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1. Correction for Kibler et al., “Replication-Competent NYVAC-KC Yields Improved Immunogenicity to HIV-1 Antigens in Rhesus Macaques Compared to Nonreplicating NYVAC”

Author

Kibler, Karen V, Asbach, Benedikt, Perdiguero, Beatriz, García-Arriaza, Juan, Yates, Nicole L, Parks, Robert, Stanfield-Oakley, Sherry, Ferrari, Guido, Montefiori, David C, Tomaras, Georgia D, Roederer, Mario, Foulds, Kathryn E, Forthal, Donald N, Seaman, Michael S, Self, Steve, Gottardo, Raphael, Phogat, Sanjay, Tartaglia, James, Barnett, Susan, Cristillo, Anthony D, Weiss, Deborah, Galmin, Lindsey, Ding, Song, Heeney, Jonathan L, Esteban, Mariano, Wagner, Ralf, Pantaleo, Giuseppe, and Jacobs, Bertram L

Subjects
Medical Microbiology, Biomedical and Clinical Sciences, Immunology, Good Health and Well Being, Biological Sciences, Agricultural and Veterinary Sciences, Medical and Health Sciences, Virology, Agricultural, veterinary and food sciences, Biological sciences, Biomedical and clinical sciences
Abstract

Volume 93, no. 3, e01513-18, 2019, https://doi.org/10.1128/JVI.01513-18. The article was originally published on 17 January 2019 with a standard copyright line (“© 2019 American Society for Microbiology. All Rights Reserved.”). We elected to pay for open access for the article after publication, necessitating replacement of the original copyright line with “© 2019 Kibler et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.” This change was made to the online version of the article on 21 August 2019.

Published
2019

2. A serum-stable RNA aptamer specific for SARS-CoV-2 neutralizes viral entry

Author

Valero, Julián, Civit, Laia, Dupont, Daniel M., Selnihhin, Denis, Reinert, Line S., Idorn, Manja, Israels, Brett A., Bednarz, Aleksandra M., Bus, Claus, Asbach, Benedikt, Peterhoff, David, Pedersen, Finn S., Birkedal, Victoria, Wagner, Ralf, Paludan, Søren R., and Kjems, Jørgen

Published
2021

5. Replication-Competent NYVAC-KC Yields Improved Immunogenicity to HIV-1 Antigens in Rhesus Macaques Compared to Nonreplicating NYVAC.

Author

Kibler, Karen V, Asbach, Benedikt, Perdiguero, Beatriz, García-Arriaza, Juan, Yates, Nicole L, Parks, Robert, Stanfield-Oakley, Sherry, Ferrari, Guido, Montefiori, David C, Tomaras, Georgia D, Roederer, Mario, Foulds, Kathryn E, Forthal, Donald N, Seaman, Michael S, Self, Steve, Gottardo, Raphael, Phogat, Sanjay, Tartaglia, James, Barnett, Susan, Cristillo, Anthony D, Weiss, Deborah, Galmin, Lindsey, Ding, Song, Heeney, Jonathan L, Esteban, Mariano, Wagner, Ralf, Pantaleo, Giuseppe, and Jacobs, Bertram L

Subjects
Medical Microbiology, Biomedical and Clinical Sciences, Immunology, Vaccine Related (AIDS), Biotechnology, Prevention, Immunization, HIV/AIDS, Vaccine Related, Infectious Diseases, Prevention of disease and conditions, and promotion of well-being, 3.4 Vaccines, Infection, Good Health and Well Being, Animals, Antibodies, Neutralizing, CD4-Positive T-Lymphocytes, HIV Antibodies, HIV Antigens, HIV Infections, HIV-1, Humans, Macaca mulatta, Male, Vaccination, Vaccinia virus, Viral Vaccines, Virus Replication, env Gene Products, Human Immunodeficiency Virus, Gag-Pol-Nef, HIV, NYVAC, NYVAC-KC, T cell response, antibody responses, gp140, nonhuman primates, vaccines, Biological Sciences, Agricultural and Veterinary Sciences, Medical and Health Sciences, Virology, Agricultural, veterinary and food sciences, Biological sciences, Biomedical and clinical sciences
Abstract

As part of the continuing effort to develop an effective HIV vaccine, we generated a poxviral vaccine vector (previously described) designed to improve on the results of the RV144 phase III clinical trial. The construct, NYVAC-KC, is a replication-competent, attenuated recombinant of the vaccinia virus strain NYVAC. NYVAC is a vector that has been used in many previous clinical studies but is replication deficient. Here, we report a side-by-side comparison of replication-restricted NYVAC and replication-competent NYVAC-KC in a nonhuman primate study, which utilized a prime-boost regimen similar to that of RV144. NYVAC-C and NYVAC-C-KC express the HIV-1 antigens gp140, and Gag/Gag-Pol-Nef-derived virus-like particles (VLPs) from clade C and were used as the prime, with recombinant virus plus envelope protein used as the boost. In nearly every T and B cell immune assay against HIV-1, including neutralization and antibody binding, NYVAC-C-KC induced a greater immune response than NYVAC-C, indicating that replication competence in a poxvirus may improve upon the modestly successful regimen used in the RV144 clinical trial.IMPORTANCE Though the RV144 phase III clinical trial showed promise that an effective vaccine against HIV-1 is possible, a successful vaccine will require improvement over the vaccine candidate (ALVAC) used in the RV144 study. With that goal in mind, we have tested in nonhuman primates an attenuated but replication-competent vector, NYVAC-KC, in direct comparison to its parental vector, NYVAC, which is replication restricted in human cells, similar to the ALVAC vector used in RV144. We have utilized a prime-boost regimen for administration of the vaccine candidate that is similar to the one used in the RV144 study. The results of this study indicate that a replication-competent poxvirus vector may improve upon the effectiveness of the RV144 clinical trial vaccine candidate.

Published
2019

6. Priming with a Potent HIV-1 DNA Vaccine Frames the Quality of Immune Responses prior to a Poxvirus and Protein Boost.

Author

Asbach, Benedikt, Kibler, Karen V, Köstler, Josef, Perdiguero, Beatriz, Yates, Nicole L, Stanfield-Oakley, Sherry, Tomaras, Georgia D, Kao, Shing-Fen, Foulds, Kathryn E, Roederer, Mario, Seaman, Michael S, Montefiori, David C, Parks, Robert, Ferrari, Guido, Forthal, Donald N, Phogat, Sanjay, Tartaglia, James, Barnett, Susan W, Self, Steven G, Gottardo, Raphael, Cristillo, Anthony D, Weiss, Deborah E, Galmin, Lindsey, Ding, Song, Heeney, Jonathan L, Esteban, Mariano, Jacobs, Bertram L, Pantaleo, Giuseppe, and Wagner, Ralf

Subjects
Medical Microbiology, Biomedical and Clinical Sciences, Immunology, Prevention, Vaccine Related, HIV/AIDS, Biotechnology, Immunization, Rare Diseases, Vaccine Related (AIDS), Prevention of disease and conditions, and promotion of well-being, 3.4 Vaccines, Infection, Good Health and Well Being, Animals, Antibodies, Neutralizing, HIV Antibodies, HIV Antigens, HIV Infections, HIV-1, Humans, Macaca mulatta, Male, Poxviridae, T-Lymphocytes, Vaccination, Vaccines, DNA, Vaccinia virus, Viral Vaccines, Virus Replication, DNA vaccine, Gag-Pol-Nef, NYVAC, NYVAC-KC, T cell responses, antibody responses, gp140, human immunodeficiency virus, nonhuman primates, vaccine, Biological Sciences, Agricultural and Veterinary Sciences, Medical and Health Sciences, Virology, Agricultural, veterinary and food sciences, Biological sciences, Biomedical and clinical sciences
Abstract

The use of heterologous immunization regimens and improved vector systems has led to increases in immunogenicity of HIV-1 vaccine candidates in nonhuman primates. In order to resolve interrelations between different delivery modalities, three different poxvirus boost regimens were compared. Three groups of rhesus macaques were each primed with the same DNA vaccine encoding Gag, Pol, Nef, and gp140. The groups were then boosted with either the vaccinia virus strain NYVAC or a variant with improved replication competence in human cells, termed NYVAC-KC. The latter was administered either by scarification or intramuscularly. Finally, macaques were boosted with adjuvanted gp120 protein to enhance humoral responses. The regimen elicited very potent CD4+ and CD8+ T cell responses in a well-balanced manner, peaking 2 weeks after the boost. T cells were broadly reactive and polyfunctional. All animals exhibited antigen-specific humoral responses already after the poxvirus boost, which further increased following protein administration. Polyclonal reactivity of IgG antibodies was highest against HIV-1 clade C Env proteins, with considerable cross-reactivity to other clades. Substantial effector functional activities (antibody-dependent cell-mediated cytotoxicity and antibody-dependent cell-mediated virus inhibition) were observed in serum obtained after the last protein boost. Notably, major differences between the groups were absent, indicating that the potent priming induced by the DNA vaccine initially framed the immune responses in such a way that the subsequent boosts with NYVAC and protein led only to an increase in the response magnitudes without skewing the quality. This study highlights the importance of selecting the best combination of vector systems in heterologous prime-boost vaccination regimens.IMPORTANCE The evaluation of HIV vaccine efficacy trials indicates that protection would most likely correlate with a polyfunctional immune response involving several effector functions from all arms of the immune system. Heterologous prime-boost regimens have been shown to elicit vigorous T cell and antibody responses in nonhuman primates that, however, qualitatively and quantitatively differ depending on the respective vector systems used. The present study evaluated a DNA prime and poxvirus and protein boost regimen and compared how two poxvirus vectors with various degrees of replication capacity and two different delivery modalities-conventional intramuscular delivery and percutaneous delivery by scarification-impact several immune effectors. It was found that despite the different poxvirus boosts, the overall immune responses in the three groups were similar, suggesting the potent DNA priming as the major determining factor of immune responses. These findings emphasize the importance of selecting optimal priming agents in heterologous prime-boost vaccination settings.

