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Dennis Lapuente
Institute of Clinical and Molecular Virology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nürnberg

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Article
Published: 03 August 2021
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Several effective SARS-CoV-2 vaccines are currently in use, but in the light of waning immunity and the emergence of novel variants, effective boost modalities are needed in order to maintain or even increase immunity. Here we report that intranasal vaccinations with adenovirus 5 and 19a vectored vaccines following a systemic DNA or mRNA priming result in strong systemic and mucosal immunity in mice. In contrast to two intramuscular injections with an mRNA vaccine, the mucosal boost with adenoviral vectors induced high levels of IgA and tissue-resident memory T cells in the respiratory tract. Mucosal neutralization of virus variants of concern was also enhanced by the intranasal boosts. Importantly, priming with mRNA provoked a more comprehensive T cell response consisting of circulating and tissue-resident memory T cells after the boost, while a DNA priming induced mostly mucosal T cells. Concomitantly, the intranasal boost strategies provided protection against symptomatic disease. Therefore, a mucosal booster immunization after mRNA priming is a promising approach to establish mucosal immunity in addition to systemic responses.

ACS Style

Dennis Lapuente; Jana Fuchs; Jonas Willar; Ana V Antão; Valentina Eberlein; Nadja Uhlig; Leila Issmail; Anna Schmidt; Friederike Oltmanns; Antonia Sophia Peter; Sandra Mueller-Schmucker; Pascal Irrgang; Kirsten Fraedrich; Andrea Cara; Markus Hoffmann; Stefan Pöhlmann; Armin Ensser; Cordula Pertl; Torsten Willert; Christian Thirion; Thomas Grunwald; Klaus Überla; Matthias Tenbusch. Protective mucosal immunity against SARS-CoV-2 after heterologous systemic RNA-mucosal adenoviral vector immunization. 2021, 1 .

AMA Style

Dennis Lapuente, Jana Fuchs, Jonas Willar, Ana V Antão, Valentina Eberlein, Nadja Uhlig, Leila Issmail, Anna Schmidt, Friederike Oltmanns, Antonia Sophia Peter, Sandra Mueller-Schmucker, Pascal Irrgang, Kirsten Fraedrich, Andrea Cara, Markus Hoffmann, Stefan Pöhlmann, Armin Ensser, Cordula Pertl, Torsten Willert, Christian Thirion, Thomas Grunwald, Klaus Überla, Matthias Tenbusch. Protective mucosal immunity against SARS-CoV-2 after heterologous systemic RNA-mucosal adenoviral vector immunization. . 2021; ():1.

Chicago/Turabian Style

Dennis Lapuente; Jana Fuchs; Jonas Willar; Ana V Antão; Valentina Eberlein; Nadja Uhlig; Leila Issmail; Anna Schmidt; Friederike Oltmanns; Antonia Sophia Peter; Sandra Mueller-Schmucker; Pascal Irrgang; Kirsten Fraedrich; Andrea Cara; Markus Hoffmann; Stefan Pöhlmann; Armin Ensser; Cordula Pertl; Torsten Willert; Christian Thirion; Thomas Grunwald; Klaus Überla; Matthias Tenbusch. 2021. "Protective mucosal immunity against SARS-CoV-2 after heterologous systemic RNA-mucosal adenoviral vector immunization." , no. : 1.

Review
Published: 28 January 2021 in Viruses
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Current flu vaccines rely on the induction of strain-specific neutralizing antibodies, which leaves the population vulnerable to drifted seasonal or newly emerged pandemic strains. Therefore, universal flu vaccine approaches that induce broad immunity against conserved parts of influenza have top priority in research. Cross-reactive T cell responses, especially tissue-resident memory T cells in the respiratory tract, provide efficient heterologous immunity, and must therefore be a key component of universal flu vaccines. Here, we review recent findings about T cell-based flu immunity, with an emphasis on tissue-resident memory T cells in the respiratory tract of humans and different animal models. Furthermore, we provide an update on preclinical and clinical studies evaluating T cell-evoking flu vaccines, and discuss the implementation of T cell immunity in real-life vaccine policies.

