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Currently, human infections with the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) are accelerating the ongoing spread of the pandemic. Several innovative types of vaccines have already been developed, whereas effective options of antiviral treatments still await a scientific implementation. The development of novel anti-SARS-CoV-2 drug candidates demands skillful strategies and analysis systems. Promising results have been achieved with first generation direct-acting antivirals targeting the viral polymerase RdRp or the protease 3CLpro. Such recently approved or investigational drugs like remdesivir and GC376 represent a basis for further development and optimization. Here, we establish a multi-readout assay (MRA) system that enables the antiviral assessment and mechanistic characterization of novel test compounds, drug repurposing and combination treatments. Our SARS-CoV-2-specific MRA combines the quantitative measurement of several parameters of virus infection, such as the intracellular production of proteins and genomes, enzymatic activities and virion release, as well as the use of reporter systems. In this regard, the antiviral efficacy of remdesivir and GC376 has been investigated in human Caco-2 cells. The readouts included the use of spike- and double-strand RNA-specific monoclonal antibodies for in-cell fluorescence imaging, a newly generated recombinant SARS-CoV-2 reporter virus d6YFP, the novel 3CLpro-based FRET CFP::YFP and the previously reported FlipGFP reporter assays, as well as viral genome-specific RT-qPCR. The data produced by our MRA confirm the high antiviral potency of these two drugs in vitro. Combined, this MRA approach may be applied for broader analyses of SARS-CoV-2-specific antivirals, including compound screenings and the characterization of selected drug candidates.
Friedrich Hahn; Sigrun Häge; Alexandra Herrmann; Christina Wangen; Jintawee Kicuntod; Doris Jungnickl; Julia Tillmanns; Regina Müller; Kirsten Fraedrich; Klaus Überla; Hella Kohlhof; Armin Ensser; Manfred Marschall. Methodological Development of a Multi-Readout Assay for the Assessment of Antiviral Drugs against SARS-CoV-2. Pathogens 2021, 10, 1076 .
AMA StyleFriedrich Hahn, Sigrun Häge, Alexandra Herrmann, Christina Wangen, Jintawee Kicuntod, Doris Jungnickl, Julia Tillmanns, Regina Müller, Kirsten Fraedrich, Klaus Überla, Hella Kohlhof, Armin Ensser, Manfred Marschall. Methodological Development of a Multi-Readout Assay for the Assessment of Antiviral Drugs against SARS-CoV-2. Pathogens. 2021; 10 (9):1076.
Chicago/Turabian StyleFriedrich Hahn; Sigrun Häge; Alexandra Herrmann; Christina Wangen; Jintawee Kicuntod; Doris Jungnickl; Julia Tillmanns; Regina Müller; Kirsten Fraedrich; Klaus Überla; Hella Kohlhof; Armin Ensser; Manfred Marschall. 2021. "Methodological Development of a Multi-Readout Assay for the Assessment of Antiviral Drugs against SARS-CoV-2." Pathogens 10, no. 9: 1076.
Bernhard Fleckenstein, Professor Emeritus and former Chair of the Institute of Clinical and Molecular Virology at the Friedrich-Alexander-University Erlangen-Nuremberg (FAU), Erlangen, Germany, passed away on May 4th, 2021.…
Andrea K. Thoma-Kress; Ronald C. Desrosiers; Michaela U. Gack; Jae U. Jung; Klaus Überla; Frank Kirchhoff. Biographical Feature: Bernhard Fleckenstein. Journal of Virology 2021, 95, JVI0089621 .
AMA StyleAndrea K. Thoma-Kress, Ronald C. Desrosiers, Michaela U. Gack, Jae U. Jung, Klaus Überla, Frank Kirchhoff. Biographical Feature: Bernhard Fleckenstein. Journal of Virology. 2021; 95 (17):JVI0089621.
Chicago/Turabian StyleAndrea K. Thoma-Kress; Ronald C. Desrosiers; Michaela U. Gack; Jae U. Jung; Klaus Überla; Frank Kirchhoff. 2021. "Biographical Feature: Bernhard Fleckenstein." Journal of Virology 95, no. 17: JVI0089621.