Published
2019

7. Transgene expression knock-down in recombinant Modified Vaccinia virus Ankara vectors improves genetic stability and sustained transgene maintenance across multiple passages

Author

Neckermann, Patrick, primary, Mohr, Madlen, additional, Billmeier, Martina, additional, Karlas, Alexander, additional, Boilesen, Ditte R., additional, Thirion, Christian, additional, Holst, Peter J., additional, Jordan, Ingo, additional, Sandig, Volker, additional, Asbach, Benedikt, additional, and Wagner, Ralf, additional

Published
2024
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8. Potential To Streamline Heterologous DNA Prime and NYVAC/Protein Boost HIV Vaccine Regimens in Rhesus Macaques by Employing Improved Antigens

Author

Asbach, Benedikt, Kliche, Alexander, Köstler, Josef, Perdiguero, Beatriz, Esteban, Mariano, Jacobs, Bertram L, Montefiori, David C, LaBranche, Celia C, Yates, Nicole L, Tomaras, Georgia D, Ferrari, Guido, Foulds, Kathryn E, Roederer, Mario, Landucci, Gary, Forthal, Donald N, Seaman, Michael S, Hawkins, Natalie, Self, Steven G, Sato, Alicia, Gottardo, Raphael, Phogat, Sanjay, Tartaglia, James, Barnett, Susan W, Burke, Brian, Cristillo, Anthony D, Weiss, Deborah E, Francis, Jesse, Galmin, Lindsey, Ding, Song, Heeney, Jonathan L, Pantaleo, Giuseppe, Wagner, Ralf, and Silvestri, G.

Published
2016

9. Preferential Expansion of HPV16 E1-Specific T Cells from Healthy Donors’ PBMCs after Ex Vivo Immunization with an E1E2E6E7 Fusion Antigen

Author

Daradoumis, Joana, primary, Müller, Mikkel Dons, additional, Neckermann, Patrick, additional, Asbach, Benedikt, additional, Schrödel, Silke, additional, Thirion, Christian, additional, Wagner, Ralf, additional, thor Straten, Per, additional, Holst, Peter Johannes, additional, and Boilesen, Ditte, additional

Published
2023
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10. Potency and durability of T and B cell immune responses after homologous and heterologous vector delivery of a trimer-stabilized, membrane-displayed HIV-1 clade ConC Env protein

Author

Perdiguero, Beatriz, primary, Hauser, Alexandra, additional, Gómez, Carmen Elena, additional, Peterhoff, David, additional, Sideris, Elefthéria, additional, Sorzano, Carlos Óscar S., additional, Wilmschen, Sarah, additional, Schaber, Marion, additional, Stengel, Laura, additional, Asbach, Benedikt, additional, Ding, Song, additional, Von Laer, Dorothee, additional, Levy, Yves, additional, Pantaleo, Giuseppe, additional, Kimpel, Janine, additional, Esteban, Mariano, additional, and Wagner, Ralf, additional

Published
2023
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11. Transgene expression knockdown in recombinant Modified Vaccinia virus Ankara vectors improves genetic stability and sustained transgene maintenance across multiple passages.

Author

Neckermann, Patrick, Mohr, Madlen, Billmeier, Martina, Karlas, Alexander, Boilesen, Ditte R., Thirion, Christian, Holst, Peter J., Jordan, Ingo, Sandig, Volker, Asbach, Benedikt, and Wagner, Ralf

Subjects
TRANSGENE expression, VACCINIA, SUPPRESSOR cells, KRA, NUCLEOTIDE sequencing, GENETIC transformation, GENETIC vectors
Abstract

Modified vaccinia virus Ankara is a versatile vaccine vector, well suited for transgene delivery, with an excellent safety profile. However, certain transgenes render recombinant MVA (rMVA) genetically unstable, leading to the accumulation of mutated rMVA with impaired transgene expression. This represents a major challenge for upscaling and manufacturing of rMVA vaccines. To prevent transgene-mediated negative selection, the continuous avian cell line AGE1.CR pIX (CR pIX) was modified to suppress transgene expression during rMVA generation and amplification. This was achieved by constitutively expressing a tetracycline repressor (TetR) together with a rat-derived shRNA in engineered CR pIX PRO suppressor cells targeting an operator element (tetO) and 3' untranslated sequence motif on a chimeric poxviral promoter and the transgene mRNA, respectively. This cell line was instrumental in generating two rMVA (isolate CR19) expressing a Macaca fascicularis papillomavirus type 3 (MfPV3) E1E2E6E7 artificially-fused polyprotein following recombinationmediated integration of the coding sequences into the DelIII (CR19 M-DelIII) or TK locus (CR19 M-TK), respectively. Characterization of rMVA on parental CR pIX or engineered CR pIX PRO suppressor cells revealed enhanced replication kinetics, higher virus titers and a focus morphology equaling wild-type MVA, when transgene expression was suppressed. Serially passaging both rMVA ten times on parental CR pIX cells and tracking E1E2E6E7 expression by flow cytometry revealed a rapid loss of transgene product after only few passages. PCR analysis and next-generation sequencing demonstrated that rMVA accumulated mutations within the E1E2E6E7 open reading frame (CR19 M-TK) or deletions of the whole transgene cassette (CR19 M-DelIII). In contrast, CR pIX PRO suppressor cells preserved robust transgene expression for up to 10 passages, however, rMVAs were more stable when E1E2E6E7 was integrated into the TK as compared to the DelIII locus. In conclusion, sustained knock-down of transgene expression in CR pIX PRO suppressor cells facilitates the generation, propagation and large-scale manufacturing of rMVA with transgenes hampering viral replication. [ABSTRACT FROM AUTHOR]

Published
2024
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12. A computationally designed antigen eliciting broad humoral responses against SARS-CoV-2 and related sarbecoviruses

Author

Vishwanath, Sneha, primary, Carnell, George William, additional, Ferrari, Matteo, additional, Asbach, Benedikt, additional, Billmeier, Martina, additional, George, Charlotte, additional, Sans, Maria Suau, additional, Nadesalingam, Angalee, additional, Huang, Chloe Qingzhou, additional, Paloniemi, Minna, additional, Stewart, Hazel, additional, Chan, Andrew, additional, Wells, David Arthur, additional, Neckermann, Patrick, additional, Peterhoff, David, additional, Einhauser, Sebastian, additional, Cantoni, Diego, additional, Neto, Martin Mayora, additional, Jordan, Ingo, additional, Sandig, Volker, additional, Tonks, Paul, additional, Temperton, Nigel, additional, Frost, Simon, additional, Sohr, Katharina, additional, Ballesteros, Maria Teresa Lluesma, additional, Arbabi, Farzad, additional, Geiger, Johannes, additional, Dohmen, Christian, additional, Plank, Christian, additional, Kinsley, Rebecca, additional, Wagner, Ralf, additional, and Heeney, Jonathan Luke, additional

Published
2023
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13. Supplementary Data from Efficacy and Synergy with Cisplatin of an Adenovirus Vectored Therapeutic E1E2E6E7 Vaccine against HPV Genome–Positive C3 Cancers in Mice

Author

Boilesen, Ditte Rahbæk, primary, Neckermann, Patrick, primary, Willert, Torsten, primary, Müller, Mikkel Dons, primary, Schrödel, Silke, primary, Pertl, Cordula, primary, Thirion, Christian, primary, Asbach, Benedikt, primary, Wagner, Ralf, primary, and Holst, Peter Johannes, primary

Published
2023
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16. Glycan masking of a non-neutralising epitope enhances neutralising antibodies targeting the RBD of SARS-CoV-2 and its variants

