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Article ; Online: Quantification of Infectious SARS-CoV-2 by the 50% Tissue Culture Infectious Dose Endpoint Dilution Assay.

Bullen, C Korin / Davis, Stephanie L / Looney, Monika M

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2452, Page(s) 131–146

Abstract: A number of viral quantification methods are used to measure the concentration of infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). While the traditional plaque-based assay allows for direct enumeration of replication competent ... ...

Abstract	A number of viral quantification methods are used to measure the concentration of infectious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). While the traditional plaque-based assay allows for direct enumeration of replication competent lytic virions and remains the gold standard for the quantification of infectious virus, the 50% tissue culture infectious dose (TCID
MeSH term(s)	Animals ; Biological Assay/methods ; COVID-19 ; Communicable Diseases ; Cytopathogenic Effect, Viral ; SARS-CoV-2
Language	English
Publishing date	2022-05-12
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2111-0_9
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: MDA5 RNA-sensing pathway activation by Mycobacterium tuberculosis promotes innate immune subversion and pathogen survival.

Bullen, C Korin / Singh, Alok K / Krug, Stefanie / Lun, Shichun / Thakur, Preeti / Srikrishna, Geetha / Bishai, William R

JCI insight

2023 Volume 8, Issue 20

Abstract: Host cytosolic sensing of Mycobacterium tuberculosis (M. tuberculosis) RNA by the RIG-I-like receptor (RLR) family perturbs innate immune control within macrophages; however, a distinct role of MDA5, a member of the RLR family, in M. tuberculosis ... ...

Abstract	Host cytosolic sensing of Mycobacterium tuberculosis (M. tuberculosis) RNA by the RIG-I-like receptor (RLR) family perturbs innate immune control within macrophages; however, a distinct role of MDA5, a member of the RLR family, in M. tuberculosis pathogenesis has yet to be fully elucidated. To further define the role of MDA5 in M. tuberculosis pathogenesis, we evaluated M. tuberculosis intracellular growth and innate immune responses in WT and Mda5-/- macrophages. Transfection of M. tuberculosis RNA strongly induced proinflammatory cytokine production in WT macrophages, which was abrogated in Mda5-/- macrophages. M. tuberculosis infection in macrophages induced MDA5 protein expression, accompanied by an increase in MDA5 activation as assessed by multimer formation. IFN-γ-primed Mda5-/- macrophages effectively contained intracellular M. tuberculosis proliferation to a markedly greater degree than WT macrophages. Further comparisons of WT versus Mda5-/- macrophages revealed that during M. tuberculosis infection MDA5 contributed to IL-1β production and inflammasome activation and that loss of MDA5 led to a substantial increase in autophagy. In the mouse TB model, loss of MDA5 conferred host survival benefits with a concomitant reduction in M. tuberculosis bacillary burden. These data reveal that loss of MDA5 is host protective during M. tuberculosis infection in vitro and in vivo, suggesting that M. tuberculosis exploits MDA5 to subvert immune containment.
MeSH term(s)	Animals ; Mice ; Immunity, Innate ; Macrophages ; Mycobacterium tuberculosis ; RNA ; Tuberculosis
Chemical Substances	RNA (63231-63-0) ; Ifih1 protein, mouse (EC 3.6.1.-)
Language	English
Publishing date	2023-10-23
Publishing country	United States
Document type	Journal Article
ISSN	2379-3708
ISSN (online)	2379-3708
DOI	10.1172/jci.insight.166242
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Article ; Online: MDA5 RNA-sensing pathway activation by Mycobacterium tuberculosis promotes innate immune subversion and pathogen survival

C. Korin Bullen / Alok K. Singh / Stefanie Krug / Shichun Lun / Preeti Thakur / Geetha Srikrishna / William R. Bishai

JCI Insight, Vol 8, Iss

2023 Volume 20

Abstract: Host cytosolic sensing of Mycobacterium tuberculosis (M. tuberculosis) RNA by the RIG-I–like receptor (RLR) family perturbs innate immune control within macrophages; however, a distinct role of MDA5, a member of the RLR family, in M. tuberculosis ... ...

