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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.
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 StyleDennis 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 StyleDennis 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.
SARS-CoV-2 infection fatality ratios (IFR) remain controversially discussed with implications for political measures. The German county of Tirschenreuth suffered a severe SARS-CoV-2 outbreak in spring 2020, with particularly high case fatality ratio (CFR). To estimate seroprevalence, underreported infections, and IFR for the Tirschenreuth population aged ≥14 years in June/July 2020, we conducted a population-based study including home visits for the elderly, and analyzed 4203 participants for SARS-CoV-2 antibodies via three antibody tests. Latent class analysis yielded 8.6% standardized county-wide seroprevalence, a factor of underreported infections of 5.0, and 2.5% overall IFR. Seroprevalence was two-fold higher among medical workers and one third among current smokers with similar proportions of registered infections. While seroprevalence did not show an age-trend, the factor of underreported infections was 12.2 in the young versus 1.7 for ≥85-year-old. Age-specific IFRs were <0.5% below 60 years of age, 1.0% for age 60–69, and 13.2% for age 70+. Senior care homes accounted for 45% of COVID-19-related deaths, reflected by an IFR of 7.5% among individuals aged 70+ and an overall IFR of 1.4% when excluding senior care home residents from our computation. Our data underscore senior care home infections as key determinant of IFR additionally to age, insufficient targeted testing in the young, and the need for further investigations on behavioral or molecular causes of the fewer infections among current smokers.
Ralf Wagner; David Peterhoff; Stephanie Beileke; Felix Günther; Melanie Berr; Sebastian Einhauser; Anja Schütz; Hans Niller; Philipp Steininger; Antje Knöll; Matthias Tenbusch; Clara Maier; Klaus Korn; Klaus Stark; André Gessner; Ralph Burkhardt; Michael Kabesch; Holger Schedl; Helmut Küchenhoff; Annette Pfahlberg; Iris Heid; Olaf Gefeller; Klaus Überla. Estimates and Determinants of SARS-Cov-2 Seroprevalence and Infection Fatality Ratio Using Latent Class Analysis: The Population-Based Tirschenreuth Study in the Hardest-Hit German County in Spring 2020. Viruses 2021, 13, 1118 .
AMA StyleRalf Wagner, David Peterhoff, Stephanie Beileke, Felix Günther, Melanie Berr, Sebastian Einhauser, Anja Schütz, Hans Niller, Philipp Steininger, Antje Knöll, Matthias Tenbusch, Clara Maier, Klaus Korn, Klaus Stark, André Gessner, Ralph Burkhardt, Michael Kabesch, Holger Schedl, Helmut Küchenhoff, Annette Pfahlberg, Iris Heid, Olaf Gefeller, Klaus Überla. Estimates and Determinants of SARS-Cov-2 Seroprevalence and Infection Fatality Ratio Using Latent Class Analysis: The Population-Based Tirschenreuth Study in the Hardest-Hit German County in Spring 2020. Viruses. 2021; 13 (6):1118.
Chicago/Turabian StyleRalf Wagner; David Peterhoff; Stephanie Beileke; Felix Günther; Melanie Berr; Sebastian Einhauser; Anja Schütz; Hans Niller; Philipp Steininger; Antje Knöll; Matthias Tenbusch; Clara Maier; Klaus Korn; Klaus Stark; André Gessner; Ralph Burkhardt; Michael Kabesch; Holger Schedl; Helmut Küchenhoff; Annette Pfahlberg; Iris Heid; Olaf Gefeller; Klaus Überla. 2021. "Estimates and Determinants of SARS-Cov-2 Seroprevalence and Infection Fatality Ratio Using Latent Class Analysis: The Population-Based Tirschenreuth Study in the Hardest-Hit German County in Spring 2020." Viruses 13, no. 6: 1118.
The replication of viruses in secondary lymphoid organs guarantees sufficient amounts of pattern-recognition receptor ligands and antigens to activate the innate and adaptive immune system. Viruses with broad cell tropism usually replicate in lymphoid organs; however, whether a virus with a narrow tropism relies on replication in the secondary lymphoid organs to activate the immune system remains not well studied. In this study, we used the artificial intravenous route of infection to determine whether Influenza A virus (IAV) replication can occur in secondary lymphatic organs (SLO) and whether such replication correlates with innate immune activation. Indeed, we found that IAV replicates in secondary lymphatic tissue. IAV replication was dependent on the expression of Sialic acid residues in antigen-presenting cells and on the expression of the interferon-inhibitor UBP43 (Usp18). The replication of IAV correlated with innate immune activation, resulting in IAV eradication. The genetic deletion of Usp18 curbed IAV replication and limited innate immune activation. In conclusion, we found that IAV replicates in SLO, a mechanism which allows innate immune activation.