Author

Carnell, George W., primary, Billmeier, Martina, additional, Vishwanath, Sneha, additional, Suau Sans, Maria, additional, Wein, Hannah, additional, George, Charlotte L., additional, Neckermann, Patrick, additional, Del Rosario, Joanne Marie M., additional, Sampson, Alexander T., additional, Einhauser, Sebastian, additional, Aguinam, Ernest T., additional, Ferrari, Matteo, additional, Tonks, Paul, additional, Nadesalingam, Angalee, additional, Schütz, Anja, additional, Huang, Chloe Qingzhou, additional, Wells, David A., additional, Paloniemi, Minna, additional, Jordan, Ingo, additional, Cantoni, Diego, additional, Peterhoff, David, additional, Asbach, Benedikt, additional, Sandig, Volker, additional, Temperton, Nigel, additional, Kinsley, Rebecca, additional, Wagner, Ralf, additional, and Heeney, Jonathan L., additional

Published
2023
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17. DNA origami traps for large viruses

Author

Monferrer, Alba, primary, Kohler, Fabian, additional, Sigl, Christian, additional, Schachtner, Michael, additional, Peterhoff, David, additional, Asbach, Benedikt, additional, Wagner, Ralf, additional, and Dietz, Hendrik, additional

Published
2023
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18. Efficacy and Synergy with Cisplatin of an Adenovirus Vectored Therapeutic E1E2E6E7 Vaccine against HPV Genome–Positive C3 Cancers in Mice

Author

Boilesen, Ditte Rahbæk, primary, Neckermann, Patrick, additional, Willert, Torsten, additional, Müller, Mikkel Dons, additional, Schrödel, Silke, additional, Pertl, Cordula, additional, Thirion, Christian, additional, Asbach, Benedikt, additional, Wagner, Ralf, additional, and Holst, Peter Johannes, additional

Published
2022
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19. Influenza A (N1-N9) and Influenza B (B/Victoria and B/Yamagata) Neuraminidase Pseudotypes as Tools for Pandemic Preparedness and Improved Influenza Vaccine Design

Author

da Costa, Kelly A. S., primary, Del Rosario, Joanne Marie M., additional, Ferrari, Matteo, additional, Vishwanath, Sneha, additional, Asbach, Benedikt, additional, Kinsley, Rebecca, additional, Wagner, Ralf, additional, Heeney, Jonathan L., additional, Carnell, George W., additional, and Temperton, Nigel J., additional

Published
2022
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20. Early and Long-Term HIV-1 Immunogenicity Induced in Macaques by the Combined Administration of DNA, NYVAC and Env Protein-Based Vaccine Candidates: The AUP512 Study

Author

Perdiguero, Beatriz, primary, Asbach, Benedikt, additional, Gómez, Carmen E., additional, Köstler, Josef, additional, Barnett, Susan W., additional, Koutsoukos, Marguerite, additional, Weiss, Deborah E., additional, Cristillo, Anthony D., additional, Foulds, Kathryn E., additional, Roederer, Mario, additional, Montefiori, David C., additional, Yates, Nicole L., additional, Ferrari, Guido, additional, Shen, Xiaoying, additional, Sawant, Sheetal, additional, Tomaras, Georgia D., additional, Sato, Alicia, additional, Fulp, William J., additional, Gottardo, Raphael, additional, Ding, Song, additional, Heeney, Jonathan L., additional, Pantaleo, Giuseppe, additional, Esteban, Mariano, additional, and Wagner, Ralf, additional

Published
2022
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22. Comparative Immunogenicity of COVID-19 Vaccines in a Population-Based Cohort Study with SARS-CoV-2-Infected and Uninfected Participants

Author

Peterhoff, David, Einhauser, Sebastian, Beileke, Stephanie, Niller, Hans-Helmut, Günther, Felix, Schachtner, Michael, Asbach, Benedikt, Steininger, Philipp, Tenbusch, Matthias, Peter, Antonia S., Gessner, Andre, Burkhardt, Ralph, Heid, Iris M., Wagner, Ralf, and Überla, Klaus

Subjects
Pharmacology, ddc:610, Infectious Diseases, Drug Discovery, Immunology, 610 Medizin, Pharmacology (medical), SARS-CoV-2, vaccination, population-based cohort, immunogenicity, neutralizing antibodies, Comirnaty, Spikevax, Vaxzevria, variants of concern VoC
Abstract

To assess vaccine immunogenicity in non-infected and previously infected individuals in a real-world scenario, SARS-CoV-2 antibody responses were determined during follow-up 2 (April 2021) of the population-based Tirschenreuth COVID-19 cohort study comprising 3378 inhabitants of the Tirschenreuth county aged 14 years or older. Seronegative participants vaccinated once with Vaxzevria, Comirnaty, or Spikevax had median neutralizing antibody titers ranging from ID50 = 25 to 75. Individuals with two immunizations with Comirnaty or Spikevax had higher median ID50s (of 253 and 554, respectively). Regression analysis indicated that both increased age and increased time since vaccination independently decreased RBD binding and neutralizing antibody levels. Unvaccinated participants with detectable N-antibodies at baseline (June 2020) revealed a median ID50 of 72 at the April 2021 follow-up. Previously infected participants that received one dose of Vaxzevria or Comirnaty had median ID50 to 929 and 2502, respectively. Individuals with a second dose of Comirnaty given in a three-week interval after the first dose did not have higher median antibody levels than individuals with one dose. Prior infection also primed for high systemic IgA levels in response to one dose of Comirnaty that exceeded IgA levels observed after two doses of Comirnaty in previously uninfected participants. Neutralizing antibody levels targeting the spike protein of Beta and Delta variants were diminished compared to the wild type in vaccinated and infected participants.

Published
2022
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23. Antibodies against viral nucleo-, phospho-, and X protein contribute to serological diagnosis of fatal Borna disease virus 1 infections

Author

Neumann, Bernhard, Angstwurm, Klemens, Linker, Ralf A., Knoll, Gertrud, Eidenschink, Lisa, Rubbenstroth, Dennis, Schlottau, Kore, Beer, Martin, Schreiner, Patrick, Soutschek, Erwin, Böhmer, Merle M., Lampl, Benedikt M.J., Pregler, Matthias, Scheiter, Alexander, Evert, Katja, Zoubaa, Saida, Riemenschneider, Markus J., Asbach, Benedikt, Gessner, André, Niller, Hans Helmut, Schmidt, Barbara, and Bauswein, Markus

Subjects
Borna disease virus 1, encephalitis, encephalopathy, zoonosis, recombinant protein, diagnostics, humoral immune response, antibodies, ELISA, encephalitis, 610 Medizin, General Biochemistry, Genetics and Molecular Biology, Antibodies, Viral Proteins, Report, Chlorocebus aethiops, diagnostics, 570 Biowissenschaften, Biologie, Animals, Humans, Borna disease virus, Vero Cells, Aged, ddc:610, Borna disease virus 1, zoonosis, encephalopathy, Phosphoproteins, humoral immune response, Recombinant Proteins, Nucleoproteins, Borna Disease, ELISA, ddc:570, recombinant protein
Abstract

Summary Borna disease virus 1 (BoDV-1) causes rare but often fatal encephalitis in humans. Late diagnosis prohibits an experimental therapeutic approach. Here, we report a recent case of fatal BoDV-1 infection diagnosed on day 12 after hospitalization by detection of BoDV-1 RNA in the cerebrospinal fluid. In a retrospective analysis, we detect BoDV-1 RNA 1 day after hospital admission when the cell count in the cerebrospinal fluid is still normal. We develop a new ELISA using recombinant BoDV-1 nucleoprotein, phosphoprotein, and accessory protein X to detect seroconversion on day 12. Antibody responses are also shown in seven previously confirmed cases. The individual BoDV-1 antibody profiles show variability, but the usage of three different BoDV-1 antigens results in a more sensitive diagnostic tool. Our findings demonstrate that early detection of BoDV-1 RNA in cerebrospinal fluid and the presence of antibodies against at least two different viral antigens contribute to BoDV-1 diagnosis. Physicians in endemic regions should consider BoDV-1 infection in cases of unclear encephalopathy and initiate appropriate diagnostics at an early stage., Graphical abstract, Highlights • Borna disease virus 1 causes fatal encephalitis upon zoonotic spillover infections • An ELISA system using recombinant BoDV-1 N, X, and P proteins has been established • Antibodies against at least two different BoDV-1 antigens corroborate seroconversion • Early detection of viral RNA and antibodies could contribute to BoDV-1 diagnosis, Neumann et al. provide data on antibody responses to Borna disease virus 1 in eight cases of fatal encephalitis based on a newly developed ELISA using recombinant BoDV-1 nucleoprotein, phosphoprotein, and accessory protein X. The presence of antibodies against at least two different viral antigens contributes to BoDV-1 diagnosis.