ACS Style

Anna Schmidt; Dennis Lapuente. T Cell Immunity against Influenza: The Long Way from Animal Models Towards a Real-Life Universal Flu Vaccine. Viruses 2021, 13, 199 .

AMA Style

Anna Schmidt, Dennis Lapuente. T Cell Immunity against Influenza: The Long Way from Animal Models Towards a Real-Life Universal Flu Vaccine. Viruses. 2021; 13 (2):199.

Chicago/Turabian Style

Anna Schmidt; Dennis Lapuente. 2021. "T Cell Immunity against Influenza: The Long Way from Animal Models Towards a Real-Life Universal Flu Vaccine." Viruses 13, no. 2: 199.

Journal article
Published: 01 October 2020 in Vaccines
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Due to the low efficacy and the need for seasonal adaptation of currently licensed influenza A vaccines, the importance of alternative vaccination strategies is increasingly recognized. Considering that DNA vaccines can be rapidly manufactured and readily adapted with novel antigen sequences, genetic vaccination is a promising immunization platform. However, the applicability of different genetic adjuvants to this approach still represents a complex challenge. Immune checkpoints are a class of molecules involved in adaptive immune responses and germinal center reactions. In this study, we immunized mice by intramuscular electroporation with a DNA-vaccine encoding hemagglutinin (HA) and nucleoprotein (NP) of the influenza A virus. The DNA-vaccine was applied either alone or in combination with genetic adjuvants encoding the soluble ectodomains of programmed cell death protein-1 (sPD-1) or its ligand (sPD-L1). Co-administration of genetic checkpoint adjuvants did not significantly alter immune responses against NP. In contrast, sPD-1 co-electroporation elevated HA-specific CD4+ T cell responses, decreased regulatory CD4+ T cell pools, and modulated the IgG2a-biased HA antibody pattern towards an isotype-balanced IgG response with a trend to higher influenza neutralization in vitro. Taken together, our data demonstrate that a genetic DNA-adjuvant encoding soluble ectodomains of sPD-1 was able to modulate immune responses induced by a co-administered influenza DNA vaccine.

ACS Style

Pierre Tannig; Antonia Sophia Peter; Dennis Lapuente; Stephan Klessing; Anna Schmidt; Dominik Damm; Matthias Tenbusch; Klaus Überla; Vladimir Temchura. Genetic Co-Administration of Soluble PD-1 Ectodomains Modifies Immune Responses against Influenza A Virus Induced by DNA Vaccination. Vaccines 2020, 8, 570 .

AMA Style

Pierre Tannig, Antonia Sophia Peter, Dennis Lapuente, Stephan Klessing, Anna Schmidt, Dominik Damm, Matthias Tenbusch, Klaus Überla, Vladimir Temchura. Genetic Co-Administration of Soluble PD-1 Ectodomains Modifies Immune Responses against Influenza A Virus Induced by DNA Vaccination. Vaccines. 2020; 8 (4):570.

Chicago/Turabian Style

Pierre Tannig; Antonia Sophia Peter; Dennis Lapuente; Stephan Klessing; Anna Schmidt; Dominik Damm; Matthias Tenbusch; Klaus Überla; Vladimir Temchura. 2020. "Genetic Co-Administration of Soluble PD-1 Ectodomains Modifies Immune Responses against Influenza A Virus Induced by DNA Vaccination." Vaccines 8, no. 4: 570.

Other
Published: 13 May 2020
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SARS-CoV-2 has emerged as a previously unknown zoonotic coronavirus that spread worldwide causing a serious pandemic. While reliable nucleic acid-based diagnostic assays were rapidly available, there exists only a limited number of validated serological assays. Here, we evaluated a novel flow cytometric approach based on antigen-expressing HEK 293T cells to assess spike-specific IgG and IgM antibody responses. Analyses of 201 pre-COVID-19 sera proved a high assay specificity in comparison to commercially available CLIA and ELISA systems, while also revealing the highest sensitivity in specimens from PCR-confirmed SARS-CoV-2 infected patients. Additionally, a soluble Angiotensin-Converting-Enzyme 2 (ACE-2) variant was established as external standard to quantify spike-specific antibody responses on different assay platforms. In conclusion, our newly established flow cytometric assay allows sensitive and quantitative detection of SARS-CoV-2-specific antibodies, which can be easily adopted in different laboratories and does not rely on external supply of assay kits.

