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Article ; Online: Influenza breakthrough infection in vaccinated mice is characterized by non-pathological lung eosinophilia.

Chang, Lauren A / Choi, Angela / Rathnasinghe, Raveen / Warang, Prajakta / Noureddine, Moataz / Jangra, Sonia / Chen, Yong / De Geest, Bruno G / Schotsaert, Michael

Frontiers in immunology

2023 Volume 14, Page(s) 1217181

Abstract: Eosinophils are important mediators of mucosal tissue homeostasis, anti-helminth responses, and allergy. Lung eosinophilia has previously been linked to aberrant Type 2-skewed T cell responses to respiratory viral infection and may also be a consequence ... ...

Abstract	Eosinophils are important mediators of mucosal tissue homeostasis, anti-helminth responses, and allergy. Lung eosinophilia has previously been linked to aberrant Type 2-skewed T cell responses to respiratory viral infection and may also be a consequence of vaccine-associated enhanced respiratory disease (VAERD), particularly in the case of respiratory syncytial virus (RSV) and the formalin-inactivated RSV vaccine. We previously reported a dose-dependent recruitment of eosinophils to the lungs of mice vaccinated with alum-adjuvanted trivalent inactivated influenza vaccine (TIV) following a sublethal, vaccine-matched H1N1 (A/New Caledonia/20/1999; NC99) influenza challenge. Given the differential role of eosinophil subset on immune function, we conducted the investigations herein to phenotype the lung eosinophils observed in our model of influenza breakthrough infection. Here, we demonstrate that eosinophil influx into the lungs of vaccinated mice is adjuvant- and sex-independent, and only present after vaccine-matched sublethal influenza challenge but not in mock-challenged mice. Furthermore, vaccinated and challenged mice had a compositional shift towards more inflammatory eosinophils (iEos) compared to resident eosinophils (rEos), resembling the shift observed in ovalbumin (OVA)-sensitized allergic control mice, however without any evidence of enhanced morbidity or aberrant inflammation in lung cytokine/chemokine signatures. Furthermore, we saw a lung eosinophil influx in the context of a vaccine-mismatched challenge. Additional layers of heterogeneity in the eosinophil compartment were observed via unsupervised clustering analysis of flow cytometry data. Our collective findings are a starting point for more in-depth phenotypic and functional characterization of lung eosinophil subsets in the context of vaccine- and infection-induced immunity.
MeSH term(s)	Animals ; Mice ; Asthma ; Breakthrough Infections ; Hypersensitivity ; Influenza A Virus, H1N1 Subtype ; Influenza Vaccines ; Influenza, Human ; Lung ; Pulmonary Eosinophilia
Chemical Substances	Influenza Vaccines
Language	English
Publishing date	2023-08-04
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2023.1217181
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Novel influenza A viruses in pigs with zoonotic potential, Chile.

Tapia, Rodrigo / Brito, Bárbara / Saavedra, Marco / Mena, Juan / García-Salum, Tamara / Rathnasinghe, Raveen / Barriga, Gonzalo / Tapia, Karla / García, Victoria / Bucarey, Sergio / Jang, Yunho / Wentworth, David / Torremorell, Montserrat / Neira, Víctor / Medina, Rafael A

Microbiology spectrum

2024 Volume 12, Issue 4, Page(s) e0218123

Abstract: Novel H1N2 and H3N2 swine influenza A viruses (IAVs) have recently been identified in Chile. The objective of this study was to evaluate their zoonotic potential. We perform phylogenetic analyses to determine the genetic origin and evolution of these ... ...