Published
2022

24. Comparative Immunogenicity of COVID-19 Vaccines in a Population-Based Cohort Study with SARS-CoV-2-Infected and Uninfected Participants

Author

Peterhoff, David, primary, Einhauser, Sebastian, additional, Beileke, Stephanie, additional, Niller, Hans-Helmut, additional, Günther, Felix, additional, Schachtner, Michael, additional, Asbach, Benedikt, additional, Steininger, Philipp, additional, Tenbusch, Matthias, additional, Peter, Antonia S., additional, Gessner, Andre, additional, Burkhardt, Ralph, additional, Heid, Iris M., additional, Wagner, Ralf, additional, and Überla, Klaus, additional

Published
2022
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25. Gene delivery of a single,structurally engineered Coronavirus vaccine antigen elicits SARS-CoV-2 Omicron and pan-Sarbecovirus neutralisation

Author

Heeney, Jonathan, primary, Vishwanath, Sneha, additional, Carnell, George, additional, Ferrari, Matteo, additional, Asbach, Benedikt, additional, Billmeier, Martina, additional, George, Charlotte, additional, Sans, Maria Suau, additional, Neckermann, Patrick, additional, Peterhoff, David, additional, Cantoni, Diego, additional, Wells, David, additional, Temperton, Nigel, additional, and Frost, Simon, additional

Published
2022
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26. Design and Immunological Validation of Macaca fascicularis Papillomavirus Type 3 Based Vaccine Candidates in Outbred Mice: Basis for Future Testing of a Therapeutic Papillomavirus Vaccine in NHPs

Author

Neckermann, Patrick, primary, Boilesen, Ditte Rahbaek, additional, Willert, Torsten, additional, Pertl, Cordula, additional, Schrödel, Silke, additional, Thirion, Christian, additional, Asbach, Benedikt, additional, Holst, Peter Johannes, additional, and Wagner, Ralf, additional

Published
2021
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27. Influenza A (N1-N9) and Influenza B (B/Vic and B/Yam) Neuraminidase Pseudotypes as Tools for Pandemic Preparedness and Improved Influenza Vaccine design

Author

da Costa, Kelly A.S., primary, Del Rosario, Joanne Marie M., additional, Ferrari, Matteo, additional, Vishwanath, Sneha, additional, Asbach, Benedikt, additional, Kinsley, Rebecca, additional, Wagner, Ralf, additional, Heeney, Jonathan L, additional, Carnell, George W, additional, and Temperton, Nigel James, additional

Published
2021
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28. Coronavirus Pseudotypes for All Circulating Human Coronaviruses for Quantification of Cross-Neutralizing Antibody Responses

Author

Sampson, Alexander Thomas, primary, Heeney, Jonathan, additional, Cantoni, Diego, additional, Ferrari, Matteo, additional, Sans, Maria Suau, additional, George, Charlotte, additional, Di Genova, Cecilia, additional, Mayora Neto, Martin, additional, Einhauser, Sebastian, additional, Asbach, Benedikt, additional, Wagner, Ralf, additional, Baxendale, Helen, additional, Temperton, Nigel, additional, and Carnell, George, additional

Published
2021
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29. SARS-CoV-2 Spike Protein Stabilized in the Closed State Induces Potent Neutralizing Responses

Author

Carnell, George W., primary, Ciazynska, Katarzyna A., additional, Wells, David A., additional, Xiong, Xiaoli, additional, Aguinam, Ernest T., additional, McLaughlin, Stephen H., additional, Mallery, Donna, additional, Ebrahimi, Soraya, additional, Ceron-Gutierrez, Lourdes, additional, Asbach, Benedikt, additional, Einhauser, Sebastian, additional, Wagner, Ralf, additional, James, Leo C., additional, Doffinger, Rainer, additional, Heeney, Jonathan L., additional, and Briggs, John A. G., additional

Published
2021
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30. Exploiting Pan Influenza A and Pan Influenza B Pseudotype Libraries for Efficient Vaccine Antigen Selection

Author

Del Rosario, Joanne Marie M., primary, da Costa, Kelly A. S., additional, Asbach, Benedikt, additional, Ferrara, Francesca, additional, Ferrari, Matteo, additional, Wells, David A., additional, Mann, Gurdip Singh, additional, Ameh, Veronica O., additional, Sabeta, Claude T., additional, Banyard, Ashley C., additional, Kinsley, Rebecca, additional, Scott, Simon D., additional, Wagner, Ralf, additional, Heeney, Jonathan L., additional, Carnell, George W., additional, and Temperton, Nigel J., additional

Published
2021
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31. Coronavirus pseudotypes for all circulating human coronaviruses for quantification of cross-neutralising antibody responses

Author

Sampson, Alex, Heeney, Jl, Ferrari, Matteo, George, Charlotte, Wagner, Ralf, Di Genova, Cecilia, Cantoni, Diego, Mayora-Neto, Martin, Einhauser, Sebastian, Asbach, Benedikt, Baxendale, Helen, Temperton, Nigel, and Carnell, George

Subjects
viruses, virus diseases
Abstract

The novel coronavirus SARS-CoV-2 is the seventh identified human coronavirus. Understanding the extent of pre-existing immunity induced by seropositivity to endemic seasonal coronaviruses and the impact of cross-reactivity on COVID-19 disease progression remains a key research question in immunity to SARS-CoV-2 and the immunopathology of COVID-2019 disease. This paper describes a panel of lentiviral pseudotypes bearing the spike (S) proteins for each of the seven human coronaviruses (HCoVs), generated under similar conditions optimised for high titre production allowing high-throughput investigation of antibody neutralisation breadth. Optimal production conditions and most readily available permissive target cell lines were determined for spike-mediated entry by each HCoV pseudotype – SARS-CoV-1, SARS-CoV-2 and HCoV-NL63 best transduced HEK293T/17 cells transfected with ACE2 and TMPRSS2. HCoV-229E and MERS-CoV preferentially entered Huh7 cells, and CHO cells were most permissive for the seasonal betacoronavirus HCoV-HKU1. Entry of ACE-2 using pseudotypes was enhanced by ACE-2 and TMPRSS2 expression in target cells, whilst TMPRSS2 transfection rendered HEK293T/17 cells permissive for HCoV-HKU1 and HCoV-OC43 entry. Additionally, pseudotype viruses were produced bearing additional coronavirus surface proteins including SARS-CoV-2 Envelope (E) and Membrane (M) proteins and HCoV-OC43/HCoV-HKU1 Haemagglutinin-Esterase (HE) proteins. This panel of lentiviral pseudotypes provides a safe, rapidly quantifiable and high-throughput tool for serological comparison of pan-coronavirus neutralising responses; this can be used to elucidate antibody dynamics against individual coronaviruses and the effects of antibody cross-reactivity on clinical outcome following natural infection or vaccination.

Published
2021
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33. Establishment of pan-Influenza A (H1-H18) and pan-Influenza B (pre-split, Vic/Yam) Pseudotype Libraries for efficient vaccine antigen selection

Author

Del Rosario, Joanne Marie M., primary, da Costa, Kelly A.S., additional, Asbach, Benedikt, additional, Ferrara, Francesca, additional, Ferrari, Matteo, additional, Wells, David A., additional, Mann, Gurdip Singh, additional, Ameh, Veronica O., additional, Sabeta, Claude T., additional, Banyard, Ashley C., additional, Kinsley, Rebecca, additional, Scott, Simon D., additional, Wagner, Ralf, additional, Heeney, Jonathan L., additional, Carnell, George W., additional, and Temperton, Nigel J., additional

Published
2021
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35. 35 Jahre HIV - Ist ein Ende der Pandemie in Sicht?