ACS Style

Dennis Lapuente; Clara Maier; Pascal Irrgang; Julian Huebner; Sophia Antonia Peter; Markus Hoffmann; Armin Ensser; Katharina Ziegler; Thomas H. Winkler; Thorsten Birkholz; Andreas E. Kremer; Philipp Steininger; Klaus Korn; Frank Neipel; Klaus Ueberla; Matthias Tenbusch. Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2. 2020, 1 .

AMA Style

Dennis Lapuente, Clara Maier, Pascal Irrgang, Julian Huebner, Sophia Antonia Peter, Markus Hoffmann, Armin Ensser, Katharina Ziegler, Thomas H. Winkler, Thorsten Birkholz, Andreas E. Kremer, Philipp Steininger, Klaus Korn, Frank Neipel, Klaus Ueberla, Matthias Tenbusch. Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2. . 2020; ():1.

Chicago/Turabian Style

Dennis Lapuente; Clara Maier; Pascal Irrgang; Julian Huebner; Sophia Antonia Peter; Markus Hoffmann; Armin Ensser; Katharina Ziegler; Thomas H. Winkler; Thorsten Birkholz; Andreas E. Kremer; Philipp Steininger; Klaus Korn; Frank Neipel; Klaus Ueberla; Matthias Tenbusch. 2020. "Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2." , no. : 1.

Short communication
Published: 23 April 2020 in Biochemical and Biophysical Research Communications
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The Coxsackie- and adenovirus receptor (CAR) mediates homophilic cell-cell contacts and susceptibility to both human pathogenic viruses through its membrane-distal immunoglobulin domain. In the present study, we screened five missense variants of the human CAR gene for their influence on adenovector or Coxsackievirus entry into Chinese hamster ovary cells. The CAR variants facilitated virus internalisation to a similar extent as wild type CAR. This underlines CAR’s presumed invariance and essential physiological role in embryogenesis.

ACS Style

Leonie Herrmann; Adrian Filip; Dennis Lapuente; Matthias Tenbusch; Karsten Niehaus; Volker Rudolph; Martin Farr. Naturally occurring variants in the transmembrane and cytoplasmic domains of the human Coxsackie- and adenovirus receptor have no impact on virus internalisation. Biochemical and Biophysical Research Communications 2020, 527, 401 -405.

AMA Style

Leonie Herrmann, Adrian Filip, Dennis Lapuente, Matthias Tenbusch, Karsten Niehaus, Volker Rudolph, Martin Farr. Naturally occurring variants in the transmembrane and cytoplasmic domains of the human Coxsackie- and adenovirus receptor have no impact on virus internalisation. Biochemical and Biophysical Research Communications. 2020; 527 (2):401-405.

Chicago/Turabian Style

Leonie Herrmann; Adrian Filip; Dennis Lapuente; Matthias Tenbusch; Karsten Niehaus; Volker Rudolph; Martin Farr. 2020. "Naturally occurring variants in the transmembrane and cytoplasmic domains of the human Coxsackie- and adenovirus receptor have no impact on virus internalisation." Biochemical and Biophysical Research Communications 527, no. 2: 401-405.

Journal article
Published: 22 April 2020
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ACS Style

Leonie Herrmann; Adrian Filip; Dennis Lapuente; Matthias Tenbusch; Karsten Niehaus; Volker Rudolph; Martin Farr. Naturally occurring variants in the transmembrane and cytoplasmic domains of the human Coxsackie- and adenovirus receptor have no impact on virus internalisation. 2020, 1 .

AMA Style

Leonie Herrmann, Adrian Filip, Dennis Lapuente, Matthias Tenbusch, Karsten Niehaus, Volker Rudolph, Martin Farr. Naturally occurring variants in the transmembrane and cytoplasmic domains of the human Coxsackie- and adenovirus receptor have no impact on virus internalisation. . 2020; ():1.

Chicago/Turabian Style

Leonie Herrmann; Adrian Filip; Dennis Lapuente; Matthias Tenbusch; Karsten Niehaus; Volker Rudolph; Martin Farr. 2020. "Naturally occurring variants in the transmembrane and cytoplasmic domains of the human Coxsackie- and adenovirus receptor have no impact on virus internalisation." , no. : 1.