TRIANNI mice carry an entire set of human immunoglobulin V region gene segments and are a powerful tool to rapidly isolate human monoclonal antibodies. After immunizing these mice with DNA encoding the spike protein of SARS-CoV-2 and boosting with spike protein, we identified 29 hybridoma antibodies that reacted with the SARS-CoV-2 spike protein. Nine antibodies neutralize SARS-CoV-2 infection at IC50 values in the subnanomolar range. ELISA-binding studies and DNA sequence analyses revealed one cluster of three clonally related neutralizing antibodies that target the receptor-binding domain and compete with the cellular receptor hACE2. A second cluster of six clonally related neutralizing antibodies bind to the N-terminal domain of the spike protein without competing with the binding of hACE2 or cluster 1 antibodies. SARS-CoV-2 mutants selected for resistance to an antibody from one cluster are still neutralized by an antibody from the other cluster. Antibodies from both clusters markedly reduced viral spread in mice transgenic for human ACE2 and protected the animals from SARS-CoV-2-induced weight loss. The two clusters of potent non-competing SARS-CoV-2 neutralizing antibodies provide potential candidates for therapy and prophylaxis of COVID-19. The study further supports transgenic animals with a human immunoglobulin gene repertoire as a powerful platform in pandemic preparedness initiatives. This article is protected by copyright. All rights reserved
Antonia Sophia Peter; Edith Roth; Sebastian R. Schulz; Kirsten Fraedrich; Tobit Steinmetz; Dominik Damm; Manuela Hauke; Elie Richel; Sandra Mueller‐Schmucker; Katharina Habenicht; Valentina Eberlein; Leila Issmail; Nadja Uhlig; Simon Dolles; Eva Grüner; David Peterhoff; Sandra Ciesek; Markus Hoffmann; Stefan Pöhlmann; Paul F. McKay; Robin J. Shattock; Roman Wölfel; Eileen Socher; Ralf Wagner; Jutta Eichler; Heinrich Sticht; Wolfgang Schuh; Frank Neipel; Armin Ensser; Dirk Mielenz; Matthias Tenbusch; Thomas H. Winkler; Thomas Grunwald; Klaus Überla; Hans‐Martin Jäck. A pair of non‐competing neutralizing human monoclonal antibodies protecting from disease in a SARS‐CoV‐2 infection model. European Journal of Immunology 2021, 1 .
AMA StyleAntonia Sophia Peter, Edith Roth, Sebastian R. Schulz, Kirsten Fraedrich, Tobit Steinmetz, Dominik Damm, Manuela Hauke, Elie Richel, Sandra Mueller‐Schmucker, Katharina Habenicht, Valentina Eberlein, Leila Issmail, Nadja Uhlig, Simon Dolles, Eva Grüner, David Peterhoff, Sandra Ciesek, Markus Hoffmann, Stefan Pöhlmann, Paul F. McKay, Robin J. Shattock, Roman Wölfel, Eileen Socher, Ralf Wagner, Jutta Eichler, Heinrich Sticht, Wolfgang Schuh, Frank Neipel, Armin Ensser, Dirk Mielenz, Matthias Tenbusch, Thomas H. Winkler, Thomas Grunwald, Klaus Überla, Hans‐Martin Jäck. A pair of non‐competing neutralizing human monoclonal antibodies protecting from disease in a SARS‐CoV‐2 infection model. European Journal of Immunology. 2021; ():1.
Chicago/Turabian StyleAntonia Sophia Peter; Edith Roth; Sebastian R. Schulz; Kirsten Fraedrich; Tobit Steinmetz; Dominik Damm; Manuela Hauke; Elie Richel; Sandra Mueller‐Schmucker; Katharina Habenicht; Valentina Eberlein; Leila Issmail; Nadja Uhlig; Simon Dolles; Eva Grüner; David Peterhoff; Sandra Ciesek; Markus Hoffmann; Stefan Pöhlmann; Paul F. McKay; Robin J. Shattock; Roman Wölfel; Eileen Socher; Ralf Wagner; Jutta Eichler; Heinrich Sticht; Wolfgang Schuh; Frank Neipel; Armin Ensser; Dirk Mielenz; Matthias Tenbusch; Thomas H. Winkler; Thomas Grunwald; Klaus Überla; Hans‐Martin Jäck. 2021. "A pair of non‐competing neutralizing human monoclonal antibodies protecting from disease in a SARS‐CoV‐2 infection model." European Journal of Immunology , 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.
Alternative splicing and the expression of intron-containing mRNAs is one hallmark of HIV gene expression. To facilitate the otherwise hampered nuclear export of non-fully processed mRNAs, HIV encodes the Rev protein, which recognizes its intronic response element and fuels the HIV RNAs into the CRM-1-dependent nuclear protein export pathway. Both alternative splicing and Rev-dependency are regulated by the primary HIV RNA sequence. Here, we show that these processes are extremely sensitive to sequence alterations in the 5’coding region of the HIV genomic RNA. Increasing the GC content by insertion of either GFP or silent mutations activates a cryptic splice donor site in gag, entirely deregulates the viral splicing pattern, and lowers infectivity. Interestingly, an adaptation of the inserted GFP sequence toward an HIV-like nucleotide bias reversed these phenotypes completely. Of note, the adaptation yielded completely different primary sequences although encoding the same amino acids. Thus, the phenotypes solely depend on the nucleotide composition of the two GFP versions. This is a strong indication of an HIV-specific mRNP code in the 5′ gag region wherein the primary RNA sequence bias creates motifs for RNA-binding proteins and controls the fate of the HIV-RNA in terms of viral gene expression and infectivity.
Bastian Grewe; Carolin Vogt; Theresa Horstkötter; Bettina Tippler; Han Xiao; Bianca Müller; Klaus Überla; Ralf Wagner; Benedikt Asbach; Jens Bohne. The HIV 5′ Gag Region Displays a Specific Nucleotide Bias Regulating Viral Splicing and Infectivity. Viruses 2021, 13, 997 .