Author

Asbach, Benedikt, Peterhoff, David, and Wagner, Ralf

Abstract

Eine Diagnose »HIV-positiv« war Anfang der Neunzigerjahre mit einer ungünstigen Prognose, einer Überlebenserwartung von nur wenigen Jahren und einem Sterben unter qualvollsten Umständen verknüpft. Heute, gut 35 Jahre später, ist die Infektion in der entwickelten Welt zwar nicht heilbar, aber doch so gut behandelbar, dass Lebenserwartung und -qualität kaum beeinträchtigt sind. Trotz der unglaublichen Fortschritte bei der Behandlung der Erkrankung und der Möglichkeit, sich durch die Einnahme von Medikamentencocktails kurzfristig vor einer Infektion zu schützen, würden insbesondere die Menschen in Entwicklungs- und Schwellenländern nachhaltig von der Verfügbarkeit eines vor HIVInfektion schützenden Impfstoffes profitieren. Dieser Artikel soll die wesentlichen Meilensteine auf dem Weg zur Begrenzung der globalen HIV-Epidemie zusammenfassen, die Probleme auf dem Weg zu einem effektiven Impfstoff skizzieren und den Beitrag unserer Universität im internationalen Bemühen um einen HIV-Impfstoff aufzeigen., Blick in die Wissenschaft, Bd. 28 Nr. 40 (2019): Blick in die Wissenschaft 40

Published
2019
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36. Heterologous Combination of VSV-GP and NYVAC Vectors Expressing HIV-1 Trimeric gp145 Env as Vaccination Strategy to Induce Balanced B and T Cell Immune Responses

Author

Perdiguero, Beatriz, primary, Gómez, Carmen Elena, additional, García-Arriaza, Juan, additional, Sánchez-Corzo, Cristina, additional, Sorzano, Carlos Óscar S., additional, Wilmschen, Sarah, additional, von Laer, Dorothee, additional, Asbach, Benedikt, additional, Schmalzl, Christina, additional, Peterhoff, David, additional, Ding, Song, additional, Wagner, Ralf, additional, Kimpel, Janine, additional, Levy, Yves, additional, Pantaleo, Giuseppe, additional, and Esteban, Mariano, additional

Published
2019
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38. Additional File 1

Author

Barabas, Sascha, Spindler, Theresa, Kiener, Richard, Tonar, Charlotte, Lugner, Tamara, Batzilla, Julia, Bendfeldt, Hanna, Rascle, Anne, Asbach, Benedikt, Wagner, Ralf, and Deml, Ludwig

Abstract

Assay variability. Intra-assay (Table S1.), inter-assay (Table S2.), inter-operator (Table S3.) and inter-site (Table S4.) variability was assessed as described in the respective legends. (PDF 128Â kb)

Published
2017
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39. Additional File 3

Author

Barabas, Sascha, Spindler, Theresa, Kiener, Richard, Tonar, Charlotte, Lugner, Tamara, Batzilla, Julia, Bendfeldt, Hanna, Rascle, Anne, Asbach, Benedikt, Wagner, Ralf, and Deml, Ludwig

Abstract

Gating strategy of flow cytometry analyses following surface marker (CD3, CD4, CD8, CD56) and intracellular IFN-γ staining. PBMC (4 replicates each) from six CMV-seropositive healthy donors were stimulated with T-activated® pp65 and IE-1 antigens for 6 h, as indicated in the legend to Fig. 9. (A) Cells were first gated on single events in a Forward Scatter-Area (FSC-A) x FSC-Height (FSC-H) dot plot (Gating #1). Based on FSC-A x Side Scatter-Area (SSC-A) properties, a gate was next set around lymphocytes (Gating #2). By plotting CD3 against CD56, CD56 single positive cells (Gating #3), CD3/CD56 double positive cells (Gating #4) and CD3 single positive cells (Gating #5) were separated. CD3 single positive cells (Gating #5) were further subdivided into CD3/CD4 double positive (Gating #6) and CD3/CD8 double positive cells (Gating #7). (B) Representative dot plots (1 out of 4 replicates) of stained PBMC of donor d290. In each plot, percentage value in gated area is the mean frequency (from 4 replicates) of IFN-γ-expressing CD3+CD4+ (Th), CD3+CD8+ (CTL), CD3−CD56+ (NK) and CD3+CD56+ (NKT-like) cells. (PDF 351 kb)

Published
2017
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40. Additional File 2

Author

Barabas, Sascha, Spindler, Theresa, Kiener, Richard, Tonar, Charlotte, Lugner, Tamara, Batzilla, Julia, Bendfeldt, Hanna, Rascle, Anne, Asbach, Benedikt, Wagner, Ralf, and Deml, Ludwig

Abstract

Positivity cut-off definition. Results obtained using the optimized IFN-γ ELISpot assay on PBMC isolated from whole blood of 45 healthy donors (Table 1 and Fig. 8) were used to generate precision profiles for each IE-1 and pp65 stimulation. Limits of quantitation (LoQ) for IE-1 and pp65 were 8.6 and 7.1 SFC/200,000 PBMC respectively. LoQ calculated on a cohort of 124 hemodialysis patients [49] were in a similar range (7.8 and 8.3 for IE-1 and pp65 respectively). Therefore, a positivity cut-off of 10 SFC/200,000 PBMC was chosen for the standardized assay. In addition, intra-assay standard deviation (SD) within both cohorts of healthy donors (n = 45) and hemodialysis patients (n = 124) for stimulated and unstimulated measurements was the basis for the calculation of a criterion that the ratio of geometric means of stimulated to unstimulated values is at least 2.5. Finally, considering a test result as positive when geometric mean for at least one of the IE-1 or pp65 stimulated approach is ≥ 10 SFC/200,000 PBMC, positive agreement (sensitivity) and negative agreement (specificity) of the optimized IFN-γ ELISpot test results with CMV serology within the collective of 45 healthy donors was 97% and 85% respectively (Fig. 8). (PDF 73 kb)

Published
2017
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41. Efficient Delivery of Human Cytomegalovirus T Cell Antigens by Attenuated Sendai Virus Vectors

Author

Kiener, Richard, primary, Fleischmann, Markus, additional, Wiegand, Marian Alexander, additional, Lemmermann, Niels A. W., additional, Schwegler, Christiane, additional, Kaufmann, Christine, additional, Renzaho, Angelique, additional, Thomas, Simone, additional, Felder, Eva, additional, Niller, Hans Helmut, additional, Asbach, Benedikt, additional, and Wagner, Ralf, additional

Published
2018
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45. An optimized IFN-γ ELISpot assay for the sensitive and standardized monitoring of CMV protein-reactive effector cells of cell-mediated immunity

Author

Barabas, Sascha, primary, Spindler, Theresa, additional, Kiener, Richard, additional, Tonar, Charlotte, additional, Lugner, Tamara, additional, Batzilla, Julia, additional, Bendfeldt, Hanna, additional, Rascle, Anne, additional, Asbach, Benedikt, additional, Wagner, Ralf, additional, and Deml, Ludwig, additional

Published
2017
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46. Replication-competent NYVAC-KC yields improved immunogenicity to HIV-1 antigens in rhesus macaques, compared to non-replicating NYVAC.

Author

Kibler, Karen V., Asbach, Benedikt, Perdiguero, Beatriz, García-Arriaza, Juan, Yates, Nicole L., Parks, Robert, Stanfield-Oakley, Sherry, Ferrari, Guido, Montefiori, David C., Tomaras, Georgia D., Roederer, Mario, Foulds, Kathryn E., Forthal, Donald N., Seaman, Michael S., Self, Steve, Gottardo, Raphael, Phogat, Sanjay, Tartaglia, James, Barnett, Susan, and Cristillo, Anthony D.

Subjects
*RHESUS monkeys, *IMMUNOGENETICS, *HIV infections, *AIDS vaccines, *POXVIRUSES
Abstract

As part of the continuing effort to develop an effective HIV vaccine, we generated a poxviral vaccine vector (previously described) designed to improve on the results of the RV144 Phase III clinical trial. The construct, NYVAC-KC, is a replication-competent, attenuated recombinant of the vaccinia virus strain, NYVAC. NYVAC is a vector that has been used in many previous clinical studies but is replication deficient. Here we report a side-by-side comparison of replication-restricted NYVAC and replication-competent NYVAC-KC in a non-human primate study, which utilized a prime-boost regimen similar to that of RV144. NYVAC-C and NYVAC-C-KC express the HIV-1 antigens gp140 and Gag/Gag-Pol-Nef-derived VLPs from clade C and were used as the prime, with recombinant virus plus envelope protein as the boost. In nearly every T and B cell immune assay against HIV-1, including neutralization and antibody binding, NYVAC-C-KC induced a greater immune response than did NYVAC-C, indicating that replication competence in a poxvirus may improve upon the modestly successful regimen used in the RV144 clinical trial. [ABSTRACT FROM AUTHOR]

Published
2019
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47. Proceedings of the Frontiers of Retrovirology Conference 2016

