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Article ; Online: Single-cell suspension preparation from murine organs following

Ndeupen, Sonia / Qin, Zhen / Igyártó, Botond Z

STAR protocols

2022 Volume 3, Issue 2, Page(s) 101350

Abstract: ... for downstream analyses. For complete details on the use and execution of this protocol, please refer to Ndeupen ...

Abstract	We describe a protocol to study inflammatory responses triggered by the mRNA-lipid nanoparticle (LNP) vaccine formulations in skin, muscle, and lung and the adaptive immune responses induced in the draining lymph nodes. Here, we will present how to deliver these reagents through intradermal, intramuscular, and intranasal routes, generating single-cell suspensions from the inoculated and target organs for downstream analyses. For complete details on the use and execution of this protocol, please refer to Ndeupen et al. (2021) and (2022).
MeSH term(s)	Animals ; Liposomes ; Mice ; Nanoparticles/adverse effects ; RNA, Messenger/genetics ; Suspensions
Chemical Substances	Lipid Nanoparticles ; Liposomes ; RNA, Messenger ; Suspensions
Language	English
Publishing date	2022-05-18
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2022.101350
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Future considerations for the mRNA-lipid nanoparticle vaccine platform.

Igyártó, Botond Z / Jacobsen, Sonya / Ndeupen, Sonia

Current opinion in virology

2021 Volume 48, Page(s) 65–72

Abstract: Vaccines based on mRNA-containing lipid nanoparticles (LNPs) pioneered by Katalin Karikó and Drew Weissman at the University of Pennsylvania are a promising new vaccine platform used by two of the leading vaccines against coronavirus disease in 2019 ( ... ...

Abstract	Vaccines based on mRNA-containing lipid nanoparticles (LNPs) pioneered by Katalin Karikó and Drew Weissman at the University of Pennsylvania are a promising new vaccine platform used by two of the leading vaccines against coronavirus disease in 2019 (COVID-19). However, there are many questions regarding their mechanism of action in humans that remain unanswered. Here we consider the immunological features of LNP components and off-target effects of the mRNA, both of which could increase the risk of side effects. We suggest ways to mitigate these potential risks by harnessing dendritic cell (DC) biology.
MeSH term(s)	COVID-19/prevention & control ; COVID-19 Vaccines/immunology ; Dendritic Cells/immunology ; Humans ; Immunization/methods ; Lipids/administration & dosage ; Lipids/immunology ; Nanoparticles/administration & dosage ; Nanoparticles/metabolism ; RNA, Messenger/administration & dosage ; RNA, Messenger/immunology ; Vaccines/classification
Chemical Substances	COVID-19 Vaccines ; Lipids ; RNA, Messenger ; Vaccines
Language	English
Publishing date	2021-04-24
Publishing country	Netherlands
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	2611378-8
ISSN	1879-6265 ; 1879-6257
ISSN (online)	1879-6265
ISSN	1879-6257
DOI	10.1016/j.coviro.2021.03.008
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Single-cell suspension preparation from murine organs following in vivo mRNA-LNP exposure

Sonia Ndeupen / Zhen Qin / Botond Z. Igyártó

STAR Protocols, Vol 3, Iss 2, Pp 101350- (2022)

2022

Abstract: ... for downstream analyses.For complete details on the use and execution of this protocol, please refer to Ndeupen ...

Abstract	Summary: We describe a protocol to study inflammatory responses triggered by the mRNA-lipid nanoparticle (LNP) vaccine formulations in skin, muscle, and lung and the adaptive immune responses induced in the draining lymph nodes. Here, we will present how to deliver these reagents through intradermal, intramuscular, and intranasal routes, generating single-cell suspensions from the inoculated and target organs for downstream analyses.For complete details on the use and execution of this protocol, please refer to Ndeupen et al. (2021) and (2022).
Keywords	Health Sciences ; Immunology ; Science (General) ; Q1-390
Language	English
Publishing date	2022-06-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: The mRNA-LNP platform's lipid nanoparticle component used in preclinical vaccine studies is highly inflammatory.

Ndeupen, Sonia / Qin, Zhen / Jacobsen, Sonya / Bouteau, Aurélie / Estanbouli, Henri / Igyártó, Botond Z

iScience

2021 Volume 24, Issue 12, Page(s) 103479

Abstract: Vaccines based on mRNA-containing lipid nanoparticles (LNPs) are a promising new platform used by two leading vaccines against COVID-19. Clinical trials and ongoing vaccinations present with varying degrees of protection levels and side effects. However, ...