Abstract	Host cytosolic sensing of Mycobacterium tuberculosis (M. tuberculosis) RNA by the RIG-I–like receptor (RLR) family perturbs innate immune control within macrophages; however, a distinct role of MDA5, a member of the RLR family, in M. tuberculosis pathogenesis has yet to be fully elucidated. To further define the role of MDA5 in M. tuberculosis pathogenesis, we evaluated M. tuberculosis intracellular growth and innate immune responses in WT and Mda5–/– macrophages. Transfection of M. tuberculosis RNA strongly induced proinflammatory cytokine production in WT macrophages, which was abrogated in Mda5–/– macrophages. M. tuberculosis infection in macrophages induced MDA5 protein expression, accompanied by an increase in MDA5 activation as assessed by multimer formation. IFN-γ–primed Mda5–/– macrophages effectively contained intracellular M. tuberculosis proliferation to a markedly greater degree than WT macrophages. Further comparisons of WT versus Mda5–/– macrophages revealed that during M. tuberculosis infection MDA5 contributed to IL-1β production and inflammasome activation and that loss of MDA5 led to a substantial increase in autophagy. In the mouse TB model, loss of MDA5 conferred host survival benefits with a concomitant reduction in M. tuberculosis bacillary burden. These data reveal that loss of MDA5 is host protective during M. tuberculosis infection in vitro and in vivo, suggesting that M. tuberculosis exploits MDA5 to subvert immune containment.
Keywords	Immunology ; Infectious disease ; Medicine ; R
Language	English
Publishing date	2023-10-01T00:00:00Z
Publisher	American Society for Clinical investigation
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Genome editing of human embryos: to edit or not to edit, that is the question.

Chandrasegaran, Srinivasan / Bullen, C Korin / Carroll, Dana

The Journal of clinical investigation

2017 Volume 127, Issue 10, Page(s) 3588–3590

MeSH term(s)	Blastocyst ; Cardiomyopathy, Hypertrophic, Familial/genetics ; Cardiomyopathy, Hypertrophic, Familial/therapy ; Carrier Proteins/genetics ; Carrier Proteins/metabolism ; Gene Editing/methods ; Gene Editing/trends ; Germ-Line Mutation ; Humans
Chemical Substances	Carrier Proteins ; myosin-binding protein C
Language	English
Publishing date	2017-08-28
Publishing country	United States
Document type	Journal Article ; Editorial
ZDB-ID	3067-3
ISSN	1558-8238 ; 0021-9738
ISSN (online)	1558-8238
ISSN	0021-9738
DOI	10.1172/JCI96962
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Article ; Online: Nucleolin mediates SARS-CoV-2 replication and viral-induced apoptosis of host cells.

Merino, Vanessa F / Yan, Yu / Ordonez, Alvaro A / Bullen, C Korin / Lee, Albert / Saeki, Harumi / Ray, Krishanu / Huang, Tao / Jain, Sanjay K / Pomper, Martin G

Antiviral research

2023 Volume 211, Page(s) 105550

Abstract: Host-oriented antiviral therapeutics are promising treatment options to combat COVID-19 and its emerging variants. However, relatively little is known about the cellular proteins hijacked by SARS-CoV-2 for its replication. Here we show that SARS-CoV-2 ... ...

