cells infected by SARS‐CoV produce elevated levels of PICs which may cause immuno‐mediated damage to the lungs and other organs, resulting in ALI and, subsequently, multi‐organ dysfunction. Therefore application of PIC antagonists may reduce the severity and mortality of SARS. Copyright © 2006 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Inflammation inhibitors were remarkably up-regulated in plasma of severe acute respiratory syndrome patients at progressive phase

Severe acute respiratory syndrome (SARS) is a severe infectious disease that has affected many countries and regions since 2002. A novel member of the coronavirus, SARS‐CoV, has been identified as the causative agent. However, the pathogenesis of SARS is still elusive. In this study, we used 2‐D DIGE and MS to analyze the protein profiles of plasma from SARS patients, in the search for proteomic alterations associated with the disease progression, which could provide some clues to the pathogenesis. To enrich the low‐abundance proteins in human plasma, two highly abundant proteins, albumin and IgG, were first removed. By comparing the plasma proteins of SARS patients with those of a normal control group, several proteins with a significant alteration were found. The up‐regulated proteins were identified as alpha‐1 acid glycoprotein, haptoglobin, alpha‐1 anti‐chymotrypsin and fetuin. The down‐regulated proteins were apolipoprotein A‐I, transferrin and transthyretin. Most of the proteins showed significant changes (up‐ or down‐regulated) in the progressive phase of disease, and there was a trend back to normal level during the convalescent phase. Among these proteins, the alterations of fetuin and anti‐chymotrypsin were further confirmed by Western blotting. Since all the up‐regulated proteins identified above are well‐known inflammation inhibitors, these results strongly suggest that the body starts inflammation inhibition to sustain the inflammatory response balance in the progression of SARS.

How the SARS coronavirus causes disease: host or organism?

The previous epidemic of severe acute respiratory syndrome (SARS) has ended. However, many questions concerning how the aetiological agent, the novel SARS coronavirus (CoV), causes illness in humans remain unanswered. The pathology of fatal cases of SARS is dominated by diffuse alveolar damage. Specific histological changes are not detected in other organs. These contrast remarkably with the clinical picture, in which there are apparent manifestations in multiple organs. Both pathogen and host factors are important in the pathogenesis of SARS. The choice of specific receptors and the unique genome of the SARS‐CoV are important elements in understanding the biology of the pathogen. For the host cells, the outcome of SARS‐CoV infection, whether there are cytopathic effects or not, depends on the cell types that are infected. At the whole‐body level, immune‐mediated damage, due to activation of cytokines and/or chemokines and, perhaps, autoimmunity, may play key roles in the clinical and pathological features of SARS. Continued research is still required to determine the pathogenetic mechanisms involved and to combat this new emerging human infectious disease. Copyright © 2006 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Immunopathogenesis of coronavirus infections: implications for SARS

At the end of 2002, the first cases of severe acute respiratory syndrome (SARS) were reported, and in the following year, SARS resulted in considerable mortality and morbidity worldwide. SARS is caused by a novel species of coronavirus (SARS-CoV) and is the most severe coronavirus-mediated human disease that has been described so far. On the basis of similarities with other coronavirus infections, SARS might, in part, be immune mediated. As discussed in this Review, studies of animals that are infected with other coronaviruses indicate that excessive and sometimes dysregulated responses by macrophages and other pro-inflammatory cells might be particularly important in the pathogenesis of disease that is caused by infection with these viruses. It is hoped that lessons from such studies will help us to understand more about the pathogenesis of SARS in humans and to prevent or control outbreaks of SARS in the future.

Lung pathology and pathogenesis of severe acute respiratory syndrome: a report of six full autopsies