Sarah-Kim Friedrich; Rosa Schmitz; Michael Bergerhausen; Judith Lang; Vikas Duhan; Cornelia Hardt; Matthias Tenbusch; Marco Prinz; Kenichi Asano; Hilal Bhat; Thamer Hamdan; Philipp Lang; Karl Lang. Replication of Influenza A Virus in Secondary Lymphatic Tissue Contributes to Innate Immune Activation. Pathogens 2021, 10, 622 .
AMA StyleSarah-Kim Friedrich, Rosa Schmitz, Michael Bergerhausen, Judith Lang, Vikas Duhan, Cornelia Hardt, Matthias Tenbusch, Marco Prinz, Kenichi Asano, Hilal Bhat, Thamer Hamdan, Philipp Lang, Karl Lang. Replication of Influenza A Virus in Secondary Lymphatic Tissue Contributes to Innate Immune Activation. Pathogens. 2021; 10 (5):622.
Chicago/Turabian StyleSarah-Kim Friedrich; Rosa Schmitz; Michael Bergerhausen; Judith Lang; Vikas Duhan; Cornelia Hardt; Matthias Tenbusch; Marco Prinz; Kenichi Asano; Hilal Bhat; Thamer Hamdan; Philipp Lang; Karl Lang. 2021. "Replication of Influenza A Virus in Secondary Lymphatic Tissue Contributes to Innate Immune Activation." Pathogens 10, no. 5: 622.
Cell-cell fusion allows viruses to infect neighboring cells without the need to produce free virus and contributes to tissue damage by creating virus-infected syncytia. Our results demonstrate that the S2′ cleavage site is essential for activation by TMPRSS2 and unravel important differences between SARS-CoV and SARS-CoV-2, among those, greater dependence of SARS-CoV-2 on ACE2 expression and activation by metalloproteases for cell-cell fusion.
Bojan F. Hörnich; Anna K. Großkopf; Sarah Schlagowski; Matthias Tenbusch; Hannah Kleine-Weber; Frank Neipel; Christiane Stahl-Hennig; Alexander S. Hahn. SARS-CoV-2 and SARS-CoV Spike-Mediated Cell-Cell Fusion Differ in Their Requirements for Receptor Expression and Proteolytic Activation. Journal of Virology 2021, 95, 1 .
AMA StyleBojan F. Hörnich, Anna K. Großkopf, Sarah Schlagowski, Matthias Tenbusch, Hannah Kleine-Weber, Frank Neipel, Christiane Stahl-Hennig, Alexander S. Hahn. SARS-CoV-2 and SARS-CoV Spike-Mediated Cell-Cell Fusion Differ in Their Requirements for Receptor Expression and Proteolytic Activation. Journal of Virology. 2021; 95 (9):1.
Chicago/Turabian StyleBojan F. Hörnich; Anna K. Großkopf; Sarah Schlagowski; Matthias Tenbusch; Hannah Kleine-Weber; Frank Neipel; Christiane Stahl-Hennig; Alexander S. Hahn. 2021. "SARS-CoV-2 and SARS-CoV Spike-Mediated Cell-Cell Fusion Differ in Their Requirements for Receptor Expression and Proteolytic Activation." Journal of Virology 95, no. 9: 1.