AMA StyleBastian Grewe, Carolin Vogt, Theresa Horstkötter, Bettina Tippler, Han Xiao, Bianca Müller, Klaus Überla, Ralf Wagner, Benedikt Asbach, Jens Bohne. The HIV 5′ Gag Region Displays a Specific Nucleotide Bias Regulating Viral Splicing and Infectivity. Viruses. 2021; 13 (6):997.
Chicago/Turabian StyleBastian Grewe; Carolin Vogt; Theresa Horstkötter; Bettina Tippler; Han Xiao; Bianca Müller; Klaus Überla; Ralf Wagner; Benedikt Asbach; Jens Bohne. 2021. "The HIV 5′ Gag Region Displays a Specific Nucleotide Bias Regulating Viral Splicing and Infectivity." Viruses 13, no. 6: 997.
TRIANNI mice carry an entire set of human immunoglobulin V region gene segments and are a powerful tool to rapidly generate human monoclonal antibodies. After immunizing these mice against the spike protein of SARS-CoV-2, we identified 29 hybridoma antibodies that reacted with the SARS-CoV-2 spike protein. Nine antibodies neutralized SARS-CoV-2 infection at IC50 values in the subnanomolar range. ELISA-binding studies and DNA sequence analyses revealed one cluster of clonally related neutralizing antibodies that target the receptor-binding domain and compete with the cellular receptor hACE2. A second cluster of neutralizing antibodies binds to the N-terminal domain of the spike protein without competing with the binding of hACE2 or cluster 1 antibodies. SARS-CoV-2 mutants selected for resistance to an antibody from one cluster are still neutralized by an antibody from the other cluster. Antibodies from both clusters markedly reduced viral spread in mice transgenic for human ACE2 and protected the animals from SARS-CoV-2 induced weight loss. Thus, we report two clusters of potent non-competing SARS-CoV-2 neutralizing antibodies providing potential candidates for therapy and prophylaxis of COVID-19. The study further supports the use of transgenic animals with human immunoglobulin gene repertoires in pandemic preparedness initiatives.
Antonia Sophia Peter; Edith Roth; Sebastian R. Schulz; Kirsten Fraedrich; Tobit Steinmetz; Dominik Damm; Manuela Hauke; Elie Richel; Sandra Mueller-Schmucker; Katharina Habenicht; Valentina Eberlein; Leila Issmail; Nadja Uhlig; Simon Dolles; Eva Gruner; David Peterhoff; Sandra Ciesek; Markus Hoffmann; Stefan Pohlmann; Paul F. McKay; Robin J. Shattock; Roman Wolfel; Ralf Wagner; Jutta Eichler; Wolfgang Schuh; Frank Neipel; Armin Ensser; Dirk Mielenz; Matthias Tenbusch; Thomas H. Winkler; Thomas Grunwald; Klaus Uberla; Hans-Martin Jack. A pair of non-competing neutralizing human monoclonal antibodies protecting from disease in a SARS-CoV-2 infection model. 2021, 1 .
AMA StyleAntonia Sophia Peter, Edith Roth, Sebastian R. Schulz, Kirsten Fraedrich, Tobit Steinmetz, Dominik Damm, Manuela Hauke, Elie Richel, Sandra Mueller-Schmucker, Katharina Habenicht, Valentina Eberlein, Leila Issmail, Nadja Uhlig, Simon Dolles, Eva Gruner, David Peterhoff, Sandra Ciesek, Markus Hoffmann, Stefan Pohlmann, Paul F. McKay, Robin J. Shattock, Roman Wolfel, Ralf Wagner, Jutta Eichler, Wolfgang Schuh, Frank Neipel, Armin Ensser, Dirk Mielenz, Matthias Tenbusch, Thomas H. Winkler, Thomas Grunwald, Klaus Uberla, Hans-Martin Jack. A pair of non-competing neutralizing human monoclonal antibodies protecting from disease in a SARS-CoV-2 infection model. . 2021; ():1.
Chicago/Turabian StyleAntonia Sophia Peter; Edith Roth; Sebastian R. Schulz; Kirsten Fraedrich; Tobit Steinmetz; Dominik Damm; Manuela Hauke; Elie Richel; Sandra Mueller-Schmucker; Katharina Habenicht; Valentina Eberlein; Leila Issmail; Nadja Uhlig; Simon Dolles; Eva Gruner; David Peterhoff; Sandra Ciesek; Markus Hoffmann; Stefan Pohlmann; Paul F. McKay; Robin J. Shattock; Roman Wolfel; Ralf Wagner; Jutta Eichler; Wolfgang Schuh; Frank Neipel; Armin Ensser; Dirk Mielenz; Matthias Tenbusch; Thomas H. Winkler; Thomas Grunwald; Klaus Uberla; Hans-Martin Jack. 2021. "A pair of non-competing neutralizing human monoclonal antibodies protecting from disease in a SARS-CoV-2 infection model." , no. : 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.