Abstract	Host-oriented antiviral therapeutics are promising treatment options to combat COVID-19 and its emerging variants. However, relatively little is known about the cellular proteins hijacked by SARS-CoV-2 for its replication. Here we show that SARS-CoV-2 induces expression and cytoplasmic translocation of the nucleolar protein, nucleolin (NCL). NCL interacts with SARS-CoV-2 viral proteins and co-localizes with N-protein in the nucleolus and in stress granules. Knockdown of NCL decreases the stress granule component G3BP1, viral replication and improved survival of infected host cells. NCL mediates viral-induced apoptosis and stress response via p53. SARS-CoV-2 increases NCL expression and nucleolar size and number in lungs of infected hamsters. Inhibition of NCL with the aptamer AS-1411 decreases viral replication and apoptosis of infected cells. These results suggest nucleolin as a suitable target for anti-COVID therapies.
MeSH term(s)	Humans ; SARS-CoV-2/metabolism ; DNA Helicases ; RNA Recognition Motif Proteins ; Poly-ADP-Ribose Binding Proteins ; RNA Helicases/metabolism ; COVID-19 ; Phosphoproteins/metabolism ; Apoptosis ; Virus Replication ; Nucleolin
Chemical Substances	DNA Helicases (EC 3.6.4.-) ; RNA Recognition Motif Proteins ; Poly-ADP-Ribose Binding Proteins ; RNA Helicases (EC 3.6.4.13) ; Phosphoproteins ; G3BP1 protein, human (EC 3.6.4.12)
Language	English
Publishing date	2023-02-03
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	306628-9
ISSN	1872-9096 ; 0166-3542
ISSN (online)	1872-9096
ISSN	0166-3542
DOI	10.1016/j.antiviral.2023.105550
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Zs.A 1627: Show issues

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Article ; Online: Relative and Quantitative Phosphoproteome Analysis of Macrophages in Response to Infection by Virulent and Avirulent

Choudhary, Eira / Bullen, C Korin / Goel, Renu / Singh, Alok Kumar / Praharaj, Monali / Thakur, Preeti / Dhiman, Rohan / Bishai, William R / Agarwal, Nisheeth

Journal of proteome research

2020 Volume 19, Issue 6, Page(s) 2316–2336

Abstract: Comparative phosphoproteomics ... ...

Abstract	Comparative phosphoproteomics of
MeSH term(s)	Humans ; Macrophages ; Mycobacterium bovis ; Mycobacterium tuberculosis ; RNA ; Tandem Mass Spectrometry
Chemical Substances	RNA (63231-63-0)
Language	English
Publishing date	2020-05-14
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2078618-9
ISSN	1535-3907 ; 1535-3893
ISSN (online)	1535-3907
ISSN	1535-3893
DOI	10.1021/acs.jproteome.9b00895
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Article ; Online: Intravenous BCG vaccination reduces SARS-CoV-2 severity and promotes extensive reprogramming of lung immune cells.

iScience

2023 Volume 26, Issue 10, Page(s) 107733

Abstract: Bacillus Calmette-Guérin (BCG) confers heterologous immune protection against viral infections and has been proposed as vaccine against SARS-CoV-2 (SCV2). Here, we tested intravenous BCG vaccination against COVID-19 using the golden Syrian hamster model. ...

Abstract	Bacillus Calmette-Guérin (BCG) confers heterologous immune protection against viral infections and has been proposed as vaccine against SARS-CoV-2 (SCV2). Here, we tested intravenous BCG vaccination against COVID-19 using the golden Syrian hamster model. BCG vaccination conferred a modest reduction on lung SCV2 viral load, bronchopneumonia scores, and weight loss, accompanied by a reversal of SCV2-mediated T cell lymphopenia, and reduced lung granulocytes. BCG uniquely recruited immunoglobulin-producing plasma cells to the lung suggesting accelerated local antibody production. BCG vaccination also recruited elevated levels of Th1, Th17, Treg, CTLs, and Tmem cells, with a transcriptional shift away from exhaustion markers and toward antigen presentation and repair. Similarly, BCG enhanced recruitment of alveolar macrophages and reduced key interstitial macrophage subsets, that show reduced IFN-associated gene expression. Our observations indicate that BCG vaccination protects against SCV2 immunopathology by promoting early lung immunoglobulin production and immunotolerizing transcriptional patterns among key myeloid and lymphoid populations.
Language	English
Publishing date	2023-08-24
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2023.107733
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Article: Dynamic single-cell RNA sequencing reveals BCG vaccination curtails SARS-CoV-2 induced disease severity and lung inflammation.

