Respiratory syncytial virus (RSV) entry is inhibited by serine protease inhibitor AEBSF when present during an early stage of infection

Gucht, Winke Van der; Leemans, Annelies; Schryver, Marjorie De; Heykers, Annick; Caljon, Guy; Maes, Louis; Cos, Paul; Delputte, Peter

doi:10.1186/s12985-017-0824-3

Cited by 14 publications

(10 citation statements)

References 48 publications

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“…These findings agree with prior research indicating that components of human milk and digesta, such as proteases, protease inhibitors, and immunoglobulins (SIgA, IgG and IgM), β-casein, lactoferrin and lactoperoxidase, milk fat, cells and bacteria, are capable of inhibiting RSV infectivity ( 4 – 7 , 12 ). Many active proteases, including plasmin, elastase, kallikrein and carboxypeptidases are present in human milk ( 13 ).…”

Section: Resultssupporting

confidence: 92%

“…Human milk contains a broad array of protease inhibitors, including α1-antitrypsin, α1-antichymotrypsin, α2-antiplasmin, plasma serine protease inhibitor and antithrombin III ( 16 ). Serine protease inhibitors [4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF) and N-p-Tosyl-L-phenylalanine chloromethyl ketone (TPCK)] inhibited RSV A2 infection of HEp-2 cells ( 12 ). Host cell proteases are necessary for initiating the infection cycle of RSV through specific cleavage of the F protein into F1 and F2 subunits, which is required for the activation of membrane fusion ( 12 , 17 ).…”

Section: Resultsmentioning

confidence: 99%

“…Serine protease inhibitors [4-(2-Aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF) and N-p-Tosyl-L-phenylalanine chloromethyl ketone (TPCK)] inhibited RSV A2 infection of HEp-2 cells ( 12 ). Host cell proteases are necessary for initiating the infection cycle of RSV through specific cleavage of the F protein into F1 and F2 subunits, which is required for the activation of membrane fusion ( 12 , 17 ). Thus, the presence of protease inhibitors in milk and milk digestive samples could inhibit RSV activity.…”

Section: Resultsmentioning

confidence: 99%

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Purification of Antibodies From Human Milk and Infant Digestates for Viral Inhibition Assays

et al. 2020

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Oral administration of enteric pathogen-specific immunoglobulins may be an ideal approach for preventing infectious diarrhea in infants and children. For oral administration to be effective, antibodies must survive functionally intact within the highly proteolytic digestive tract. As an initial step toward assessing the viability of this approach, we examined the survival of palivizumab, a recombinant monoclonal antibody (IgG1κ), across infant digestion and its ability to neutralize respiratory syncytial virus (RSV). Human milk and infant digestive samples contain substances known to interfere with the RSV neutralization assay (our selected functional test for antibody survival through digestion), therefore, antibody extraction from the matrix was required prior to performing the assay. The efficacy of various approaches for palivizumab purification from human milk, infant's gastric and intestinal digestates, including casein precipitation, salting out, molecular weight cut-off, and affinity chromatography (protein A and G) were compared. Affinity chromatography using protein G with high-salt elution followed by 30-kDa molecular weight cut-off centrifugal filtration was the most effective technique for purifying palivizumab from human milk and infant digestates with a high yield and reduced background interference for the viral neutralization assay. This work is broadly applicable to the optimal isolation of antibodies from human milk and infant digesta for viral neutralization assays, enables the examination of how digestion affects the viral neutralization capacity of antibodies within milk and digestive samples, and paves the way for assessment of the viability of oral administration of recombinant antibodies as a therapeutic approach to prevent enteric pathogen-induced infectious diarrhea in infants.

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Section: Resultssupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Purification of Antibodies From Human Milk and Infant Digestates for Viral Inhibition Assays

et al. 2020

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“…All cells were cultured in Dulbecco’s modified Eagle medium containing 10% inactivated fetal bovine serum (DMEM-10) (Thermo Fisher Scientific, Waltham, MA USA). RSV reference strains A2 and B1 were obtained from BEI resources, RSV A2 was cultivated in HEp-2 cells as described by Van der Gucht W. et al [32] and RSV B1 was cultivated on Vero cells in medium containing 2% inactivated foetal bovine serum (iFBS) until cytopathic effects (CPE) were visible throughout the flask. Virus was collected as described for RSV A2 and quantified in a conventional plaque assay on HEp-2 cells as described by Schepens et al [33].…”

Section: Methodsmentioning

confidence: 99%

Isolation and Characterization of Clinical RSV Isolates in Belgium during the Winters of 2016–2018