Author

Zurnic, Irena, primary, Hütter, Sylvia, additional, Lehmann, Ute, additional, Stanke, Nicole, additional, Reh, Juliane, additional, Kern, Tobias, additional, Lindel, Fabian, additional, Gerresheim, Gesche, additional, Hamann, Martin, additional, Müllers, Erik, additional, Lesbats, Paul, additional, Cherepanov, Peter, additional, Serrao, Erik, additional, Engelman, Alan, additional, Lindemann, Dirk, additional, Da Silva Santos, Claire, additional, Tartour, Kevin, additional, Cimarelli, Andrea, additional, Burdick, Rya, additional, Chen, Jianbo, additional, Sastri, Jaya, additional, Hu, Wei-Shau, additional, Pathak, Vinay, additional, Keppler, Oliver T., additional, Pradeau, Karine, additional, Eiler, Sylvia, additional, Levy, Nicolas, additional, Lennon, Sarah, additional, Cianferani, Sarah, additional, Emiliani, Stéphane, additional, Ruff, Marc, additional, Parissi, Vincent, additional, Rato, Sylvie, additional, Rausell, Antonio, additional, Munoz, Miguel, additional, Telenti, Amalio, additional, Ciuffi, Angela, additional, Zhyvoloup, Alexander, additional, Melamed, Anat, additional, Anderson, Ian, additional, Planas, Delphine, additional, Kriston-Vizi, Janos, additional, Ketteler, Robin, additional, Lee, Chen- Hsuin, additional, Merritt, Andy, additional, Ancuta, Petronela, additional, Bangham, Charles, additional, Fassati, Ariberto, additional, Rodari, Anthony, additional, Van Driessche, Benoit, additional, Galais, Mathilde, additional, Delacourt, Nadége, additional, Fauquenoy, Sylvain, additional, Vanhulle, Caroline, additional, Kula, Anna, additional, Burny, Arsène, additional, Rohr, Olivier, additional, Van Lint, Carine, additional, van Montfort, Thijs, additional, van der Sluis, Renee, additional, Speijer, Dave, additional, Berkhout, Ben, additional, Meng, Bo, additional, Rutkowski, Andrzej, additional, Berry, Neil, additional, Dölken, Lars, additional, Lever, Andrew, additional, Schuster, Thomas, additional, Asbach, Benedikt, additional, Wagner, Ralf, additional, Gross, Christine, additional, Wiesmann, Veit, additional, Kalmer, Martina, additional, Wittenberg, Thomas, additional, Gettemans, Jan, additional, Thoma-Kress, Andrea K., additional, Li, Minghua, additional, Freed, Eric O., additional, Liu, Shan-Lu, additional, Müller, Janis, additional, Münch, Jan, additional, Sewald, Xaver, additional, Uchil, Pradeep, additional, Ladinsky, Mark, additional, Beloor, Jagadish, additional, Pi, Ruoxi, additional, Herrmann, Christin, additional, Motamedi, Nasim, additional, Murooka, Thomas, additional, Brehm, Michael, additional, Greiner, Dale, additional, Mempel, Thorsten, additional, Bjorkman, Pamela, additional, Kumar, Priti, additional, Mothes, Walther, additional, Joas, Simone, additional, Parrish, Erica, additional, Gnanadurai, Clement Wesley, additional, Lump, Edina, additional, Stürzel, Christina M., additional, Parrish, Nicholas F., additional, Sauermann, Ulrike, additional, Töpfer, Katharina, additional, Schultheiss, Tina, additional, Bosinger, Steven, additional, Silvestri, Guido, additional, Apetrei, Cristian, additional, Huot, Nicholas, additional, Müller-Trutwin, Michaela, additional, Sauter, Daniel, additional, Hahn, Beatrice H., additional, Stahl-Hennig, Christiane, additional, Kirchhoff, Frank, additional, Schumann, Gerald, additional, Jung-Klawitter, Sabine, additional, Fuchs, Nina V., additional, Upton, Kyle R., additional, Muñoz-Lopez, Martin, additional, Shukla, Ruchi, additional, Wang, Jichang, additional, Garcia-Canadas, Marta, additional, Lopez-Ruiz, Cesar, additional, Gerhardt, Daniel J., additional, Sebe, Attila, additional, Grabundzija, Ivana, additional, Gerdes, Patricia, additional, Merkert, Sylvia, additional, Pulgarin, Andres, additional, Bock, Anja, additional, Held, Ulrike, additional, Witthuhn, Anett, additional, Haase, Alexandra, additional, Wolvetang, Ernst J., additional, Martin, Ulrich, additional, Ivics, Zoltán, additional, Izsvák, Zsuzsanna, additional, Garcia-Perez, J., additional, Faulkner, Geoffrey J., additional, Hurst, Tara, additional, Katzourakis, Aris, additional, Magiorkinis, Gkikas, additional, Schott, Kerstin, additional, Derua, Rita, additional, Seifried, Janna, additional, Reuter, Andreas, additional, Schmitz, Heike, additional, Tondera, Christiane, additional, Brandariz-Nuñez, Alberto, additional, Diaz-Griffero, Felipe, additional, Janssens, Veerle, additional, König, Renate, additional, Baldauf, Hanna-Mari, additional, Stegmann, Lena, additional, Schwarz, Sarah-Marie, additional, Trotard, Maud, additional, Martin, Margarethe, additional, Lenzi, Gina, additional, Burggraf, Manja, additional, Pan, Xiaoyu, additional, Fregoso, Oliver I., additional, Lim, Efrem S., additional, Abraham, Libin, additional, Erikson, Elina, additional, Nguyen, Laura, additional, Ambiel, Ina, additional, Rutsch, Frank, additional, Kim, Baek, additional, Emerman, Michael, additional, Fackler, Oliver T., additional, Wittmann, Sabine, additional, Behrendt, Rayk, additional, Volkmann, Bianca, additional, Eissmann, Kristin, additional, Gramberg, Thomas, additional, Bolduan, Sebastian, additional, Koppensteiner, Herwig, additional, Regensburg, Stefanie, additional, Brack-Werner, Ruth, additional, Draenert, Rika, additional, Schindler, Michael, additional, Ducroux, Aurélie, additional, Xu, Shuting, additional, Ponnurangam, Aparna, additional, Franz, Sergej, additional, Malassa, Angelina, additional, Ewald, Ellen, additional, Goffinet, Christine, additional, Fung, Sin-Yee, additional, Chan, Ching-Ping, additional, Yuen, Chun-Kit, additional, Kok, Kin-Hang, additional, Chan, Chin-Ping, additional, Jin, Dong-Yan, additional, Dittmer, Ulf, additional, Kmiec, Dorota, additional, Iyer, Shilpa, additional, Stürzel, Christina, additional, Hahn, Beatrice, additional, Ariumi, Yasuo, additional, Yasuda-Inoue, Mariko, additional, Kawano, Koudai, additional, Tateishi, Satoshi, additional, Turelli, Priscilla, additional, Compton, Alex, additional, Roy, Nicolas, additional, Porrot, Françoise, additional, Billet, Anne, additional, Casartelli, Nicoletta, additional, Yount, Jacob, additional, Liang, Chen, additional, Schwartz, Oliver, additional, Magnus, Carsten, additional, Reh, Lucia, additional, Moore, Penny, additional, Uhr, Therese, additional, Weber, Jacqueline, additional, Morris, Lynn, additional, Trkola, Alexandra, additional, Grindberg, Rashel V., additional, Schlaepfer, Erika, additional, Schreiber, Gideon, additional, Simon, Viviana, additional, Speck, Roberto F., additional, Debyser, Zeger, additional, Vranckx, Lenard, additional, Demeulemeester, Jonas, additional, Saleh, Suha, additional, Verdin, Eric, additional, Cereseto, Anna, additional, Christ, Frauke, additional, Gijsbers, Rik, additional, Wang, Gang, additional, Zhao, Na, additional, Das, Atze T., additional, Köstler, Josef, additional, Perdiguero, Beatriz, additional, Esteban, Mariano, additional, Jacobs, Bertram L., additional, Montefiori, David C., additional, LaBranche, Celia C., additional, Yates, Nicole L., additional, Tomaras, Georgia D., additional, Ferrari, Guido, additional, Foulds, Kathryn E., additional, Roederer, Mario, additional, Landucci, Gary, additional, Forthal, Donald N., additional, Seaman, Michael S., additional, Hawkins, Natalie, additional, Self, Steven G., additional, Phogat, Sanjay, additional, Tartaglia, James, additional, Barnett, Susan W., additional, Burke, Brian, additional, Cristillo, Anthony D., additional, Ding, Song, additional, Heeney, Jonathan L., additional, Pantaleo, Giuseppe, additional, Stab, Viktoria, additional, Ensser, Armin, additional, Tippler, Bettina, additional, Burton, Dennis, additional, Tenbusch, Matthias, additional, Überla, Klaus, additional, Alter, Galit, additional, Lofano, Giuseppe, additional, Dugast, Anne-Sophie, additional, Kulkarni, Viraj, additional, Suscovich, Todd, additional, Opazo, Tatiana, additional, Barraza, Felipe, additional, Herrera, Diego, additional, Garces, Andrea, additional, Schwenke, Tomas, additional, Tapia, Diego, additional, Cancino, Jorge, additional, Arriagada, Gloria, additional, Haußner, Christina, additional, Damm, Dominik, additional, Rohrhofer, Anette, additional, Schmidt, Barbara, additional, Eichler, Jutta, additional, Midgley, Rebecca, additional, Wheeldon, James, additional, Piguet, Vincent, additional, Khopkar, Priyanka, additional, Rohamare, Megha, additional, Kulkarni, Smita, additional, Godinho-Santos, Ana, additional, Hance, Allan, additional, Goncalves, Joao, additional, Mammano, Fabrizio, additional, Gasser, Romain, additional, Hamoudi, Meriem, additional, Pellicciotta, Martina, additional, Zhou, Zhicheng, additional, Visdeloup, Clara, additional, Colin, Philippe, additional, Braibant, Martine, additional, Lagane, Bernard, additional, Negroni, Matteo, additional, Wamara, Jula, additional, Bannert, Norbert, additional, Mesplede, Thibault, additional, Osman, Nathan, additional, Anstett, Kaitlin, additional, Liang, Jiaming Calvin, additional, Pham, Hanh