The HIV-1 Rev protein is a nuclear export factor for unspliced and incompletely spliced HIV-1 RNAs. Without Rev, these intron-retaining RNAs are trapped in the nucleus. A genome-wide screen identified nine proteins of the spliceosome, which all enhanced expression from the HIV-1 unspliced RNA after CRISPR/Cas knockdown. Depletion of DHX38, WDR70, and four proteins of the Prp19-associated complex (ISY1, BUD31, XAB2, and CRNKL1) resulted in a more than 20-fold enhancement of unspliced HIV-1 RNA levels in the cytoplasm. Targeting of CRNKL1, DHX38, and BUD31 affected nuclear export efficiencies of the HIV-1 unspliced RNA to a much larger extent than splicing. Transcriptomic analyses further revealed that CRNKL1 also suppresses cytoplasmic levels of a subset of cellular mRNAs, including some with selectively retained introns. Thus, CRNKL1-dependent nuclear retention is a novel cellular mechanism for the regulation of cytoplasmic levels of intron-retaining HIV-1 mRNAs, which HIV-1 may have harnessed to direct its complex splicing pattern. IMPORTANCE To regulate its complex splicing pattern, HIV-1 uses the adaptor protein Rev to shuttle unspliced or partially spliced mRNA from the nucleus to the cytoplasm. In the absence of Rev, these RNAs are retained in the nucleus, but it is unclear why. Here we identify cellular proteins whose depletion enhances cytoplasmic levels of the HIV-1 unspliced RNA. Depletion of one of them, CRNKL1, also increases cytoplasmic levels of a subset of intron-retaining cellular mRNA, suggesting that CRNKL1-dependent nuclear retention may be a basic cellular mechanism exploited by HIV-1.
Han Xiao; Emanuel Wyler; Miha Milek; Bastian Grewe; Philipp Kirchner; Arif Ekici; Ana Beatriz Oliveira Villela Silva; Doris Jungnickl; Florian Full; Marco Thomas; Markus Landthaler; Armin Ensser; Klaus Überla. CRNKL1 Is a Highly Selective Regulator of Intron-Retaining HIV-1 and Cellular mRNAs. mBio 2021, 12, 1 .
AMA StyleHan Xiao, Emanuel Wyler, Miha Milek, Bastian Grewe, Philipp Kirchner, Arif Ekici, Ana Beatriz Oliveira Villela Silva, Doris Jungnickl, Florian Full, Marco Thomas, Markus Landthaler, Armin Ensser, Klaus Überla. CRNKL1 Is a Highly Selective Regulator of Intron-Retaining HIV-1 and Cellular mRNAs. mBio. 2021; 12 (1):1.
Chicago/Turabian StyleHan Xiao; Emanuel Wyler; Miha Milek; Bastian Grewe; Philipp Kirchner; Arif Ekici; Ana Beatriz Oliveira Villela Silva; Doris Jungnickl; Florian Full; Marco Thomas; Markus Landthaler; Armin Ensser; Klaus Überla. 2021. "CRNKL1 Is a Highly Selective Regulator of Intron-Retaining HIV-1 and Cellular mRNAs." mBio 12, no. 1: 1.
The ongoing pandemic spread of the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) demands skillful strategies for novel drug development, drug repurposing and cotreatments, in particular focusing on existing candidates of host-directed antivirals (HDAs). The developmental drug IMU-838, currently being investigated in a phase 2b trial in patients suffering from autoimmune diseases, represents an inhibitor of human dihydroorotate dehydrogenase (DHODH) with a recently proven antiviral activity in vitro and in vivo. Here, we established an analysis system for assessing the antiviral potency of IMU-838 and DHODH-directed back-up drugs in cultured cell-based infection models. By the use of SARS-CoV-2-specific immunofluorescence, Western blot, in-cell ELISA, viral yield reduction and RT-qPCR methods, we demonstrated the following: (i) IMU-838 and back-ups show anti-SARS-CoV-2 activity at several levels of viral replication, i.e., protein production, double-strand RNA synthesis, and release of infectious virus; (ii) antiviral efficacy in Vero cells was demonstrated in a micromolar range (IMU-838 half-maximal effective concentration, EC50, of 7.6 ± 5.8 µM); (iii) anti-SARS-CoV-2 activity was distinct from cytotoxic effects (half-cytotoxic concentration, CC50, >100 µM); (iv) the drug in vitro potency was confirmed using several Vero lineages and human cells; (v) combination with remdesivir showed enhanced anti-SARS-CoV-2 activity; (vi) vidofludimus, the active determinant of IMU-838, exerted a broad-spectrum activity against a selection of major human pathogenic viruses. These findings strongly suggest that developmental DHODH inhibitors represent promising candidates for use as anti-SARS-CoV-2 therapeutics.