SARS-CoV-2 infection fatality ratios (IFR) remain controversially discussed with implications for political measures, but the number of registered infections depends on testing strategies and deduced case fatality ratios (CFR) are poor proxies for IFR. The German county of Tirschenreuth suffered a severe SARS-CoV-2 outbreak in spring 2020 with particularly high CFR. To estimate seroprevalence, dark figure, and IFR for the Tirschenreuth population aged ≥14 years in June/July 2020 with misclassification error control, we conducted a population-based study, including home visits for elderly, and analyzed 4203 participants for SARS-CoV-2 antibodies via three antibody tests (64% of our random sample). Latent class analysis yielded 8.6% standardized county-wide seroprevalence, dark figure factor 5.0, and 2.5% overall IFR. Seroprevalence was two-fold higher among medical workers and one third among current smokers with similar proportions of registered infections. While seroprevalence did not show an age-trend, the dark figure was 12.2 in the young versus 1.7 for ≥85-year-old. Age-specific IFRs were <0.5% below 60 years of age, 1.0% for age 60-69, 13.2% for age 70+, confirming a previously reported age-model for IFR. Senior care homes accounted for 45% of COVID-19-related deaths, reflected by an IFR of 7.5% among individuals aged 70+ and an overall IFR of 1.4% when excluding senior care home residents from our computation. Our data underscore senior care home infections as key determinant of IFR additionally to age, insufficient targeted testing in the young, and the need for further investigations on behavioral or molecular causes of the fewer infections among current smokers.
Ralf Wagner; David Peterhoff; Stephanie Beileke; Felix Guenther; Melanie Berr; Sebastian Einhauser; Anja Schütz; Hans Helmut Niller; Philipp Steininger; Antje Knöll; Matthias Tenbusch; Clara Maier; Klaus Korn; Klaus J. Stark; Andre Gessner; Ralph Burkhardt; Michael Kabesch; Holger Schedl; Helmut Küchenhoff; Annette B. Pfahlberg; Iris M. Heid; Olaf Gefeller; Klaus Überla. Estimates and determinants of SARS-CoV-2 seroprevalence and infection fatality ratio using latent class analysis: the population-based Tirschenreuth study in the hardest-hit German county in spring 2020. 2021, 1 .
AMA StyleRalf Wagner, David Peterhoff, Stephanie Beileke, Felix Guenther, Melanie Berr, Sebastian Einhauser, Anja Schütz, Hans Helmut Niller, Philipp Steininger, Antje Knöll, Matthias Tenbusch, Clara Maier, Klaus Korn, Klaus J. Stark, Andre Gessner, Ralph Burkhardt, Michael Kabesch, Holger Schedl, Helmut Küchenhoff, Annette B. Pfahlberg, Iris M. Heid, Olaf Gefeller, Klaus Überla. Estimates and determinants of SARS-CoV-2 seroprevalence and infection fatality ratio using latent class analysis: the population-based Tirschenreuth study in the hardest-hit German county in spring 2020. . 2021; ():1.
Chicago/Turabian StyleRalf Wagner; David Peterhoff; Stephanie Beileke; Felix Guenther; Melanie Berr; Sebastian Einhauser; Anja Schütz; Hans Helmut Niller; Philipp Steininger; Antje Knöll; Matthias Tenbusch; Clara Maier; Klaus Korn; Klaus J. Stark; Andre Gessner; Ralph Burkhardt; Michael Kabesch; Holger Schedl; Helmut Küchenhoff; Annette B. Pfahlberg; Iris M. Heid; Olaf Gefeller; Klaus Überla. 2021. "Estimates and determinants of SARS-CoV-2 seroprevalence and infection fatality ratio using latent class analysis: the population-based Tirschenreuth study in the hardest-hit German county in spring 2020." , no. : 1.
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, only a limited number of validated serological assays were available in the early phase of the pandemic. Here, we evaluated a novel flow cytometric approach to assess spike-specific antibody responses.HEK 293T cells expressing SARS-CoV-2 spike protein in its natural confirmation on the surface were used to detect specific IgG and IgM antibody responses in patient sera by flow cytometry. A soluble angiotensin-converting-enzyme 2 (ACE-2) variant was developed as external standard to quantify spike-specific antibody responses on different assay platforms. 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. The external standard allowed robust quantification of antibody responses among 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. The flow cytometric assay also provides a blueprint for rapid development of serological tests to other emerging viral infections
Dennis Lapuente; Clara Maier; Pascal Irrgang; Julian Hübner; Antonia Sophia Peter; Markus Hoffmann; Armin Ensser; Katharina Ziegler; Thomas H. Winkler; Torsten Birkholz; Andreas E. Kremer; Philipp Steininger; Klaus Korn; Frank Neipel; Klaus Überla; Matthias Tenbusch. Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2. European Journal of Clinical Microbiology & Infectious Diseases 2020, 40, 751 -759.