Severe acute respiratory syndrome (SARS) is an emerging infectious disease that first manifested in humans in November 2002. The SARS-associated coronavirus (SARS-CoV) has been identified as the causal agent, but the pathology and pathogenesis are still not quite clear. Post-mortem lung samples from six patients who died from SARS from April to July 2003 were studied by light and electron microscopy, Masson trichromal staining and immunohistochemistry. Evidence of infection with the SARS-CoV was determined by reverse-transcription PCR (RT-PCR) , serological examination and electron microscopy. Four of six patients had serological and RT-PCR evidence of recent infection of SARS-CoV. Morphologic changes are summarized as follows: (1) Diffuse and bilateral lung consolidation was seen in all patients (6/6) with increasing lung weight. (2) Diffuse alveolar damage was universal (6/6) with hyaline membrane formation (6/6), intra-alveolar edema/hemorrhage (6/6), fibrin deposition (6/6), pneumocyte desquamation (6/6). A marked disruption in the integrity of the alveolar epithelium was confirmed by immunostaining for the epithelial marker AE1/AE3 (6/6). (3) Type II pneumocytes, with mild hyperplasia, atypia, cytomegaly with granular amphophilic cytoplasm and intracytoplasmic lipid accumulation (5/6). (4) Giant cells in the alveoli were seen in five of 6 patients (5/6) , most of which were positive for the epithelial marker AE1/AE3 (5/6), but some cells were positive for the macrophage marker CD68(2/6). (5) A pronounced increase of macrophages were seen in the alveoli and the interstitium of the lung (6/6), which was confirmed by histological study and immunohistochemistry. (6) Haemophagocytosis was present in five of the 6 patients(5/6). (7) Lung fibrosis was seen in five patients(5/6), with alveolar septa and interstitium thickening(5/6), intraalveolar organizing exudates (6/6) and pleura thickening (4/6). Proliferation of collagen was confirmed by Masson trichromal staining, most of which was type III collagen by immunostaining. The formation of distinctive fibroblast/myofibroblast foci was seen in five patients (5/6) by light microscopy and immunochemistry. (8) Squamous metaplasia of bronchial mucosa was seen in five patients(5/6). (9) Thrombi was seen in all patients(6/6). (10) Accompanying infection was present in two patients, one was bacteria, the other was fungus. In addition, electron microscopy revealed viral particles in the cytoplasm of alveolar epithelial cells and endothelial cells corresponding to coronavirus. Direct injury of SARS-CoV on alveolar epithelium, prominent macrophage infiltration and distinctive fibroblast/myofibroblast proliferation may play major roles in the pathogenesis of SARS.

Multiple organ infection and the pathogenesis of SARS

After >8,000 infections and >700 deaths worldwide, the pathogenesis of the new infectious disease, severe acute respiratory syndrome (SARS), remains poorly understood. We investigated 18 autopsies of patients who had suspected SARS; 8 cases were confirmed as SARS. We evaluated white blood cells from 22 confirmed SARS patients at various stages of the disease. T lymphocyte counts in 65 confirmed and 35 misdiagnosed SARS cases also were analyzed retrospectively. SARS viral particles and genomic sequence were detected in a large number of circulating lymphocytes, monocytes, and lymphoid tissues, as well as in the epithelial cells of the respiratory tract, the mucosa of the intestine, the epithelium of the renal distal tubules, the neurons of the brain, and macrophages in different organs. SARS virus seemed to be capable of infecting multiple cell types in several organs; immune cells and pulmonary epithelium were identified as the main sites of injury. A comprehensive theory of pathogenesis is proposed for SARS with immune and lung damage as key features.

Antibody to severe acute respiratory syndrome (SARS)-associated coronavirus spike protein domain 2 cross-reacts with lung epithelial cells and causes cytotoxicity

Both viral effect and immune-mediated mechanism are involved in the pathogenesis of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) infection. In this study, we showed that in SARS patient sera there were autoantibodies (autoAbs) that reacted with A549 cells, the type-2 pneumocytes, and that these autoAbs were mainly IgG. The autoAbs were detectable 20 days after fever onset. Tests of non-SARS-pneumonia patients did not show the same autoAb production as in SARS patients. After sera IgG bound to A549 cells, cytotoxicity was induced. Cell cytotoxicity and the anti-epithelial cell IgG level were positively correlated. Preabsorption and binding assays indicated the existence of cross-reactive epitopes on SARS-CoV spike protein domain 2 (S2). Furthermore, treatment of A549 cells with anti-S2 Abs and IFN-γ resulted in an increase in the adherence of human peripheral blood mononuclear cells to these epithelial cells. Taken together, we have demonstrated that the anti-S2 Abs in SARS patient sera cause cytotoxic injury as well as enhance immune cell adhesion to epithelial cells. The onset of autoimmune responses in SARS-CoV infection may be implicated in SARS pathogenesis.