Friedrich Hahn; Christina Wangen; Sigrun Häge; Antonia Sophia Peter; Gerhard Dobler; Brett Hurst; Justin Julander; Jonas Fuchs; Zsolt Ruzsics; Klaus Überla; Hans-Martin Jäck; Roger Ptak; Andreas Muehler; Manfred Gröppel; Daniel Vitt; Evelyn Peelen; Hella Kohlhof; Manfred Marschall. IMU-838, a Developmental DHODH Inhibitor in Phase II for Autoimmune Disease, Shows Anti-SARS-CoV-2 and Broad-Spectrum Antiviral Efficacy In Vitro. Viruses 2020, 12, 1394 .
AMA StyleFriedrich Hahn, Christina Wangen, Sigrun Häge, Antonia Sophia Peter, Gerhard Dobler, Brett Hurst, Justin Julander, Jonas Fuchs, Zsolt Ruzsics, Klaus Überla, Hans-Martin Jäck, Roger Ptak, Andreas Muehler, Manfred Gröppel, Daniel Vitt, Evelyn Peelen, Hella Kohlhof, Manfred Marschall. IMU-838, a Developmental DHODH Inhibitor in Phase II for Autoimmune Disease, Shows Anti-SARS-CoV-2 and Broad-Spectrum Antiviral Efficacy In Vitro. Viruses. 2020; 12 (12):1394.
Chicago/Turabian StyleFriedrich Hahn; Christina Wangen; Sigrun Häge; Antonia Sophia Peter; Gerhard Dobler; Brett Hurst; Justin Julander; Jonas Fuchs; Zsolt Ruzsics; Klaus Überla; Hans-Martin Jäck; Roger Ptak; Andreas Muehler; Manfred Gröppel; Daniel Vitt; Evelyn Peelen; Hella Kohlhof; Manfred Marschall. 2020. "IMU-838, a Developmental DHODH Inhibitor in Phase II for Autoimmune Disease, Shows Anti-SARS-CoV-2 and Broad-Spectrum Antiviral Efficacy In Vitro." Viruses 12, no. 12: 1394.
The display of native-like human immunodeficiency virus type 1 envelope (HIV-1 Env) trimers on liposomes has gained wide attention over the last few years. Currently, available methods have enabled the preparation of Env-liposome conjugates of unprecedented quality. However, these protocols require the Env trimer to be tagged and/or to carry a specific functional group. For this reason, we have investigated N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide/N-Hydroxysulfosuccinimide (EDC/Sulfo-NHS) chemistry for its potential to covalently conjugate tag-free, non-functionalized native-like Env trimers onto the surface of carboxyl-functionalized liposomes. The preservation of the liposome’s physical integrity and the immunogen’s conformation required a fine-tuned two-step approach based on the controlled use of β-mercaptoethanol. The display of Env trimers was strictly limited to activated liposomes of positive charge, i.e., liposomes with a positive zeta potential that carry amine-reactive Sulfo-NHS esters on their surface. In agreement with that, conjugation was found to be highly ionic strength- and pH-dependent. Overall, we have identified electrostatic pre-concentration (i.e., close proximity between negatively charged Env trimers and positively charged liposomes established through electrostatic attraction) to be crucial for conjugation reactions to proceed. The present study highlights the requirements and limitations of potentially scalable EDC/Sulfo-NHS-based approaches and represents a solid basis for further research into the controlled conjugation of tag-free, non-functionalized native-like Env trimers on the surface of liposomes, and other nanoparticles.
Ehsan Suleiman; Julia Mayer; Elisabeth Lehner; Bianca Kohlhauser; Alexandra Katholnig; Mirjam Batzoni; Dominik Damm; Vladimir Temchura; Andreas Wagner; Klaus Überla; Karola Vorauer-Uhl. Conjugation of Native-Like HIV-1 Envelope Trimers onto Liposomes Using EDC/Sulfo-NHS Chemistry: Requirements and Limitations. Pharmaceutics 2020, 12, 979 .
AMA StyleEhsan Suleiman, Julia Mayer, Elisabeth Lehner, Bianca Kohlhauser, Alexandra Katholnig, Mirjam Batzoni, Dominik Damm, Vladimir Temchura, Andreas Wagner, Klaus Überla, Karola Vorauer-Uhl. Conjugation of Native-Like HIV-1 Envelope Trimers onto Liposomes Using EDC/Sulfo-NHS Chemistry: Requirements and Limitations. Pharmaceutics. 2020; 12 (10):979.
Chicago/Turabian StyleEhsan Suleiman; Julia Mayer; Elisabeth Lehner; Bianca Kohlhauser; Alexandra Katholnig; Mirjam Batzoni; Dominik Damm; Vladimir Temchura; Andreas Wagner; Klaus Überla; Karola Vorauer-Uhl. 2020. "Conjugation of Native-Like HIV-1 Envelope Trimers onto Liposomes Using EDC/Sulfo-NHS Chemistry: Requirements and Limitations." Pharmaceutics 12, no. 10: 979.