Abstract	Novel H1N2 and H3N2 swine influenza A viruses (IAVs) have recently been identified in Chile. The objective of this study was to evaluate their zoonotic potential. We perform phylogenetic analyses to determine the genetic origin and evolution of these viruses, and a serological analysis to determine the level of cross-protective antibodies in the human population. Eight genotypes were identified, all with pandemic H1N1 2009-like internal genes. H1N1 and H1N2 were the subtypes more commonly detected. Swine H1N2 and H3N2 IAVs had hemagglutinin and neuraminidase lineages genetically divergent from IAVs reported worldwide, including human vaccine strains. These genes originated from human seasonal viruses were introduced into the swine population since the mid-1980s. Serological data indicate that the general population is susceptible to the H3N2 virus and that elderly and young children also lack protective antibodies against the H1N2 strains, suggesting that these viruses could be potential zoonotic threats. Continuous IAV surveillance and monitoring of the swine and human populations is strongly recommended.IMPORTANCEIn the global context, where swine serve as crucial intermediate hosts for influenza A viruses (IAVs), this study addresses the pressing concern of the zoonotic potential of novel reassortant strains. Conducted on a large scale in Chile, it presents a comprehensive account of swine influenza A virus diversity, covering 93.8% of the country's industrialized swine farms. The findings reveal eight distinct swine IAV genotypes, all carrying a complete internal gene cassette of pandemic H1N1 2009 origin, emphasizing potential increased replication and transmission fitness. Genetic divergence of H1N2 and H3N2 IAVs from globally reported strains raises alarms, with evidence suggesting introductions from human seasonal viruses since the mid-1980s. A detailed serological analysis underscores the zoonotic threat, indicating susceptibility in the general population to swine H3N2 and a lack of protective antibodies in vulnerable demographics. These data highlight the importance of continuous surveillance, providing crucial insights for global health organizations.
MeSH term(s)	Child ; Humans ; Animals ; Swine ; Child, Preschool ; Aged ; Influenza A virus/genetics ; Influenza A Virus, H3N2 Subtype/genetics ; Orthomyxoviridae Infections/epidemiology ; Orthomyxoviridae Infections/veterinary ; Influenza A Virus, H1N1 Subtype/genetics ; Phylogeny ; Chile/epidemiology ; Reassortant Viruses/genetics ; Swine Diseases/epidemiology ; Influenza, Human/epidemiology
Language	English
Publishing date	2024-03-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	2807133-5
ISSN	2165-0497 ; 2165-0497
ISSN (online)	2165-0497
ISSN	2165-0497
DOI	10.1128/spectrum.02181-23
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Shed the light on virus: virucidal effects of 405 nm visible light on SARS-CoV-2 and influenza A virus.

Yahnke, Clifford J / Rathnasinghe, Raveen J / Garcia-Sastre, Adolfo

bioRxiv

Abstract: Germicidal potential of specific wavelengths within the electromagnetic spectrum is an area of growing interest. While ultra-violet (UV) based technologies have shown satisfactory virucidal potential, the photo-toxicity in humans coupled with UV ... ...

Abstract	Germicidal potential of specific wavelengths within the electromagnetic spectrum is an area of growing interest. While ultra-violet (UV) based technologies have shown satisfactory virucidal potential, the photo-toxicity in humans coupled with UV associated polymer degradation limit its use in occupied spaces. Alternatively, longer wavelengths with less irradiation energy such as visible light (405 nm) have largely been explored in the context of bactericidal and fungicidal applications. Such studies indicated that 405 nm mediated inactivation is caused by the absorbance of porphyrins within the organism creating reactive oxygen species which result in free radical damage to its DNA and disruption of cellular functions. The virucidal potential of visible-light based technologies has been largely unexplored and speculated to be not effective given the lack of porphyrins in viruses. The current study demonstrated increased susceptibility of lipid-enveloped respiratory pathogens of importance such as SARS-CoV-2 (causative agent of COVID-19) as well as the influenza A virus to 405nm, visible light in the absence of exogenous photosensitizers, indicating a potential porphyrin-independent alternative mechanism of visible light mediated viral inactivation. Given that visible light is generally safe to humans, our results support further exploration of the use of visible light technology for the application of continuous decontamination in areas within hospitals and/or infectious disease laboratories, specifically for the inactivation of respiratory pathogens such as SARS-CoV-2 and Influenza A.
Keywords	covid19
Language	English
Publishing date	2021-03-15
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2021.03.14.435337
Database	COVID19

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Article ; Online: The virucidal effects of 405 nm visible light on SARS-CoV-2 and influenza A virus.

Rathnasinghe, Raveen / Jangra, Sonia / Miorin, Lisa / Schotsaert, Michael / Yahnke, Clifford / Garcίa-Sastre, Adolfo

Scientific reports

2021 Volume 11, Issue 1, Page(s) 19470

Abstract: The germicidal potential of specific wavelengths within the electromagnetic spectrum is an area of growing interest. While ultra-violet (UV) based technologies have shown satisfactory virucidal potential, the photo-toxicity in humans coupled with UV ... ...