Thi, additional, Wainberg, Mark, additional, Shao, Wei, additional, Shan, Jigui, additional, Kearney, Mary, additional, Wu, Xiaolin, additional, Maldarelli, Frank, additional, Mellors, John, additional, Luke, Brian, additional, Coffin, John, additional, Hughes, Stephen, additional, Fricke, Thomas, additional, Opp, Silvana, additional, Shepard, Caitlin, additional, Ivanov, Dmitri, additional, Valle-Casuso, Jose, additional, Kanja, Marine, additional, Cappy, Pierre, additional, Lener, Daniela, additional, Knyazhanskaya, Ekaterina, additional, Anisenko, Andrey, additional, Zatsepin, Timofey, additional, Gottikh, Marina, additional, Komkov, Alexander, additional, Minervina, Anastasia, additional, Nugmanov, Gaiaz, additional, Nazarov, Vadim, additional, Khodosevich, Konstantin, additional, Mamedov, Ilgar, additional, Lebedev, Yuri, additional, Colomer-Lluch, Marta, additional, Serra-Moreno, Ruth, additional, Sarracino, Ambra, additional, Gharu, Lavina, additional, Pasternak, Alexander, additional, Marcello, Alessandro, additional, McCartin, Ann Marie, additional, Kulkarni, Anurag, additional, Le Douce, Valentin, additional, Gautier, Virginie, additional, Baeyens, Ann, additional, Naessens, Evelien, additional, Van Nuffel, Anouk, additional, Weening, Karin, additional, Reilly, Anne- Marie, additional, Claeys, Eva, additional, Trypsteen, Wim, additional, Vandekerckhove, Linos, additional, Eyckerman, Sven, additional, Gevaert, Kris, additional, Verhasselt, Bruno, additional, Mok, Hoi Ping, additional, Norton, Nicholas, additional, Fun, Axel, additional, Hirst, Jack, additional, Wills, Mark, additional, Miklik, Dalibor, additional, Senigl, Filip, additional, Hejnar, Jiri, additional, Sakuragi, Jun-ichi, additional, Sakuragi, Sayuri, additional, Yokoyama, Masaru, additional, Shioda, Tatsuo, additional, Sato, Hironori, additional, Bodem, Jochen, additional, Moschall, Rebecca, additional, Denk, Sarah, additional, Erkelenz, Steffen, additional, Schenk, Christian, additional, Schaal, Heiner, additional, Donhauser, Norbert, additional, Socher, Ellen, additional, Millen, Sebastian, additional, Sticht, Heinrich, additional, Mann, Melanie, additional, Wei, Guochao, additional, Betts, Matthew J., additional, Liu, Yang, additional, Kehl, Timo, additional, Russell, Robert B., additional, Löchelt, Martin, additional, Hohn, Oliver, additional, Mostafa, Saeed, additional, Hanke, Kirsten, additional, Norley, Stephen, additional, Chen, Chia-Yen, additional, Shingai, Masashi, additional, Borrego, Pedro, additional, Taveira, Nuno, additional, Strebel, Klaus, additional, Hellmund, Chris, additional, Friedrich, Melanie, additional, Hahn, Friedrich, additional, Setz, Christian, additional, Rauch, Pia, additional, Fraedrich, Kirsten, additional, Matthaei, Alina, additional, Henklein, Petra, additional, Traxdorf, Maximilian, additional, Fossen, Torgils, additional, Schubert, Ulrich, additional, Khwaja, Aya, additional, Galilee, Meytal, additional, Alian, Akram, additional, Schwalbe, Birco, additional, Hauser, Heiko, additional, Schreiber, Michael, additional, Scherpenisse, Mirte, additional, Cho, Young-Keol, additional, Kim, Jungeun, additional, Jeong, Daeun, additional, Trejbalova, Katerina, additional, Benesova, Martina, additional, Kucerova, Dana, additional, Vernerova, Zdenka, additional, Amouroux, Rachel, additional, Hajkova, Petra, additional, Elleder, Daniel, additional, Hron, Tomas, additional, Farkasova, Helena, additional, Padhi, Abinash, additional, Paces, Jan, additional, Zhu, Henan, additional, Gifford, Robert, additional, Murcia, Pablo, additional, Carrozza, Maria Luisa, additional, Niewiadomska, Anna-Maria, additional, Mazzei, Maurizio, additional, Abi-Said, Mounir, additional, Hughes, Joseph, additional, Hué, Stéphane, additional, Obasa, Adetayo, additional, Jacobs, Graeme, additional, Engelbrecht, Susan, additional, Mack, Katharina, additional, Starz, Kathrin, additional, Geyer, Matthias, additional, Bibollet-Ruche, Frederic, additional, Leoz, Marie, additional, Plantier, Jean Christophe, additional, Argaw-Denboba, Ayele, additional, Balestrieri, Emanuela, additional, Serafino, Annalucia, additional, Bucci, Ilaria, additional, Cipriani, Chiara, additional, Spadafora, Corrado, additional, Sinibaldi-Vallebona, Paolo, additional, Matteucci, Claudia, additional, Jayashree, S. Nandi, additional, Neogi, Ujjwal, additional, Chhangani, Anil K., additional, Rathore, Shravan Sing, additional, Mathur, Bajrang R. J., additional, Abati, Adeyemi, additional, Koç, B. Taylan, additional, Oğuzoğlu, Tuba Çiğdem, additional, Shimauchi, Takatoshi, additional, Caucheteux, Stephan, additional, Turpin, Jocelyn, additional, Finsterbusch, Katja, additional, Tokura, Yoshiki, additional, Souriant, Shanti, additional, Balboa, Luciana, additional, Pingris, Karine, additional, Kviatcowsky, Denise, additional, Raynaud-Messina, Brigitte, additional, Cougoule, Céline, additional, Mercier, Ingrid, additional, Kuroda, Marcelo, additional, González-Montaner, Pablo, additional, Inwentarz, Sandra, additional, Moraña, Eduardo Jose, additional, del Carmen Sasiain, Maria, additional, Neyrolles, Olivier, additional, Maridonneau-Parini, Isabelle, additional, Lugo-Villarino, Geanncarlo, additional, Vérollet, Christel, additional, Herrmann, Alexandra, additional, Thomas, Dominique, additional, Bouzas, Nerea Ferreirós, additional, Lahaye, Xavier, additional, Bhargava, Anvita, additional, Satoh, Takeshi, additional, Gentili, Matteo, additional, Cerboni, Silvia, additional, Silvin, Aymeric, additional, Conrad, Cécile, additional, Ahmed-Belkacem, Hakim, additional, Rodriguez, Elisa C., additional, Guichou, Jean-François, additional, Bosquet, Nathalie, additional, Piel, Matthieu, additional, Le Grand, Roger, additional, King, Megan, additional, Pawlotsky, Jean-Michel, additional, Manel, Nicolas, additional, Hofmann, Henning, additional, Vanwalscappel, Benedicte, additional, Bloch, Nicolin, additional, Landau, Nathaniel, additional, Indik, Stanislav, additional, Hagen, Benedikt, additional, Valle-Casuso, José Carlos, additional, Allouch, Awatef, additional, David, Annie, additional, Barré-Sinoussi, Françoise, additional, Benkirane, Monsef, additional, Pancino, Gianfranco, additional, Saez-Cirion, Asier, additional, Lee, Wing-Yiu, additional, Sloan, Richard, additional, Schulte, Bianca, additional, Blomberg, Jonas, additional, Vargiu, Luana, additional, Rodriguez-Tomé, Patricia, additional, Tramontano, Enzo, additional, Sperber, Göran, additional, Kumari, Namita, additional, Ammosova, Tatiana, additional, Diaz, Sharmeen, additional, Oneal, Patricia, additional, Nekhai, Sergei, additional, Fahrny, Audrey, additional, Gers-Huber, Gustavo, additional, Audigé, Annette, additional, Jayaprakash, Anitha, additional, Sachidanandam, Ravi, additional, Hernandez, Matt, additional, Dillon-White, Marsha, additional, Maze, Emmanuel, additional, Ham, Claire, additional, Almond, Neil, additional, Towers, Greg, additional, Belshaw, Robert, additional, de Sousa-Pereira, Patrícia, additional, Abrantes, Joana, additional, Pizzato, Massimo, additional, Esteves, Pedro J., additional, Kahle, Tanja, additional, Schmitt, Sven, additional, Merkel, Laura, additional, Reuter, Nina, additional, Stamminger, Thomas, additional, Rosa, Ilaria Dalla, additional, Bishop, Kate, additional, Spinazzola, Antonella, additional, Groom, Harriet, additional, Vieyres, Gabrielle, additional, Müsken, Mathias, additional, Zillinger, Thomas, additional, Hornung, Veit, additional, Barchet, Winfried, additional, Häussler, Susanne, additional, Pietschmann, Thomas, additional, Javed, Aneela, additional, Leuchte, Nicole, additional, Salinas, Gabriela, additional, Opitz, Lennart, additional, Sopper, Sieghart, additional, Mummert, Christiane, additional, Hofmann, Christian, additional, Hückelhoven, Angela G., additional, Bergmann, Silke, additional, Müller-Schmucker, Sandra M., additional, Harrer, Ellen G., additional, Dörrie, Jan, additional, Schaft, Niels, additional, Harrer, Thomas, additional, Cardinaux, Laure, additional, Zahno, M.- L., additional, Vogt, H.- R., additional, Zanoni, R., additional, Bertoni, G., additional, Muenchhoff, Maximilian, additional, Goulder, Philip, additional, Keppler, Oliver, additional, Rebensburg, Stephanie, additional, Helfer, Markus, additional, Zhang, Yuwei, additional, Chen, Huicheng, additional, Bernier, Annie, additional, Gosselin, Annie, additional, Routy, Jean- Pierre, additional, Wöhrl, Birgitta, additional, Schneider, Anna, additional, Corona, Angela, additional, Spöring, Imke, additional, Jordan, Mareike, additional, Buchholz, Bernd, additional, Maccioni, Elias, additional, Di Santo, Roberto, additional, Schweimer, Kristian, additional, Schölz, Christian, additional, Weinert, Brian, additional, Wagner, Sebastian, additional, Beli, Petra, additional, Miyake, Yasuyuki, additional, Qi, Jun, additional, Jensen, Lars, additional, Streicher, Werner, additional, McCarthy, Anna, additional, Westwood, Nicholas, additional, Lain, Sonia, additional, Cox, Jürgen, additional, Matthias, Patrick, additional, Mann, Matthias, additional, Bradner, James, additional, Choudhary, Chunaram, additional, Stern, Marcel, additional, Valletta, Elena, additional, Frezza, Caterina, additional, Marino-Merlo, Francesca, additional, Grelli, Sandro, additional, Serafino, Anna Lucia, additional, Mastino, Antonio, additional, Macchi, Beatrice, additional, Kaulfuß, Meike, additional, Windmann, Sonja, additional, Bayer, Wibke, additional, Mikasi, Sello, additional, Heß, Rebecca, additional, Bonsmann, Michael Storcksdieck gen., additional, Kirschning, Carsten, additional, Lepenies, Bernd, additional, Kolenbrander, Anne, additional, Temchura, Vladimir, additional, Iijima, Kenta, additional, Kobayashi, Junya, additional, and Ishizaka, Yukihito, additional