AMA StyleDennis Lapuente, Clara Maier, Pascal Irrgang, Julian Hübner, Antonia Sophia Peter, Markus Hoffmann, Armin Ensser, Katharina Ziegler, Thomas H. Winkler, Torsten Birkholz, Andreas E. Kremer, Philipp Steininger, Klaus Korn, Frank Neipel, Klaus Überla, Matthias Tenbusch. Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2. European Journal of Clinical Microbiology & Infectious Diseases. 2020; 40 (4):751-759.
Chicago/Turabian StyleDennis Lapuente; Clara Maier; Pascal Irrgang; Julian Hübner; Antonia Sophia Peter; Markus Hoffmann; Armin Ensser; Katharina Ziegler; Thomas H. Winkler; Torsten Birkholz; Andreas E. Kremer; Philipp Steininger; Klaus Korn; Frank Neipel; Klaus Überla; Matthias Tenbusch. 2020. "Rapid response flow cytometric assay for the detection of antibody responses to SARS-CoV-2." European Journal of Clinical Microbiology & Infectious Diseases 40, no. 4: 751-759.
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.
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 StylePierre 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 StylePierre 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.
The severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) infects cells through interaction of its spike protein (SARS2-S) with Angiotensin-converting enzyme 2 (ACE2) and activation by proteases, in particular transmembrane protease serine 2 (TMPRSS2). Viruses can also spread through fusion of infected with uninfected cells. We compared the requirements of ACE2 expression, proteolytic activation, and the sensitivity to inhibitors for SARS2-S-mediated and SARS-CoV-S(SARS1-S)-mediated cell-cell fusion. SARS2-S-driven fusion was moderately increased by TMPRSS2 and strongly by ACE2, while SARS1-S-driven fusion was strongly increased by TMPRSS2 and less so by ACE2 expression. In contrast to SARS1-S, SARS2-S-mediated cell-cell fusion was efficiently activated by Batimastat-sensitive metalloproteases. Mutation of the S1/S2 proteolytic cleavage site reduced effector-target-cell fusion when ACE2 or TMPRSS2 were limiting and rendered SARS2-S-driven cell-cell fusion more dependent on TMPRSS2. When both ACE2 and TMPRSS2 were abundant, initial target-effector-cell fusion was unaltered compared to wt SARS2-S, but syncytia remained smaller. Mutation of the S2’ site specifically abrogated activation by TMPRSS2 for both cell-cell fusion and SARS2-S-driven pseudoparticle entry but still allowed for activation by metalloproteases for cell-cell fusion and by cathepsins for particle entry. Finally, we found that the TMPRSS2 inhibitor Bromhexine was unable to reduce TMPRSS2-activated cell-cell fusion by SARS1-S and SARS2-S as opposed to the inhibitor Camostat. Paradoxically, Bromhexine enhanced cell-cell fusion in the presence of TMPRSS2, while its metabolite Ambroxol exhibited inhibitory activity in some conditions. On Calu-3 lung cells, Ambroxol weakly inhibited SARS2-S-driven lentiviral pseudoparticle entry, and both substances exhibited a dose-dependent trend towards weak inhibition of authentic SARS-CoV-2. IMPORTANCE Cell-cell fusion allows the virus to infect neighboring cells without the need to produce free virus and contributes to tissue damage by creating virus-infected syncytia. Our results demonstrate that the S2’ cleavage site is essential for activation by TMPRSS2 and unravel important differences between SARS-CoV and SARS-CoV-2, among those greater dependence of SARS-CoV-2 on ACE2 expression and activation by metalloproteases for cell-cell fusion. Bromhexine, reportedly an inhibitor of TMPRSS2, is currently tested in clinical trials against coronavirus disease 2019. Our results indicate that Bromhexine enhances fusion in some conditions. We therefore caution against use of Bromhexine in higher dosage until its effects on SARS-CoV-2 spike activation are better understood. The related compound Ambroxol, which similarly to Bromhexine is clinically used as an expectorant, did not exhibit activating effects on cell-cell fusion. Both compounds exhibited weak inhibitory activity against SARS-CoV-2 infection at high concentrations, which might be clinically attainable for Ambroxol.