Singh, Alok K / Wang, Rulin / Lombardo, Kara A / Praharaj, Monali / Bullen, C Korin / Um, Peter / Davis, Stephanie / Komm, Oliver / Illei, Peter B / Ordonez, Alvaro A / Bahr, Melissa / Huang, Joy / Gupta, Anuj / Psoter, Kevin J / Jain, Sanjay K / Bivalacqua, Trinity J / Yegnasubramanian, Srinivasan / Bishai, William R

bioRxiv : the preprint server for biology

2022

Abstract: COVID-19 continues to exact a toll on human health despite the availability of several vaccines. Bacillus Calmette Guérin (BCG) has been shown to confer heterologous immune protection against viral infections including COVID-19 and has been proposed as ... ...

Abstract	COVID-19 continues to exact a toll on human health despite the availability of several vaccines. Bacillus Calmette Guérin (BCG) has been shown to confer heterologous immune protection against viral infections including COVID-19 and has been proposed as vaccine against SARS-CoV-2 (SCV2). Here we tested intravenous BCG vaccination against COVID-19 using the golden Syrian hamster model together with immune profiling and single cell RNA sequencing (scRNAseq). We observed that BCG reduced both lung SCV2 viral load and bronchopneumonia. This was accompanied by an increase in lung alveolar macrophages, a reversal of SCV2-mediated T cell lymphopenia, and reduced lung granulocytes. Single cell transcriptome profiling showed that BCG uniquely recruits immunoglobulin-producing plasma cells to the lung suggesting accelerated antibody production. BCG vaccination also recruited elevated levels of Th1, Th17, Treg, CTLs, and Tmem cells, and differentially expressed gene (DEG) analysis showed a transcriptional shift away from exhaustion markers and towards antigen presentation and repair. Similarly, BCG enhanced lung recruitment of alveolar macrophages and reduced key interstitial macrophage subsets, with both cell-types also showing reduced IFN-associated gene expression. Our observations indicate that BCG vaccination protects against SCV2 immunopathology by promoting early lung immunoglobulin production and immunotolerizing transcriptional patterns among key myeloid and lymphoid populations.
Language	English
Publishing date	2022-03-15
Publishing country	United States
Document type	Preprint
DOI	10.1101/2022.03.15.484018
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Article ; Online: Sulforaphane exhibits antiviral activity against pandemic SARS-CoV-2 and seasonal HCoV-OC43 coronaviruses in vitro and in mice.

Ordonez, Alvaro A / Bullen, C Korin / Villabona-Rueda, Andres F / Thompson, Elizabeth A / Turner, Mitchell L / Merino, Vanessa F / Yan, Yu / Kim, John / Davis, Stephanie L / Komm, Oliver / Powell, Jonathan D / D'Alessio, Franco R / Yolken, Robert H / Jain, Sanjay K / Jones-Brando, Lorraine

Communications biology

2022 Volume 5, Issue 1, Page(s) 242

Abstract: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the cause of coronavirus disease 2019 (COVID-19), has incited a global health crisis. Currently, there are limited therapeutic options for the prevention and treatment of SARS-CoV-2 infections. ...