Abstract	Vaccines based on mRNA-containing lipid nanoparticles (LNPs) are a promising new platform used by two leading vaccines against COVID-19. Clinical trials and ongoing vaccinations present with varying degrees of protection levels and side effects. However, the drivers of the reported side effects remain poorly defined. Here we present evidence that Acuitas' LNPs used in preclinical nucleoside-modified mRNA vaccine studies are highly inflammatory in mice. Intradermal and intramuscular injection of these LNPs led to rapid and robust inflammatory responses, characterized by massive neutrophil infiltration, activation of diverse inflammatory pathways, and production of various inflammatory cytokines and chemokines. The same dose of LNP delivered intranasally led to similar inflammatory responses in the lung and resulted in a high mortality rate, with mechanism unresolved. Thus, the mRNA-LNP platforms' potency in supporting the induction of adaptive immune responses and the observed side effects may stem from the LNPs' highly inflammatory nature.
Language	English
Publishing date	2021-11-20
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2021.103479
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: The mRNA-LNP platform's lipid nanoparticle component used in preclinical vaccine studies is highly inflammatory.

Ndeupen, Sonia / Qin, Zhen / Jacobsen, Sonya / Estanbouli, Henri / Bouteau, Aurélie / Igyártó, Botond Z

bioRxiv : the preprint server for biology

2021

Abstract: Vaccines based on mRNA-containing lipid nanoparticles (LNPs) are a promising new platform used by two leading vaccines against coronavirus disease in 2019 (COVID-19). Clinical trials and ongoing vaccinations present with very high protection levels and ... ...

Abstract	Vaccines based on mRNA-containing lipid nanoparticles (LNPs) are a promising new platform used by two leading vaccines against coronavirus disease in 2019 (COVID-19). Clinical trials and ongoing vaccinations present with very high protection levels and varying degrees of side effects. However, the nature of the reported side effects remains poorly defined. Here we present evidence that LNPs used in many preclinical studies are highly inflammatory in mice. Intradermal injection of these LNPs led to rapid and robust inflammatory responses, characterized by massive neutrophil infiltration, activation of diverse inflammatory pathways, and production of various inflammatory cytokines and chemokines. The same dose of LNP delivered intranasally led to similar inflammatory responses in the lung and resulted in a high mortality rate. In summary, here we show that the LNPs used for many preclinical studies are highly inflammatory. Thus, their potent adjuvant activity and reported superiority comparing to other adjuvants in supporting the induction of adaptive immune responses likely stem from their inflammatory nature. Furthermore, the preclinical LNPs are similar to the ones used for human vaccines, which could also explain the observed side effects in humans using this platform.
Language	English
Publishing date	2021-07-23
Publishing country	United States
Document type	Preprint
DOI	10.1101/2021.03.04.430128
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: The mRNA-LNP platform's lipid nanoparticle component used in preclinical vaccine studies is highly inflammatory

Sonia Ndeupen / Zhen Qin / Sonya Jacobsen / Aurélie Bouteau / Henri Estanbouli / Botond Z. Igyártó

iScience, Vol 24, Iss 12, Pp 103479- (2021)

2021

Abstract: Summary: Vaccines based on mRNA-containing lipid nanoparticles (LNPs) are a promising new platform used by two leading vaccines against COVID-19. Clinical trials and ongoing vaccinations present with varying degrees of protection levels and side effects. ...

Abstract	Summary: Vaccines based on mRNA-containing lipid nanoparticles (LNPs) are a promising new platform used by two leading vaccines against COVID-19. Clinical trials and ongoing vaccinations present with varying degrees of protection levels and side effects. However, the drivers of the reported side effects remain poorly defined. Here we present evidence that Acuitas' LNPs used in preclinical nucleoside-modified mRNA vaccine studies are highly inflammatory in mice. Intradermal and intramuscular injection of these LNPs led to rapid and robust inflammatory responses, characterized by massive neutrophil infiltration, activation of diverse inflammatory pathways, and production of various inflammatory cytokines and chemokines. The same dose of LNP delivered intranasally led to similar inflammatory responses in the lung and resulted in a high mortality rate, with mechanism unresolved. Thus, the mRNA-LNP platforms' potency in supporting the induction of adaptive immune responses and the observed side effects may stem from the LNPs' highly inflammatory nature.
Keywords	Biological sciences ; Immunology ; Biotechnology ; Science ; Q
Subject code	610
Language	English
Publishing date	2021-12-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Langerhans cells and cDC1s play redundant roles in mRNA-LNP induced protective anti-influenza and anti-SARS-CoV-2 responses.