Abstract	The germicidal potential of specific wavelengths within the electromagnetic spectrum is an area of growing interest. While ultra-violet (UV) based technologies have shown satisfactory virucidal potential, the photo-toxicity in humans coupled with UV associated polymer degradation limit their use in occupied spaces. Alternatively, longer wavelengths with less irradiation energy such as visible light (405 nm) have largely been explored in the context of bactericidal and fungicidal applications. Such studies indicated that 405 nm mediated inactivation is caused by the absorbance of porphyrins within the organism creating reactive oxygen species which result in free radical damage to its DNA and disruption of cellular functions. The virucidal potential of visible-light based technologies has been largely unexplored and speculated to be ineffective given the lack of porphyrins in viruses. The current study demonstrated increased susceptibility of lipid-enveloped respiratory pathogens of importance such as SARS-CoV-2 (causative agent of COVID-19) and influenza A virus to 405 nm, visible light in the absence of exogenous photosensitizers thereby indicating a potential alternative porphyrin-independent mechanism of visible light mediated viral inactivation. These results were obtained using less than expected irradiance levels which are considered safe for humans and commercially achievable. Our results support further exploration of the use of visible light technology for the application of continuous decontamination in occupied areas within hospitals and/or infectious disease laboratories, specifically for the inactivation of respiratory pathogens such as SARS-CoV-2 and Influenza A.
MeSH term(s)	Disinfection/instrumentation ; Disinfection/methods ; Dose-Response Relationship, Radiation ; Encephalomyocarditis virus/radiation effects ; Influenza A Virus, H1N1 Subtype/radiation effects ; Light ; SARS-CoV-2/radiation effects ; Time Factors ; Virus Inactivation/radiation effects
Language	English
Publishing date	2021-09-30
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-97797-0
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Replication-Competent ΔNS1 Influenza A Viruses Expressing Reporter Genes

Nogales, Aitor / Schotsaert, Michael / Rathnasinghe, Raveen / DeDiego, Marta L / García-Sastre, Adolfo / Martinez-Sobrido, Luis

Viruses. 2021 Apr. 17, v. 13, no. 4

2021

Abstract: The influenza A virus (IAV) is able to infect multiple mammalian and avian species, and in humans IAV is responsible for annual seasonal epidemics and occasional pandemics of respiratory disease with significant health and economic impacts. Studying IAV ... ...

Abstract	The influenza A virus (IAV) is able to infect multiple mammalian and avian species, and in humans IAV is responsible for annual seasonal epidemics and occasional pandemics of respiratory disease with significant health and economic impacts. Studying IAV involves laborious secondary methodologies to identify infected cells. Therefore, to circumvent this requirement, in recent years, multiple replication-competent infectious IAV expressing traceable reporter genes have been developed. These IAVs have been very useful for in vitro and/or in vivo studies of viral replication, identification of neutralizing antibodies or antivirals, and in studies to evaluate vaccine efficacy, among others. In this report, we describe, for the first time, the generation and characterization of two replication-competent influenza A/Puerto Rico/8/1934 H1N1 (PR8) viruses where the viral non-structural protein 1 (NS1) was substituted by the monomeric (m)Cherry fluorescent or the NanoLuc luciferase (Nluc) proteins. The ΔNS1 mCherry was able to replicate in cultured cells and in Signal Transducer and Activator of Transcription 1 (STAT1) deficient mice, although at a lower extent than a wild-type (WT) PR8 virus expressing the same mCherry fluorescent protein (WT mCherry). Notably, expression of either reporter gene (mCherry or Nluc) was detected in infected cells by fluorescent microscopy or luciferase plate readers, respectively. ΔNS1 IAV expressing reporter genes provide a novel approach to better understand the biology and pathogenesis of IAV, and represent an excellent tool to develop new therapeutic approaches against IAV infections.
Keywords	Influenza A virus ; antiviral agents ; birds ; fluorescence ; fluorescence microscopy ; fluorescent proteins ; influenza ; luciferase ; pathogenesis ; reporter genes ; signal transduction ; therapeutics ; transactivators ; vaccines ; viral nonstructural proteins ; virus replication ; viruses
Language	English
Dates of publication	2021-0417
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	2516098-9
ISSN	1999-4915
ISSN	1999-4915
DOI	10.3390/v13040698
Database	NAL-Catalogue (AGRICOLA)

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Article: A multicomponent intranasal adjuvant drives durable humoral, cellular, and mucosal immune responses to SARS-CoV-2 in young and aged mice.