Research article
Published: 03 April 2020 in PLOS ONE
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In respect to the heterogeneity among influenza A virus strains and the shortcomings of current vaccination programs, there is a huge interest in the development of alternative vaccines that provide a broader and more long-lasting protection. Gene-based approaches are considered as promising candidates for such flu vaccines. In our study, innate signalling molecules from the RIG-I and the NALP3 pathways were evaluated as genetic adjuvants in intramuscular DNA immunizations. Plasmids encoding a constitutive active form of RIG-I (cRIG-I), IPS-1, IL-1β, or IL-18 were co-administered with plasmids encoding the hemagglutinin and nucleoprotein derived from H1N1/Puerto Rico/8/1934 via electroporation in BALB/c mice. Immunogenicity was analysed in detail and efficacy was demonstrated in homologous and heterologous influenza challenge experiments. Although the biological activities of the adjuvants have been confirmed by in vitro reporter assays, their single or combined inclusion in the vaccine did not result in superior vaccine efficacy. With the exception of significantly increased levels of antigen-specific IgG1 after the co-administration of IL-1β, there were only minor alterations concerning the immunogenicity. Since DNA electroporation alone induced substantial inflammation at the injection site, as demonstrated in this study using Mx2-Luc reporter mice, it might override the adjuvants´ contribution to the inflammatory microenvironment and thereby minimizes the influence on the immunogenicity. Taken together, the DNA immunization was protective against subsequent challenge infections but could not be further improved by the genetic adjuvants analysed in this study.

ACS Style

Dennis Lapuente; Viktoria Stab; Michael Storcksdieck Genannt Bonsmann; Andre Maaske; Mario Köster; Han Xiao; Christina Ehrhardt; Matthias Tenbusch. Innate signalling molecules as genetic adjuvants do not alter the efficacy of a DNA-based influenza A vaccine. PLOS ONE 2020, 15, e0231138 .

AMA Style

Dennis Lapuente, Viktoria Stab, Michael Storcksdieck Genannt Bonsmann, Andre Maaske, Mario Köster, Han Xiao, Christina Ehrhardt, Matthias Tenbusch. Innate signalling molecules as genetic adjuvants do not alter the efficacy of a DNA-based influenza A vaccine. PLOS ONE. 2020; 15 (4):e0231138.

Chicago/Turabian Style

Dennis Lapuente; Viktoria Stab; Michael Storcksdieck Genannt Bonsmann; Andre Maaske; Mario Köster; Han Xiao; Christina Ehrhardt; Matthias Tenbusch. 2020. "Innate signalling molecules as genetic adjuvants do not alter the efficacy of a DNA-based influenza A vaccine." PLOS ONE 15, no. 4: e0231138.

Journal article
Published: 14 January 2020 in Vaccines
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The importance of a balanced TH1/TH2 humoral immune response against the HIV-1 envelope protein (Env) for antibody-mediated HIV-1 control is increasingly recognized. However, there is no defined vaccination strategy to raise it. Since immune checkpoints are involved in the induction of adoptive immunity and their inhibitors (monoclonal antibodies) are licensed for cancer therapy, we investigated the effect of checkpoint blockade after HIV-1 genetic vaccination on enhancement and modulation of antiviral antibody responses. By intraperitoneal administration of checkpoint antibodies in mice we observed an induction of anti-drug antibodies which may interfere with immunomodulation by checkpoint inhibitors. Therefore, we blocked immune checkpoints locally by co-electroporation of DNA vaccines encoding the active soluble ectodomains of programmed cell death protein-1 (PD-1) or its ligand (PD-L1), respectively. Plasmid-encoded immune checkpoints did not elicit a detectable antibody response, suggesting no interference with their immunomodulatory effects. Co-electroporation of a HIV-1 DNA vaccine formulation with soluble PD-L1 ectodomain increased HIV-1 Env-specific TH1 CD4 T cell and IgG2a antibody responses. The overall antibody response was hereby shifted towards a more TH1/TH2 balanced subtype pattern. These findings indicate that co-electroporation of soluble checkpoint ectodomains together with DNA-based vaccines has modulatory effects on vaccine-induced immune responses that could improve vaccine efficacies.