Bojan F. Hörnich; Anna K. Großkopf; Sarah Schlagowski; Matthias Tenbusch; Hannah Kleine-Weber; Frank Neipel; Christiane Stahl-Hennig; Alexander S. Hahn. SARS-CoV-2 and SARS-CoV spike-mediated cell-cell fusion differ in the requirements for receptor expression and proteolytic activation. 2020, 1 .
AMA StyleBojan F. Hörnich, Anna K. Großkopf, Sarah Schlagowski, Matthias Tenbusch, Hannah Kleine-Weber, Frank Neipel, Christiane Stahl-Hennig, Alexander S. Hahn. SARS-CoV-2 and SARS-CoV spike-mediated cell-cell fusion differ in the requirements for receptor expression and proteolytic activation. . 2020; ():1.
Chicago/Turabian StyleBojan F. Hörnich; Anna K. Großkopf; Sarah Schlagowski; Matthias Tenbusch; Hannah Kleine-Weber; Frank Neipel; Christiane Stahl-Hennig; Alexander S. Hahn. 2020. "SARS-CoV-2 and SARS-CoV spike-mediated cell-cell fusion differ in the requirements for receptor expression and proteolytic activation." , no. : 1.
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.
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 StyleDennis 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 StyleDennis 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.
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.
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 StyleLeonie 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 StyleLeonie 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.
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 StyleLeonie 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 StyleLeonie 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.
Combination immunotherapy (CIT) is currently applied as a treatment for different cancers and is proposed as a cure strategy for chronic viral infections. Whether such therapies are efficient during an acute infection remains elusive. To address this, inhibitory receptors were blocked and regulatory T cells depleted in acutely Friend retrovirus-infected mice. CIT resulted in a dramatic expansion of cytotoxic CD4+ and CD8+ T cells and a subsequent reduction in viral loads. Despite limited viral replication, mice developed fatal immunopathology after CIT. The pathology was most severe in the gastrointestinal tract and was mediated by granzyme B producing CD4+ and CD8+ T cells. A similar post-CIT pathology during acute Influenza virus infection of mice was observed, which could be prevented by vaccination. Melanoma patients who developed immune-related adverse events under immune checkpoint CIT also presented with expanded granzyme-expressing CD4+ and CD8+ T cell populations. Our data suggest that acute infections may induce immunopathology in patients treated with CIT, and that effective measures for infection prevention should be applied. Combination immunotherapy (CIT) directed against checkpoint mechanisms has been approved for the therapy of cancers and is proposed for the treatment of chronic infections. In cancer therapy patients often develop severe immunopathology under CIT. Here we show that acute viral infections (Friend retrovirus and Influenza virus) posed a significant threat during CIT in mice. The strong activation of cytotoxic T cells after an acute viral infection seemed to lack couterregulation during CIT, which resulted in lethal immunopathology. In case of an Influenza virus infection this could be prevented by vaccination prior to CIT. The expansion of CD4+ and CD8+ T cells expressing cytotoxic molecules were also observed in melanoma patients treated with two checkpoint blockers. Thus, these patients might also be at risk of developing severe immunopathology during an otherwise harmless acute viral infection. This should to be taken into account when cancer patients are undergoing CIT.
Paul David; Malgorzata Drabczyk-Pluta; Eva Pastille; Torben Knuschke; Tanja Werner; Nadine Honke; Dominik A. Megger; Ilseyar Akhmetzyanova; Namir Shaabani; Annette Eyking-Singer; Elke Cario; Olivia Kershaw; Achim D. Gruber; Matthias Tenbusch; Kirsten K. Dietze; Mirko Trilling; Jia Liu; Dirk Schadendorf; Hendrik Streeck; Karl Lang; Youhua Xie; Lisa Zimmer; Barbara Sitek; Annette Paschen; Astrid M. Westendorf; Ulf Dittmer; Gennadiy Zelinskyy. Combination immunotherapy with anti-PD-L1 antibody and depletion of regulatory T cells during acute viral infections results in improved virus control but lethal immunopathology. PLOS Pathogens 2020, 16, e1008340 .