Jangra, Sonia / Landers, Jeffrey J / Laghlali, Gabriel / Rathnasinghe, Raveen / O'Konek, Jessica J / Janczak, Katarzyna W / García-Sastre, Adolfo / Baker, James R / Schotsaert, Michael / Wong, Pamela T

Research square

2023

Abstract: Multiple FDA-approved SARS-CoV-2 vaccines provide excellent protection against severe disease. Despite this, immunity can wane relatively fast, particularly in the elderly and novel viral variants capable of evading infection- and vaccination-induced ... ...

Abstract	Multiple FDA-approved SARS-CoV-2 vaccines provide excellent protection against severe disease. Despite this, immunity can wane relatively fast, particularly in the elderly and novel viral variants capable of evading infection- and vaccination-induced immunity continue to emerge. Intranasal (IN) vaccination more effectively induces mucosal immune responses than parenteral vaccines, which would improve protection and reduce viral transmission. Here, we developed a rationally designed IN adjuvant consisting of a combined nanoemulsion (NE)-based adjuvant and an RNA-based RIG-I agonist (IVT DI) to drive more robust, broadly protective antibody and T cell responses. We previously demonstrated this combination adjuvant (NE/IVT) potently induces protective immunity through synergistic activation of an array of innate receptors. We now demonstrate that NE/IVT with the SARS-CoV-2 receptor binding domain (RBD), induces robust and durable humoral, mucosal, and cellular immune responses of equivalent magnitude and quality in young and aged mice. This contrasted with the MF59-like intramuscular adjuvant, Addavax, which showed a marked decrease in immunogenicity with age. Robust antigen-specific IFNγ/IL-2/TNF-α was induced in both young and aged NE/IVT-immunized animals, which is significant as their reduced production is associated with suboptimal protective immunity in the elderly. These findings highlight the potential of adjuvanted mucosal vaccines for improving protection against COVID-19.
Language	English
Publishing date	2023-01-20
Publishing country	United States
Document type	Preprint
DOI	10.21203/rs.3.rs-2457013/v1
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Rescue of alveolar wall liquid secretion blocks fatal lung injury due to influenza-staphylococcal coinfection.

Tang, Stephanie / De Jesus, Ana Cassandra / Chavez, Deebly / Suthakaran, Sayahi / Moore, Sarah Kl / Suthakaran, Keshon / Homami, Sonya / Rathnasinghe, Raveen / May, Alison J / Schotsaert, Michael / Britto, Clemente J / Bhattacharya, Jahar / Hook, Jaime L

The Journal of clinical investigation

2023 Volume 133, Issue 19

Abstract: Secondary lung infection by inhaled Staphylococcus aureus (SA) is a common and lethal event for individuals infected with influenza A virus (IAV). How IAV disrupts host defense to promote SA infection in lung alveoli, where fatal lung injury occurs, is ... ...

Abstract	Secondary lung infection by inhaled Staphylococcus aureus (SA) is a common and lethal event for individuals infected with influenza A virus (IAV). How IAV disrupts host defense to promote SA infection in lung alveoli, where fatal lung injury occurs, is not known. We addressed this issue using real-time determinations of alveolar responses to IAV in live, intact, perfused lungs. Our findings show that IAV infection blocked defensive alveolar wall liquid (AWL) secretion and induced airspace liquid absorption, thereby reversing normal alveolar liquid dynamics and inhibiting alveolar clearance of inhaled SA. Loss of AWL secretion resulted from inhibition of the cystic fibrosis transmembrane conductance regulator (CFTR) ion channel in the alveolar epithelium, and airspace liquid absorption was caused by stimulation of the alveolar epithelial Na+ channel (ENaC). Loss of AWL secretion promoted alveolar stabilization of inhaled SA, but rescue of AWL secretion protected against alveolar SA stabilization and fatal SA-induced lung injury in IAV-infected mice. These findings reveal a central role for AWL secretion in alveolar defense against inhaled SA and identify AWL inhibition as a critical mechanism of IAV lung pathogenesis. AWL rescue may represent a new therapeutic approach for IAV-SA coinfection.
MeSH term(s)	Mice ; Animals ; Humans ; Influenza, Human/pathology ; Lung Injury/pathology ; Coinfection/pathology ; Pulmonary Alveoli/pathology ; Lung/pathology ; Influenza A virus
Language	English
Publishing date	2023-10-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	3067-3
ISSN	1558-8238 ; 0021-9738
ISSN (online)	1558-8238
ISSN	0021-9738
DOI	10.1172/JCI163402
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Article: The E484K mutation in the SARS-CoV-2 spike protein reduces but does not abolish neutralizing activity of human convalescent and post-vaccination sera.