The induction of a potent and long-lasting, broadly neutralizing antibody response is one of the most promising approaches in HIV-1 vaccination. Recently, we demonstrated that Gag-specific T helper cells induced by DNA priming can enhance and modulate the HIV Env-specific B cell response upon virus-like particle (VLP) boost by intrastructural help (ISH). In order to minimize the induction of potentially harmful HIV specific TH cells, we explored the possibility to harness the heterologous TH cells induced by a recombinant tuberculosis subunit vaccine H1, which contains a fusion protein of Ag85B and ESAT-6 antigens in combination with the liposomal adjuvant CAF01. To provide ISH, immunodominant MHC-II restricted peptides from the H1 vaccine were genetically incorporated into the HIV 1 Gag protein and used for HIV VLP production. ISH effects on Env-specific antibody levels and B cell differentiation were analyzed in mice primed against H1 and boosted with VLPs. In contrast to non-primed mice, a significant increase of Env-specific IgG levels for up to 26 weeks after the last immunization was observed. This increase was largely caused by elevated IgG2b and IgG2c levels in mice that received H1 priming. Additionally, ISH enhanced the frequency of Env-specific long-lived plasma cells in the bone marrow. In this study, we were able to demonstrate that a heterologous prime-boost regimen consisting of the H1 tuberculosis subunit vaccine and T helper epitope modified HIV-1 VLPs resulted in enhanced HIV Env antibody and B cell responses, mediated by intrastructural help.
Stephan Klessing; Vladimir Temchura; Pierre Tannig; Antonia Sophia Peter; Dennis Christensen; Roland Lang; Klaus Überla. CD4+ T Cells Induced by Tuberculosis Subunit Vaccine H1 Can Improve the HIV-1 Env Humoral Response by Intrastructural Help. Vaccines 2020, 8, 604 .
AMA StyleStephan Klessing, Vladimir Temchura, Pierre Tannig, Antonia Sophia Peter, Dennis Christensen, Roland Lang, Klaus Überla. CD4+ T Cells Induced by Tuberculosis Subunit Vaccine H1 Can Improve the HIV-1 Env Humoral Response by Intrastructural Help. Vaccines. 2020; 8 (4):604.
Chicago/Turabian StyleStephan Klessing; Vladimir Temchura; Pierre Tannig; Antonia Sophia Peter; Dennis Christensen; Roland Lang; Klaus Überla. 2020. "CD4+ T Cells Induced by Tuberculosis Subunit Vaccine H1 Can Improve the HIV-1 Env Humoral Response by Intrastructural Help." Vaccines 8, no. 4: 604.
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 HIV-1 Rev protein is a nuclear export factor for unspliced and incompletely-spliced HIV-1 RNAs. Without Rev, these intron-retaining RNAs are trapped in the nucleus. A genome-wide screen identified nine proteins of the spliceosome which all enhanced expression from the HIV-1 unspliced RNA after CRISPR/Cas knock-down. Depletion of DHX38, WDR70 and four proteins of the Prp19-associated complex (ISY1, BUD31, XAB2, CRNKL1) resulted in a more than 20-fold enhancement of unspliced HIV-1 RNA levels in the cytoplasm. Targeting of CRNKL1, DHX38, and BUD31 affected nuclear export efficiencies of the HIV-1 unspliced RNA to a much larger extent than splicing. Transcriptomic analyses further revealed that CRNKL1 also suppresses cytoplasmic levels of cellular mRNAs with selectively retained introns. Thus, CRNKL1 dependent nuclear retention seems to be a novel mechanism for the regulation of cytoplasmic levels of intron-retaining cellular mRNAs that is harnessed by HIV-1 to direct its complex splicing pattern.
Han Xiao; Emanuel Wyler; Miha Milek; Bastian Grewe; Philipp Kirchner; Arif Ekici; Ana Beatriz Oliveira Villela Silva; Doris Jungnickl; Markus Landthaler; Armin Ensser; Klaus Überla. CRNKL1 is a highly selective regulator of intron-retaining HIV-1 and cellular mRNAs. 2020, 1 .
AMA StyleHan Xiao, Emanuel Wyler, Miha Milek, Bastian Grewe, Philipp Kirchner, Arif Ekici, Ana Beatriz Oliveira Villela Silva, Doris Jungnickl, Markus Landthaler, Armin Ensser, Klaus Überla. CRNKL1 is a highly selective regulator of intron-retaining HIV-1 and cellular mRNAs. . 2020; ():1.
Chicago/Turabian StyleHan Xiao; Emanuel Wyler; Miha Milek; Bastian Grewe; Philipp Kirchner; Arif Ekici; Ana Beatriz Oliveira Villela Silva; Doris Jungnickl; Markus Landthaler; Armin Ensser; Klaus Überla. 2020. "CRNKL1 is a highly selective regulator of intron-retaining HIV-1 and cellular mRNAs." , no. : 1.
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.