Ndeupen, Sonia / Bouteau, Aurélie / Herbst, Christopher / Qin, Zhen / Hutchins, Zachary / Kurup, Drishya / Diba, Leila Zabihi / Igyártó, Botond Z

bioRxiv : the preprint server for biology

2021

Abstract: Nucleoside modified mRNA combined with Acuitas Therapeutics' lipid nanoparticles (LNP) have been shown to support robust humoral immune responses in many preclinical animal vaccine studies and later in humans with the SARS-CoV-2 vaccination. We recently ... ...

Abstract	Nucleoside modified mRNA combined with Acuitas Therapeutics' lipid nanoparticles (LNP) have been shown to support robust humoral immune responses in many preclinical animal vaccine studies and later in humans with the SARS-CoV-2 vaccination. We recently showed that this platform is highly inflammatory due to the LNPs' ionizable lipid component. The inflammatory property is key to support the development of potent humoral immune responses. However, the mechanism by which this platform drives T follicular helper cells (Tfh) and humoral immune responses remains unknown. Here we show that lack of Langerhans cells or cDC1s neither significantly affected the induction of PR8 HA and SARS-CoV-2 RBD-specific Tfh cells and humoral immune responses, nor susceptibility towards the lethal challenge of influenza and SARS-CoV-2. However, the combined deletion of these two DC subsets led to a significant decrease in the induction of PR8 HA and SARS-CoV-2 RBD-specific Tfh cell and humoral immune responses. Despite these observed defects, the still high antibody titers were sufficient to confer protection towards lethal viral challenges. We further found that IL-6, but not neutrophils, was required to generate Tfh cells and antibody responses. In summary, here we bring evidence that the mRNA-LNP platform can support protective adaptive immune responses in the absence of specific DC subsets through an IL-6 dependent and neutrophil independent mechanism.
Language	English
Publishing date	2021-08-02
Publishing country	United States
Document type	Preprint
DOI	10.1101/2021.08.01.454662
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Langerhans cells and cDC1s play redundant roles in mRNA-LNP induced protective anti-influenza and anti-SARS-CoV-2 immune responses.

Ndeupen, Sonia / Bouteau, Aurélie / Herbst, Christopher / Qin, Zhen / Jacobsen, Sonya / Powers, Nicholas E / Hutchins, Zachary / Kurup, Drishya / Diba, Leila Zabihi / Watson, Megan / Ramage, Holly / Igyártó, Botond Z

PLoS pathogens

2022 Volume 18, Issue 1, Page(s) e1010255

Abstract: Nucleoside modified mRNA combined with Acuitas Therapeutics' lipid nanoparticles (LNPs) has been shown to support robust humoral immune responses in many preclinical animal vaccine studies and later in humans with the SARS-CoV-2 vaccination. We recently ... ...

Abstract	Nucleoside modified mRNA combined with Acuitas Therapeutics' lipid nanoparticles (LNPs) has been shown to support robust humoral immune responses in many preclinical animal vaccine studies and later in humans with the SARS-CoV-2 vaccination. We recently showed that this platform is highly inflammatory due to the LNPs' ionizable lipid component. The inflammatory property is key to support the development of potent humoral immune responses. However, the mechanism by which this platform drives T follicular helper (Tfh) cells and humoral immune responses remains unknown. Here we show that lack of Langerhans cells or cDC1s neither significantly affected the induction of PR8 HA and SARS-CoV-2 RBD-specific Tfh cells and humoral immune responses, nor susceptibility towards the lethal challenge of influenza and SARS-CoV-2. However, the combined deletion of these two DC subsets led to a significant decrease in the induction of PR8 HA and SARS-CoV-2 RBD-specific Tfh cell and humoral immune responses. Despite these observed defects, these mice remained protected from lethal influenza and SARS-CoV-2 challenges. We further found that IL-6, unlike neutrophils, was required to generate normal Tfh cells and antibody responses, but not for protection from influenza challenge. In summary, here we bring evidence that the mRNA-LNP platform can support the induction of protective immune responses in the absence of certain innate immune cells and cytokines.
MeSH term(s)	Animals ; COVID-19/immunology ; COVID-19 Vaccines/immunology ; Dendritic Cells/immunology ; Influenza Vaccines/immunology ; Langerhans Cells/immunology ; Liposomes/immunology ; Mice ; Nanoparticles ; Orthomyxoviridae Infections/immunology ; SARS-CoV-2/immunology ; Vaccines, Synthetic/immunology ; mRNA Vaccines/immunology
Chemical Substances	COVID-19 Vaccines ; Influenza Vaccines ; Lipid Nanoparticles ; Liposomes ; Vaccines, Synthetic ; mRNA Vaccines
Language	English
Publishing date	2022-01-24
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2205412-1
ISSN	1553-7374 ; 1553-7374
ISSN (online)	1553-7374
ISSN	1553-7374
DOI	10.1371/journal.ppat.1010255
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Langerhans cells and cDC1s play redundant roles in mRNA-LNP induced protective anti-influenza and anti-SARS-CoV-2 immune responses.