Gucht

Stobbelaar

Govaerts

et al. 2019

Viruses

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Respiratory Syncytial Virus (RSV) is a very important viral pathogen in children, immunocompromised and cardiopulmonary diseased patients and the elderly. Most of the published research with RSV was performed on RSV Long and RSV A2, isolated in 1956 and 1961, yet recent RSV isolates differ from these prototype strains. Additionally, these viruses have been serially passaged in cell culture, which may result in adaptations that affect virus–host interactions. We have isolated RSV from mucosal secretions of 12 patients in the winters 2016–2017 and 2017–2018, of which eight RSV-A subtypes and four RSV-B subtypes. Passage 3 of the isolates was assessed for viral replication kinetics and infectious virus production in HEp-2, A549 and BEAS-2B cells, thermal stability at 37 °C, 32 °C and 4 °C, syncytia formation, neutralization by palivizumab and mucin mRNA expression in infected A549 cells. We observed that viruses isolated in one RSV season show differences on the tested assays. Furthermore, comparison with RSV A2 and RSV B1 reveals for some RSV isolates differences in viral replication kinetics, thermal stability and fusion capacity. Major differences are, however, not observed and differences between the recent isolates and reference strains is, overall, similar to the observed variation in between the recent isolates. One clinical isolate (BE/ANT-A11/17) replicated very efficiently in all cell lines, and remarkably, even better than RSV A2 in the HEp-2 cell line.

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“… 1 , 2 It causes lower respiratory tract infection affecting mostly elder people and children. 3 , 4 The RSV virus has 2 main subtypes; subtype A and B. 5 Luminal columnar cells, especially ciliated cells, can be affected by RSV, and the virus can be shed solely from the apical surface of ciliated cells and spread to neighbouring cells by the cilial beats.…”

Section: Introductionmentioning

confidence: 99%

Immunity Cell Responses to RSV and the Role of Antiviral Inhibitors: A Systematic Review

Churiso¹,

Husen²,

Bulbula³

et al. 2022

IDR

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Antigen-presenting cells recognize respiratory syncytial virus antigens, and produce cytokines and chemokines that act on immune cells. Dendritic cells play the main role in inflammatory cytokine responses. Similarly, alveolar macrophages produce IFN-β, IFN-α, TNF-α, IL-6, CXCL10, and CCL3, while alternatively activated macrophages differentiate at the late phase, and require IL-13 or IL-4 cytokines. Furthermore, activated NKT cells secrete IL-13 and IL-4 that cause lung epithelial, endothelial and fibroblasts to secrete eotaxin that enhances the recruitment of eosinophil to the lung. CD8 + and CD4 + T cells infection by the virus decreases the IFN-γ and IL-2 production. Despite this, both are involved in terminating virus replication. CD8 + T cells produce a larger amount of IFN-γ than CD4 + T cells, and CD8 + T cells activated under type 2 conditions produce IL-4, down regulating CD8 expression, granzyme and IFN-γ production. Antiviral inhibitors inhibit biological functions of viral proteins. Some of them directly target the virus replication machinery and are effective at later stages of infection; while others inhibit F protein dependent fusion and syncytium formation. TMC353121 reduces inflammatory cytokines, TNF-α, IL-6, and IL-1β and chemokines, KC, IP-10, MCP and MIP1-α. EDP-938 inhibits viral nucleoprotein (N), while GRP-156784 blocks the activity of respiratory syncytial virus ribonucleic acid (RNA) polymerase. PC786 inhibits non-structural protein 1 (NS-1) gene, RANTES transcripts, virus-induced CCL5, IL-6, and mucin increase. In general, it is an immune reaction that is blamed for the disease severity and pathogenesis in respiratory syncytial virus infection. Anti-viral inhibitors not only inhibit viral entry and replication, but also may reduce inflammatory cytokines and chemokines. Many respiratory syncytial virus inhibitors are proposed; however, only palivizumab and ribavirin are approved for prophylaxis and treatment, respectively. Hence, this review is focused on immunity cell responses to respiratory syncytial virus and the role of antiviral inhibitors.

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Respiratory syncytial virus (RSV) entry is inhibited by serine protease inhibitor AEBSF when present during an early stage of infection

Cited by 14 publications

References 48 publications

Purification of Antibodies From Human Milk and Infant Digestates for Viral Inhibition Assays

Purification of Antibodies From Human Milk and Infant Digestates for Viral Inhibition Assays

Isolation and Characterization of Clinical RSV Isolates in Belgium during the Winters of 2016–2018

Immunity Cell Responses to RSV and the Role of Antiviral Inhibitors: A Systematic Review

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