Abstract	Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the cause of coronavirus disease 2019 (COVID-19), has incited a global health crisis. Currently, there are limited therapeutic options for the prevention and treatment of SARS-CoV-2 infections. We evaluated the antiviral activity of sulforaphane (SFN), the principal biologically active phytochemical derived from glucoraphanin, the naturally occurring precursor present in high concentrations in cruciferous vegetables. SFN inhibited in vitro replication of six strains of SARS-CoV-2, including Delta and Omicron, as well as that of the seasonal coronavirus HCoV-OC43. Further, SFN and remdesivir interacted synergistically to inhibit coronavirus infection in vitro. Prophylactic administration of SFN to K18-hACE2 mice prior to intranasal SARS-CoV-2 infection significantly decreased the viral load in the lungs and upper respiratory tract and reduced lung injury and pulmonary pathology compared to untreated infected mice. SFN treatment diminished immune cell activation in the lungs, including significantly lower recruitment of myeloid cells and a reduction in T cell activation and cytokine production. Our results suggest that SFN should be explored as a potential agent for the prevention or treatment of coronavirus infections.
MeSH term(s)	Adenosine Monophosphate/analogs & derivatives ; Adenosine Monophosphate/therapeutic use ; Alanine/analogs & derivatives ; Alanine/therapeutic use ; Animals ; Antiviral Agents/therapeutic use ; Caco-2 Cells ; Chlorocebus aethiops ; Common Cold/drug therapy ; Common Cold/virology ; Coronavirus Infections/drug therapy ; Coronavirus Infections/immunology ; Coronavirus Infections/virology ; Coronavirus OC43, Human ; Cytokines/immunology ; Drug Synergism ; Humans ; Isothiocyanates/therapeutic use ; Lung/immunology ; Lung/virology ; Macrophages, Alveolar/immunology ; Male ; Mice, Transgenic ; SARS-CoV-2 ; Spleen/immunology ; Sulfoxides/therapeutic use ; T-Lymphocytes/immunology ; Vero Cells ; Viral Load ; COVID-19 Drug Treatment
Chemical Substances	Antiviral Agents ; Cytokines ; Isothiocyanates ; Sulfoxides ; remdesivir (3QKI37EEHE) ; Adenosine Monophosphate (415SHH325A) ; sulforaphane (GA49J4310U) ; Alanine (OF5P57N2ZX)
Language	English
Publishing date	2022-03-18
Publishing country	England
Document type	Journal Article
ISSN	2399-3642
ISSN (online)	2399-3642
DOI	10.1038/s42003-022-03189-z
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Article: Sulforaphane exhibits in vitro and in vivo antiviral activity against pandemic SARS-CoV-2 and seasonal HCoV-OC43 coronaviruses.

Ordonez, Alvaro A / Bullen, C Korin / Villabona-Rueda, Andres F / Thompson, Elizabeth A / Turner, Mitchell L / Davis, Stephanie L / Komm, Oliver / Powell, Jonathan D / D'Alessio, Franco R / Yolken, Robert H / Jain, Sanjay K / Jones-Brando, Lorraine

bioRxiv : the preprint server for biology

2021

Abstract: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the cause of coronavirus disease 2019 (COVID-19), has incited a global health crisis. Currently, there are no orally available medications for prophylaxis for those exposed to SARS-CoV-2 and ... ...

Abstract	Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the cause of coronavirus disease 2019 (COVID-19), has incited a global health crisis. Currently, there are no orally available medications for prophylaxis for those exposed to SARS-CoV-2 and limited therapeutic options for those who develop COVID-19. We evaluated the antiviral activity of sulforaphane (SFN), a naturally occurring, orally available, well-tolerated, nutritional supplement present in high concentrations in cruciferous vegetables with limited side effects. SFN inhibited in vitro replication of four strains of SARS-CoV-2 as well as that of the seasonal coronavirus HCoV-OC43. Further, SFN and remdesivir interacted synergistically to inhibit coronavirus infection in vitro. Prophylactic administration of SFN to K18-hACE2 mice prior to intranasal SARS-CoV-2 infection significantly decreased the viral load in the lungs and upper respiratory tract and reduced lung injury and pulmonary pathology compared to untreated infected mice. SFN treatment diminished immune cell activation in the lungs, including significantly lower recruitment of myeloid cells and a reduction in T cell activation and cytokine production. Our results suggest that SFN is a promising treatment for prevention of coronavirus infection or treatment of early disease.
Language	English
Publishing date	2021-03-25
Publishing country	United States
Document type	Preprint
DOI	10.1101/2021.03.25.437060
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