AMA StylePaul David, Malgorzata Drabczyk-Pluta, Eva Pastille, Torben Knuschke, Tanja Werner, Nadine Honke, Dominik A. Megger, Ilseyar Akhmetzyanova, Namir Shaabani, Annette Eyking-Singer, Elke Cario, Olivia Kershaw, Achim D. Gruber, Matthias Tenbusch, Kirsten K. Dietze, Mirko Trilling, Jia Liu, Dirk Schadendorf, Hendrik Streeck, Karl Lang, Youhua Xie, Lisa Zimmer, Barbara Sitek, Annette Paschen, Astrid M. Westendorf, Ulf Dittmer, Gennadiy Zelinskyy. Combination immunotherapy with anti-PD-L1 antibody and depletion of regulatory T cells during acute viral infections results in improved virus control but lethal immunopathology. PLOS Pathogens. 2020; 16 (3):e1008340.
Chicago/Turabian StylePaul David; Malgorzata Drabczyk-Pluta; Eva Pastille; Torben Knuschke; Tanja Werner; Nadine Honke; Dominik A. Megger; Ilseyar Akhmetzyanova; Namir Shaabani; Annette Eyking-Singer; Elke Cario; Olivia Kershaw; Achim D. Gruber; Matthias Tenbusch; Kirsten K. Dietze; Mirko Trilling; Jia Liu; Dirk Schadendorf; Hendrik Streeck; Karl Lang; Youhua Xie; Lisa Zimmer; Barbara Sitek; Annette Paschen; Astrid M. Westendorf; Ulf Dittmer; Gennadiy Zelinskyy. 2020. "Combination immunotherapy with anti-PD-L1 antibody and depletion of regulatory T cells during acute viral infections results in improved virus control but lethal immunopathology." PLOS Pathogens 16, no. 3: e1008340.
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.
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 StylePierre 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 StylePierre 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.
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.
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 StyleRebecca 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 StyleRebecca 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.
Since preexisting immunity and enhanced infection rates in a clinical trial of an HIV vaccine have raised some concerns on adenovirus (Ad) serotype 5-based vaccines, we evaluated the subgroup D adenovirus serotype Ad19a for its suitability as novel viral vector vaccine against mucosal infections. In BALB/c mice, we compared the immunogenicity and efficacy of E1/E3-deleted Ad19a vectors encoding the influenza A virus (IAV)-derived antigens hemagglutinin (HA) and nucleoprotein (NP) to the most commonly used Ad5 vectors. The adenoviral vectors were applied intranasally and induced detectable antigen-specific T cell responses in the lung and in the spleen as well as robust antibody responses. A prior DNA immunization significantly improved the immunogenicity of both vectors and resulted in full protection against a lethal infection with a heterologous H3N2 virus. Nevertheless, the Ad5-based vectors were slightly superior in reducing viral replication in the lung which corresponded to higher NP-specific T cell responses measured in the lungs.
Dennis Lapuente; Zsolt Ruzsics; Christian Thirion; Matthias Tenbusch. Evaluation of adenovirus 19a as a novel vector for mucosal vaccination against influenza A viruses. Vaccine 2018, 36, 2712 -2720.
AMA StyleDennis Lapuente, Zsolt Ruzsics, Christian Thirion, Matthias Tenbusch. Evaluation of adenovirus 19a as a novel vector for mucosal vaccination against influenza A viruses. Vaccine. 2018; 36 (19):2712-2720.
Chicago/Turabian StyleDennis Lapuente; Zsolt Ruzsics; Christian Thirion; Matthias Tenbusch. 2018. "Evaluation of adenovirus 19a as a novel vector for mucosal vaccination against influenza A viruses." Vaccine 36, no. 19: 2712-2720.
A universal influenza vaccine must provide protection against antigenically divergent influenza viruses either through broadly neutralizing antibodies or cross-reactive T cells. Here, intranasal immunizations with recombinant adenoviral vectors (rAd) encoding hemagglutinin (HA) and nucleoprotein (NP) in combination with rAd-Interleukin-(IL)-1β or rAd-IL-18 were evaluated for their efficacy in BALB/c mice. Mucosal delivery of rAd-IL-1β enhanced HA-specific antibody responses including strain-specific neutralizing antibodies. Nevertheless, the beneficial effects on the local T cell responses were much more impressive reflected by increased numbers of CD103+CD69+ tissue-resident memory T cells (TRM). This increased immunogenicity translated into superior protection against infections with homologous and heterologous strains including H1N1, pH1N1, H3N2, and H7N7. Inhibition of the egress of circulating T cells out of the lymph nodes during the heterologous infection had no impact on the degree of protection underscoring the unique potential of TRM for the local containment of mucosal infections. The local co-expression of IL-1β and antigen lead to the activation of critical checkpoints in the formation of TRM including activation of epithelial cells, expression of chemokines and adhesion molecules, recruitment of lung-derived CD103+ DCs, and finally local TRM imprinting. Given the importance of TRM-mediated protection at mucosal barriers, this study has major implications for vaccine development.