ACS Style

Pierre Tannig; Antonia Sophia Peter; Dennis Lapuente; Stephan Klessing; Dominik Damm; Matthias Tenbusch; Klaus Überla; Vladimir Temchura. Modulation of Vaccine-Induced HIV-1-Specific Immune Responses by Co-Electroporation of PD-L1 Encoding DNA. Vaccines 2020, 8, 27 .

AMA Style

Pierre Tannig, Antonia Sophia Peter, Dennis Lapuente, Stephan Klessing, Dominik Damm, Matthias Tenbusch, Klaus Überla, Vladimir Temchura. Modulation of Vaccine-Induced HIV-1-Specific Immune Responses by Co-Electroporation of PD-L1 Encoding DNA. Vaccines. 2020; 8 (1):27.

Chicago/Turabian Style

Pierre Tannig; Antonia Sophia Peter; Dennis Lapuente; Stephan Klessing; Dominik Damm; Matthias Tenbusch; Klaus Überla; Vladimir Temchura. 2020. "Modulation of Vaccine-Induced HIV-1-Specific Immune Responses by Co-Electroporation of PD-L1 Encoding DNA." Vaccines 8, no. 1: 27.

Journal article
Published: 13 February 2019 in Viruses
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The envelope protein (Env) is the only surface protein of the human immunodeficiency virus (HIV) and as such the exclusive target for protective antibody responses. Experimental evidences from mouse models suggest a modulating property of Env to steer antibody class switching towards the less effective antibody subclass IgG1 accompanied with strong TH2 helper responses. By simple physical linkage we were able to imprint this bias, exemplified by a low IgG2a/IgG1 ratio of antigen-specific antibodies, onto an unrelated antigen, namely the HIV capsid protein p24. Here, our results indicate the glycan moiety of Env as the responsible immune modulating activity. Firstly, in Card9−/− mice lacking specific C-Type lectin responsiveness, DNA immunization significantly increased the IgG2a/IgG1 ratio for the Env-specific antibodies while the antibody response against the F-protein of the respiratory syncytial virus (RSV) serving as control antigen remained unchanged. Secondly, sequential shortening of the Env encoding sequence revealed the C2V3 domain as responsible for the strong IgG1 responses and TH2 cytokine production. Removing all potential N-glycosylation sites from the C2V3 domain by site-specific mutagenesis reversed the vaccine-induced immune response towards a Th1-dominated T-cell response and a balanced IgG2a/IgG1 ratio. Accordingly, the stretch of oligomannose glycans in the C2V3 domain of Env might mediate a specific uptake and/or signaling modus in antigen presenting cells by involving interaction with an as yet unknown C-type lectin receptor. Our results contribute to a deeper understanding of the impact of Env glycosylation on HIV antigen-specific immune responses, which will further support HIV vaccine development.

ACS Style

Rebecca Heß; Michael Storcksdieck Genannt Bonsmann; Dennis Lapuente; Andre Maaske; Carsten Kirschning; Jürgen Ruland; Bernd Lepenies; Drew Hannaman; Matthias Tenbusch; Klaus Überla. Glycosylation of HIV Env Impacts IgG Subtype Responses to Vaccination. Viruses 2019, 11, 153 .

AMA Style

Rebecca Heß, Michael Storcksdieck Genannt Bonsmann, Dennis Lapuente, Andre Maaske, Carsten Kirschning, Jürgen Ruland, Bernd Lepenies, Drew Hannaman, Matthias Tenbusch, Klaus Überla. Glycosylation of HIV Env Impacts IgG Subtype Responses to Vaccination. Viruses. 2019; 11 (2):153.

Chicago/Turabian Style

Rebecca Heß; Michael Storcksdieck Genannt Bonsmann; Dennis Lapuente; Andre Maaske; Carsten Kirschning; Jürgen Ruland; Bernd Lepenies; Drew Hannaman; Matthias Tenbusch; Klaus Überla. 2019. "Glycosylation of HIV Env Impacts IgG Subtype Responses to Vaccination." Viruses 11, no. 2: 153.