Jangra, Sonia / Ye, Chengjin / Rathnasinghe, Raveen / Stadlbauer, Daniel / Krammer, Florian / Simon, Viviana / Martinez-Sobrido, Luis / García-Sastre, Adolfo / Schotsaert, Michael

medRxiv : the preprint server for health sciences

2021

Abstract: One year in the coronavirus disease 2019 (COVID-19) pandemic, the first vaccines are being rolled out under emergency use authorizations. It is of great concern that newly emerging variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ... ...

Abstract	One year in the coronavirus disease 2019 (COVID-19) pandemic, the first vaccines are being rolled out under emergency use authorizations. It is of great concern that newly emerging variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can escape antibody-mediated protection induced by previous infection or vaccination through mutations in the spike protein. The glutamate (E) to Lysine (K) substitution at position 484 (E484K) in the receptor binding domain (RBD) of the spike protein is present in the rapidly spreading variants of concern belonging to the B.1.351 and P.1 lineages. We performed
Language	English
Publishing date	2021-01-29
Publishing country	United States
Document type	Preprint
DOI	10.1101/2021.01.26.21250543
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: SARS-CoV-2 spike E484K mutation reduces antibody neutralisation.

Jangra, Sonia / Ye, Chengjin / Rathnasinghe, Raveen / Stadlbauer, Daniel / Krammer, Florian / Simon, Viviana / Martinez-Sobrido, Luis / García-Sastre, Adolfo / Schotsaert, Michael

The Lancet. Microbe

2021 Volume 2, Issue 7, Page(s) e283–e284

MeSH term(s)	Antibodies, Neutralizing/genetics ; COVID-19 ; Humans ; Mutation ; SARS-CoV-2/genetics ; Spike Glycoprotein, Coronavirus/genetics
Chemical Substances	Antibodies, Neutralizing ; Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2
Language	English
Publishing date	2021-04-07
Publishing country	England
Document type	Letter ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	2666-5247
ISSN (online)	2666-5247
DOI	10.1016/S2666-5247(21)00068-9
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Introduction of Two Prolines and Removal of the Polybasic Cleavage Site Lead to Higher Efficacy of a Recombinant Spike-Based SARS-CoV-2 Vaccine in the Mouse Model.

Amanat, Fatima / Strohmeier, Shirin / Rathnasinghe, Raveen / Schotsaert, Michael / Coughlan, Lynda / García-Sastre, Adolfo / Krammer, Florian

mBio

2021 Volume 12, Issue 2

Abstract: The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the prime target for vaccine development. The spike protein mediates both binding to host cells and membrane fusion and is also so far the only known ...

Abstract	The spike protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the prime target for vaccine development. The spike protein mediates both binding to host cells and membrane fusion and is also so far the only known viral target of neutralizing antibodies. Coronavirus spike proteins are large trimers that are relatively unstable, a feature that might be enhanced by the presence of a polybasic cleavage site in SARS-CoV-2 spike. Exchange of K986 and V987 for prolines has been shown to stabilize the trimers of SARS-CoV-1 and the Middle East respiratory syndrome coronavirus spike proteins. Here, we test multiple versions of a soluble spike protein for their immunogenicity and protective effect against SARS-CoV-2 challenge in a mouse model that transiently expresses human angiotensin-converting enzyme 2 via adenovirus transduction. Variants tested include spike proteins with a deleted polybasic cleavage site, proline mutations, or a combination thereof, besides the wild-type protein. While all versions of the protein were able to induce neutralizing antibodies, only the antigen with both a deleted cleavage site and the K986P and V987P (PP) mutations completely protected from challenge in this mouse model.
MeSH term(s)	Animals ; COVID-19 Vaccines/chemistry ; COVID-19 Vaccines/immunology ; COVID-19 Vaccines/therapeutic use ; Mice ; Mutation ; Proline/chemistry ; SARS-CoV-2/immunology ; Spike Glycoprotein, Coronavirus/chemistry
Chemical Substances	COVID-19 Vaccines ; Spike Glycoprotein, Coronavirus ; Proline (9DLQ4CIU6V)
Language	English
Publishing date	2021-03-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2557172-2
ISSN	2150-7511 ; 2161-2129
ISSN (online)	2150-7511
ISSN	2161-2129
DOI	10.1128/mBio.02648-20
Database	MEDical Literature Analysis and Retrieval System OnLINE

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