D. Lapuente; M. Storcksdieck Genannt Bonsmann; A. Maaske; V. Stab; V. Heinecke; K. Watzstedt; R. Heß; A. M. Westendorf; W. Bayer; C. Ehrhardt; M. Tenbusch. IL-1β as mucosal vaccine adjuvant: the specific induction of tissue-resident memory T cells improves the heterosubtypic immunity against influenza A viruses. Mucosal Immunology 2018, 11, 1265 -1278.
AMA StyleD. Lapuente, M. Storcksdieck Genannt Bonsmann, A. Maaske, V. Stab, V. Heinecke, K. Watzstedt, R. Heß, A. M. Westendorf, W. Bayer, C. Ehrhardt, M. Tenbusch. IL-1β as mucosal vaccine adjuvant: the specific induction of tissue-resident memory T cells improves the heterosubtypic immunity against influenza A viruses. Mucosal Immunology. 2018; 11 (4):1265-1278.
Chicago/Turabian StyleD. Lapuente; M. Storcksdieck Genannt Bonsmann; A. Maaske; V. Stab; V. Heinecke; K. Watzstedt; R. Heß; A. M. Westendorf; W. Bayer; C. Ehrhardt; M. Tenbusch. 2018. "IL-1β as mucosal vaccine adjuvant: the specific induction of tissue-resident memory T cells improves the heterosubtypic immunity against influenza A viruses." Mucosal Immunology 11, no. 4: 1265-1278.
Retroviral envelope (Env) proteins are known to exhibit immunosuppressive properties, which become apparent not only in retroviral infections, but also in gene-based immunizations using retroviral immunogens, where envelope interferes with the induction of CD8+ T cell responses towards another, simultaneously or subsequently delivered, immunogen. In the Friend retrovirus mouse model, immunization with a plasmid encoding the Friend murine leukemia virus (F-MuLV) Leader-Gag protein resulted in induction of a strong GagL85–93-specific CD8+ T cell response, while the response was completely abrogated by co-immunization with an F-MuLV Env-encoding plasmid. In order to overcome this interference of retroviral envelope, we employed plasmids encoding the cytokines interleukin (IL) 1β, IL2, IL12, IL15, IL21, IL28A or granulocyte–macrophage colony-stimulating factor (GM-CSF) as genetic adjuvants. Co-application of plasmids encoding IL2, IL12, IL21, IL28A and especially GM-CSF rescued the induction of GagL85–93-specific CD8+ T cells in mice vaccinated with FV Leader-Gag and Env. Mice that were immunized with plasmids encoding Leader-Gag and Env and the cytokines IL1β, IL12, IL15, IL28A or GM-CSF, but not Leader-Gag and Env without any cytokine, showed significantly reduced viral loads upon a high-dose Friend virus challenge infection. Our data demonstrate the potency of cytokine-encoding vectors as adjuvants and immune modulators in composite vaccines for anti-retroviral immunization.
Nadine Bongard; Dennis Lapuente; Sonja Windmann; Ulf Dittmer; Matthias Tenbusch; Wibke Bayer. Interference of retroviral envelope with vaccine-induced CD8+ T cell responses is relieved by co-administration of cytokine-encoding vectors. Retrovirology 2017, 14, 1 -9.
AMA StyleNadine Bongard, Dennis Lapuente, Sonja Windmann, Ulf Dittmer, Matthias Tenbusch, Wibke Bayer. Interference of retroviral envelope with vaccine-induced CD8+ T cell responses is relieved by co-administration of cytokine-encoding vectors. Retrovirology. 2017; 14 (1):1-9.
Chicago/Turabian StyleNadine Bongard; Dennis Lapuente; Sonja Windmann; Ulf Dittmer; Matthias Tenbusch; Wibke Bayer. 2017. "Interference of retroviral envelope with vaccine-induced CD8+ T cell responses is relieved by co-administration of cytokine-encoding vectors." Retrovirology 14, no. 1: 1-9.