Sonia Ndeupen / Aurélie Bouteau / Christopher Herbst / Zhen Qin / Sonya Jacobsen / Nicholas E Powers / Zachary Hutchins / Drishya Kurup / Leila Zabihi Diba / Megan Watson / Holly Ramage / Botond Z Igyártó

PLoS Pathogens, Vol 18, Iss 1, p e

2022 Volume 1010255

Abstract	Nucleoside modified mRNA combined with Acuitas Therapeutics' lipid nanoparticles (LNPs) has been shown to support robust humoral immune responses in many preclinical animal vaccine studies and later in humans with the SARS-CoV-2 vaccination. We recently showed that this platform is highly inflammatory due to the LNPs' ionizable lipid component. The inflammatory property is key to support the development of potent humoral immune responses. However, the mechanism by which this platform drives T follicular helper (Tfh) cells and humoral immune responses remains unknown. Here we show that lack of Langerhans cells or cDC1s neither significantly affected the induction of PR8 HA and SARS-CoV-2 RBD-specific Tfh cells and humoral immune responses, nor susceptibility towards the lethal challenge of influenza and SARS-CoV-2. However, the combined deletion of these two DC subsets led to a significant decrease in the induction of PR8 HA and SARS-CoV-2 RBD-specific Tfh cell and humoral immune responses. Despite these observed defects, these mice remained protected from lethal influenza and SARS-CoV-2 challenges. We further found that IL-6, unlike neutrophils, was required to generate normal Tfh cells and antibody responses, but not for protection from influenza challenge. In summary, here we bring evidence that the mRNA-LNP platform can support the induction of protective immune responses in the absence of certain innate immune cells and cytokines.
Keywords	Immunologic diseases. Allergy ; RC581-607 ; Biology (General) ; QH301-705.5
Subject code	570
Language	English
Publishing date	2022-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Langerhans cells and cDC1s play redundant roles in mRNA-LNP induced protective anti-influenza and anti-SARS-CoV-2 responses

Ndeupen, Sonia / Bouteau, Aurelie / Herbst, Christopher / Qin, Zhen / Hutchins, Zachary / Kurup, Drishya / Diba, Leila Zabihi / Igyarto, Botond

bioRxiv

Abstract: Nucleoside modified mRNA combined with Acuitas Therapeutics9 lipid nanoparticles (LNP) have been shown to support robust humoral immune responses in many preclinical animal vaccine studies and later in humans with the SARS-CoV-2 vaccination. We recently ... ...

Abstract	Nucleoside modified mRNA combined with Acuitas Therapeutics9 lipid nanoparticles (LNP) have been shown to support robust humoral immune responses in many preclinical animal vaccine studies and later in humans with the SARS-CoV-2 vaccination. We recently showed that this platform is highly inflammatory due to the LNPs9 ionizable lipid component. The inflammatory property is key to support the development of potent humoral immune responses. However, the mechanism by which this platform drives T follicular helper cells (Tfh) and humoral immune responses remains unknown. Here we show that lack of Langerhans cells or cDC1s neither significantly affected the induction of PR8 HA and SARS-CoV-2 RBD-specific Tfh cells and humoral immune responses, nor susceptibility towards the lethal challenge of influenza and SARS-CoV-2. However, the combined deletion of these two DC subsets led to a significant decrease in the induction of PR8 HA and SARS-CoV-2 RBD-specific Tfh cell and humoral immune responses. Despite these observed defects, the still high antibody titers were sufficient to confer protection towards lethal viral challenges. We further found that IL-6, but not neutrophils, was required to generate Tfh cells and antibody responses. In summary, here we bring evidence that the mRNA-LNP platform can support protective adaptive immune responses in the absence of specific DC subsets through an IL-6 dependent and neutrophil independent mechanism.
Keywords	covid19
Language	English
Publishing date	2021-08-02
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2021.08.01.454662
Database	COVID19

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