Since the first use of liposomes as carriers for antigens, much work has been done to elucidate the mechanisms involved in the encapsulation of vaccine-relevant biomolecules. However, only a few studies have specifically investigated the encapsulation of hydrophilic, non-conformational peptide epitopes. We performed comprehensive and systematic screening studies, in order to identify conditions that favor the electrostatic interaction of such peptides with lipid membranes. Moreover, we have explored bi-terminal sequence extension as an approach to modify the isoelectric point of peptides, in order to modulate their membrane binding behavior and eventually shift/expand the working range under which they can be efficiently encapsulated in an electrostatically driven manner. The findings of our membrane interaction studies were then applied to preparing peptide-loaded liposomes. Our results show that the magnitude of membrane binding observed in our exploratory in situ setup translates to corresponding levels of encapsulation efficiency in both of the two most commonly employed methods for the preparation of liposomes, i.e., thin-film hydration and microfluidic mixing. We believe that the methods and findings described in the present studies will be of use to a wide audience and can be applied to address the ongoing relevant issue of the efficient encapsulation of hydrophilic biomolecules.
Ehsan Suleiman; Dominik Damm; Mirjam Batzoni; Vladimir Temchura; Andreas Wagner; Klaus Überla; Karola Vorauer-Uhl. Electrostatically Driven Encapsulation of Hydrophilic, Non-Conformational Peptide Epitopes into Liposomes. Pharmaceutics 2019, 11, 619 .
AMA StyleEhsan Suleiman, Dominik Damm, Mirjam Batzoni, Vladimir Temchura, Andreas Wagner, Klaus Überla, Karola Vorauer-Uhl. Electrostatically Driven Encapsulation of Hydrophilic, Non-Conformational Peptide Epitopes into Liposomes. Pharmaceutics. 2019; 11 (11):619.
Chicago/Turabian StyleEhsan Suleiman; Dominik Damm; Mirjam Batzoni; Vladimir Temchura; Andreas Wagner; Klaus Überla; Karola Vorauer-Uhl. 2019. "Electrostatically Driven Encapsulation of Hydrophilic, Non-Conformational Peptide Epitopes into Liposomes." Pharmaceutics 11, no. 11: 619.
Incorporation of immunodominant T-helper epitopes of licensed vaccines into virus-like particles (VLP) allows to harness T-helper cells induced by the licensed vaccines to provide intrastructural help (ISH) for B-cell responses against the surface proteins of the VLPs. To explore whether ISH could also improve antibody responses to calcium phosphate (CaP) nanoparticle vaccines we loaded the nanoparticle core with a universal T-helper epitope of Tetanus toxoid (p30) and functionalized the surface of CaP nanoparticles with stabilized trimers of the HIV-1 envelope (Env) resulting in Env-CaP-p30 nanoparticles. In contrast to soluble Env trimers, Env containing CaP nanoparticles induced activation of naïve Env-specific B-cells in vitro. Mice previously vaccinated against Tetanus raised stronger humoral immune responses against Env after immunization with Env-CaP-p30 than mice not vaccinated against Tetanus. The enhancing effect of ISH on anti-Env antibody levels was not attended with increased Env-specific IFN-γ CD4 T-cell responses that otherwise may potentially influence the susceptibility to HIV-1 infection. Thus, CaP nanoparticles functionalized with stabilized HIV-1 Env trimers and heterologous T-helper epitopes are able to recruit heterologous T-helper cells induced by a licensed vaccine and improve anti-Env antibody responses by intrastructural help.
Dominik Damm; Leonardo Rojas-Sánchez; Hannah Theobald; Viktoriya Sokolova; Richard T. Wyatt; Klaus Überla; Matthias Epple; Vladimir Temchura. Calcium Phosphate Nanoparticle-Based Vaccines as a Platform for Improvement of HIV-1 Env Antibody Responses by Intrastructural Help. Nanomaterials 2019, 9, 1389 .
AMA StyleDominik Damm, Leonardo Rojas-Sánchez, Hannah Theobald, Viktoriya Sokolova, Richard T. Wyatt, Klaus Überla, Matthias Epple, Vladimir Temchura. Calcium Phosphate Nanoparticle-Based Vaccines as a Platform for Improvement of HIV-1 Env Antibody Responses by Intrastructural Help. Nanomaterials. 2019; 9 (10):1389.
Chicago/Turabian StyleDominik Damm; Leonardo Rojas-Sánchez; Hannah Theobald; Viktoriya Sokolova; Richard T. Wyatt; Klaus Überla; Matthias Epple; Vladimir Temchura. 2019. "Calcium Phosphate Nanoparticle-Based Vaccines as a Platform for Improvement of HIV-1 Env Antibody Responses by Intrastructural Help." Nanomaterials 9, no. 10: 1389.