Published
2016
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48. Targeting HIV-1 Env gp140 to LOX-1 Elicits Immune Responses in Rhesus Macaques

Author

Zurawski, Gerard, primary, Zurawski, Sandra, additional, Flamar, Anne-Laure, additional, Richert, Laura, additional, Wagner, Ralf, additional, Tomaras, Georgia D., additional, Montefiori, David C., additional, Roederer, Mario, additional, Ferrari, Guido, additional, Lacabaratz, Christine, additional, Bonnabau, Henri, additional, Klucar, Peter, additional, Wang, Zhiqing, additional, Foulds, Kathryn E., additional, Kao, Shing-Fen, additional, Yates, Nicole L., additional, LaBranche, Celia, additional, Jacobs, Bertram L., additional, Kibler, Karen, additional, Asbach, Benedikt, additional, Kliche, Alexander, additional, Salazar, Andres, additional, Reed, Steve, additional, Self, Steve, additional, Gottardo, Raphael, additional, Galmin, Lindsey, additional, Weiss, Deborah, additional, Cristillo, Anthony, additional, Thiebaut, Rodolphe, additional, Pantaleo, Giuseppe, additional, and Levy, Yves, additional

Published
2016
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49. Comprehensive analysis of interactions between the Src-associated protein in mitosis of 68 kDa and the human Src-homology 3 proteome

Author

Asbach, Benedikt, Ludwig, Christine, Saksela, Kalle, Wagner, Ralf, Haartman Institute (-2014), and Department of Virology

Subjects
STRUCTURAL BASIS, Proteomics, Proteome, Science, education, Blotting, Western, 610 Medizin, Enzyme-Linked Immunosorbent Assay, Biochemistry, Peptide Mapping, CELL-CYCLE PROGRESSION, Cell Line, src Homology Domains, DNA-binding proteins, Molecular Cell Biology, Macromolecular Structure Analysis, RNA-BINDING PROTEIN, Humans, ARGININE METHYLATION, SH3 DOMAINS, KH DOMAIN, Sorting Nexins, Biology, Adaptor Proteins, Signal Transducing, TYROSINE PHOSPHORYLATION, ddc:610, C-SRC, Mechanisms of Signal Transduction, Intracellular Signaling Peptides and Proteins, Membrane Proteins, RNA-Binding Proteins, Proteins, Protein interactions, Computational Biology, Regulatory proteins, INSULIN-RECEPTOR, KINASE HCK, Adaptor Proteins, Vesicular Transport, Cytoskeletal Proteins, Protein structure, Medicine, 3111 Biomedicine, Peptides, Protein Binding, Signal Transduction, Research Article
Abstract

The protein Sam68 is involved in many cellular processes such as cell-cycle regulation, RNA metabolism, or signal transduction. Sam68 comprises a central RNA-binding domain flanked by unstructured tails containing docking sites for signalling proteins including seven proline-rich sequences (denoted P0 to P6) as potential SH3-domain binding motifs. To comprehensively assess Sam68-SH3-interactions, we applied a phage-display screening of a library containing all approx. 300 human SH3 domains. Thereby we identified five new (from intersectin 2, the osteoclast stimulating factor OSF, nephrocystin, sorting nexin 9, and CIN85) and seven already known high-confidence Sam68-ligands (mainly from the Src-kinase family), as well as several lower-affinity binders. Interaction of the high-affinity Sam68-binders was confirmed in independent assays in vitro (phage-ELISA, GST-pull-down) and in vivo (FACS-based FRET-analysis with CFP- and YFP-tagged proteins). Fine-mapping analyses with peptides established P0, P3, P4, and P5 as exclusive docking-sites for SH3 domains, which showed varying preferences for these motifs. Mutational analyses identified individual residues within the proline-rich motifs being crucial for the interactions. Based on these data, we generated a Sam68-mutant incapable of interacting with SH3 domains any more, as subsequently demonstrated by FRET-analyses. In conclusion, we present a thorough characterization of Sam68’s interplay with the SH3 proteome. The observed interaction between Sam68 and OSF complements the known Sam68-Src and OSF-Src interactions. Thus, we propose, that Sam68 functions as a classical scaffold protein in this context, assembling components of an osteoclast-specific signalling pathway.

Published
2012

50. Head-to-Head Comparison of Poxvirus NYVAC and ALVAC Vectors Expressing Identical HIV-1 Clade C Immunogens in Prime-Boost Combination with Env Protein in Nonhuman Primates

Author

García-Arriaza, Juan, primary, Perdiguero, Beatriz, additional, Heeney, Jonathan, additional, Seaman, Michael, additional, Montefiori, David C., additional, Labranche, Celia, additional, Yates, Nicole L., additional, Shen, Xiaoying, additional, Tomaras, Georgia D., additional, Ferrari, Guido, additional, Foulds, Kathryn E., additional, McDermott, Adrian, additional, Kao, Shing-Fen, additional, Roederer, Mario, additional, Hawkins, Natalie, additional, Self, Steve, additional, Yao, Jiansheng, additional, Farrell, Patrick, additional, Phogat, Sanjay, additional, Tartaglia, Jim, additional, Barnett, Susan W., additional, Burke, Brian, additional, Cristillo, Anthony, additional, Weiss, Deborah, additional, Lee, Carter, additional, Kibler, Karen, additional, Jacobs, Bert, additional, Asbach, Benedikt, additional, Wagner, Ralf, additional, Ding, Song, additional, Pantaleo, Giuseppe, additional, and Esteban, Mariano, additional

Published
2015
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