To identify infants with congenital cytomegalovirus (cCMV) saliva polymerase chain reaction (PCR) is an ideal screening method. However, there are only few data on the influence of pre-analytic factors on the analytical sensitivity of the CMV PCR. This study aimed to evaluate the performance of different swabbing materials, transport time and initial virus concentration regarding to the efficacy of recovery of CMV-DNA. Two CMV suspensions containing a high or low concentration of the laboratory strain AD 169 were prepared as test samples. Sampling was simulated by immersion of different swabs in these CMV suspensions and storing the swabs dry or in specified transport media. Transport conditions were modelled by storing the samples for defined time periods prior to DNA extraction and quantitative PCR analyses. Parallel analyses in two different laboratories allowed determination of lab to lab consistency. The duration of storage under the conditions analysed did not have a major effect on the recovery efficiency for the swabbing materials tested. With exception of flocked dry swabs, all tested swabbing materials demonstrated good recovery of CMV DNA. The flocked swab/eNAT system showed the best overall performance. All tested swabbing materials (with exception of the flocked dry swabs) seem to be well suited for recovery of CMV DNA and appropriate for use for the diagnosis of cCMV infection in symptomatic cases and in general cCMV screening programs of newborns.
N. Kohmer; A. Nagel; A. Berger; M. Enders; K. Hamprecht; K. Korn; M. Kortenbusch; K. Überla; H.F. Rabenau. Laboratory diagnosis of congenital CMV infection in newborns: Impact of pre-analytic factors. Journal of Clinical Virology 2019, 115, 32 -36.
AMA StyleN. Kohmer, A. Nagel, A. Berger, M. Enders, K. Hamprecht, K. Korn, M. Kortenbusch, K. Überla, H.F. Rabenau. Laboratory diagnosis of congenital CMV infection in newborns: Impact of pre-analytic factors. Journal of Clinical Virology. 2019; 115 ():32-36.
Chicago/Turabian StyleN. Kohmer; A. Nagel; A. Berger; M. Enders; K. Hamprecht; K. Korn; M. Kortenbusch; K. Überla; H.F. Rabenau. 2019. "Laboratory diagnosis of congenital CMV infection in newborns: Impact of pre-analytic factors." Journal of Clinical Virology 115, no. : 32-36.
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.
Klaus Überla. Die Zoster-Lebendimpfung wird nicht als Standardimpfung empfohlen Attenuated zoster vaccine is not recommended as a standard vaccine. Bundesgesundheitsblatt - Gesundheitsforschung - Gesundheitsschutz 2017, 60, 1065 -1066.
AMA StyleKlaus Überla. Die Zoster-Lebendimpfung wird nicht als Standardimpfung empfohlen Attenuated zoster vaccine is not recommended as a standard vaccine. Bundesgesundheitsblatt - Gesundheitsforschung - Gesundheitsschutz. 2017; 60 (10):1065-1066.
Chicago/Turabian StyleKlaus Überla. 2017. "Die Zoster-Lebendimpfung wird nicht als Standardimpfung empfohlen Attenuated zoster vaccine is not recommended as a standard vaccine." Bundesgesundheitsblatt - Gesundheitsforschung - Gesundheitsschutz 60, no. 10: 1065-1066.
The PAMONO-sensor (plasmon assisted microscopy of nano-objects) demonstrated an ability to detect and quantify individual viruses and virus-like particles. However, another group of biological vesicles—microvesicles (100–1000 nm)—also attracts growing interest as biomarkers of different pathologies and needs development of novel techniques for characterization. This work shows the applicability of a PAMONO-sensor for selective detection of microvesicles in aquatic samples. The sensor permits comparison of relative concentrations of microvesicles between samples. We also study a possibility of repeated use of a sensor chip after elution of the microvesicle capturing layer. Moreover, we improve the detection features of the PAMONO-sensor. The detection process utilizes novel machine learning techniques on the sensor image data to estimate particle size distributions of nano-particles in polydisperse samples. Altogether, our findings expand analytical features and the application field of the PAMONO-sensor. They can also serve for a maturation of diagnostic tools based on the PAMONO-sensor platform.
Victoria Shpacovitch; Irina Sidorenko; Jan Eric Lenssen; Vladimir Temchura; Frank Weichert; Heinrich Müller; Klaus Überla; Alexander Zybin; Alexander Schramm; Roland Hergenröder. Application of the PAMONO-Sensor for Quantification of Microvesicles and Determination of Nano-Particle Size Distribution. Sensors 2017, 17, 244 .
AMA StyleVictoria Shpacovitch, Irina Sidorenko, Jan Eric Lenssen, Vladimir Temchura, Frank Weichert, Heinrich Müller, Klaus Überla, Alexander Zybin, Alexander Schramm, Roland Hergenröder. Application of the PAMONO-Sensor for Quantification of Microvesicles and Determination of Nano-Particle Size Distribution. Sensors. 2017; 17 (2):244.
Chicago/Turabian StyleVictoria Shpacovitch; Irina Sidorenko; Jan Eric Lenssen; Vladimir Temchura; Frank Weichert; Heinrich Müller; Klaus Überla; Alexander Zybin; Alexander Schramm; Roland Hergenröder. 2017. "Application of the PAMONO-Sensor for Quantification of Microvesicles and Determination of Nano-Particle Size Distribution." Sensors 17, no. 2: 244.