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The molecular mechanism affecting translocation of newly synthesized herpesvirus nucleocapsids from the nucleus into the cytoplasm is still not fully understood. The viral nuclear egress complex (NEC) mediates budding at and scission from the inner nuclear membrane, but the NEC is not sufficient for efficient fusion of the primary virion envelope with the outer nuclear membrane. Since no other viral protein was found to be essential for this process, it was suggested that a cellular machinery is recruited by viral proteins. However, knowledge on fusion mechanisms involving the nuclear membranes is rare. Recently, vesicle-associated membrane protein-associated protein B (VAPB) was shown to play a role in nuclear egress of herpes simplex virus 1 (HSV-1). To test this for the related alphaherpesvirus pseudorabies virus (PrV), we mutated genes encoding VAPB and VAPA by CRISPR/Cas9-based genome editing in our standard rabbit kidney cells (RK13), either individually or in combination. Single as well as double knockout cells were tested for virus propagation and for defects in nuclear egress. However, no deficiency in virus replication nor any effect on nuclear egress was obvious suggesting that VAPB and VAPA do not play a significant role in this process during PrV infection in RK13 cells.
Anna Dorsch; Julia Hölper; Kati Franzke; Luca Zaeck; Thomas Mettenleiter; Barbara Klupp. Role of Vesicle-Associated Membrane Protein-Associated Proteins (VAP) A and VAPB in Nuclear Egress of the Alphaherpesvirus Pseudorabies Virus. Viruses 2021, 13, 1117 .
AMA StyleAnna Dorsch, Julia Hölper, Kati Franzke, Luca Zaeck, Thomas Mettenleiter, Barbara Klupp. Role of Vesicle-Associated Membrane Protein-Associated Proteins (VAP) A and VAPB in Nuclear Egress of the Alphaherpesvirus Pseudorabies Virus. Viruses. 2021; 13 (6):1117.
Chicago/Turabian StyleAnna Dorsch; Julia Hölper; Kati Franzke; Luca Zaeck; Thomas Mettenleiter; Barbara Klupp. 2021. "Role of Vesicle-Associated Membrane Protein-Associated Proteins (VAP) A and VAPB in Nuclear Egress of the Alphaherpesvirus Pseudorabies Virus." Viruses 13, no. 6: 1117.
The visualization of viral pathogens in infected tissues is an invaluable tool to understand spatial virus distribution, localization, and cell tropism in vivo. Commonly, virus-infected tissues are analyzed using conventional immunohistochemistry in paraffin-embedded thin sections. Here, we demonstrate the utility of volumetric three-dimensional (3D) immunofluorescence imaging using tissue optical clearing and light sheet microscopy to investigate host–pathogen interactions of pandemic SARS-CoV-2 in ferrets at a mesoscopic scale. The superior spatial context of large, intact samples (>150 mm3) allowed detailed quantification of interrelated parameters like focus-to-focus distance or SARS-CoV-2-infected area, facilitating an in-depth description of SARS-CoV-2 infection foci. Accordingly, we could confirm a preferential infection of the ferret upper respiratory tract by SARS-CoV-2 and suggest clustering of infection foci in close proximity. Conclusively, we present a proof-of-concept study for investigating critically important respiratory pathogens in their spatial tissue morphology and demonstrate the first specific 3D visualization of SARS-CoV-2 infection.
Luca Zaeck; David Scheibner; Julia Sehl; Martin Müller; Donata Hoffmann; Martin Beer; Elsayed Abdelwhab; Thomas Mettenleiter; ANGELE Breithaupt; Stefan Finke. Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model. Viruses 2021, 13, 529 .
AMA StyleLuca Zaeck, David Scheibner, Julia Sehl, Martin Müller, Donata Hoffmann, Martin Beer, Elsayed Abdelwhab, Thomas Mettenleiter, ANGELE Breithaupt, Stefan Finke. Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model. Viruses. 2021; 13 (3):529.
Chicago/Turabian StyleLuca Zaeck; David Scheibner; Julia Sehl; Martin Müller; Donata Hoffmann; Martin Beer; Elsayed Abdelwhab; Thomas Mettenleiter; ANGELE Breithaupt; Stefan Finke. 2021. "Light Sheet Microscopy-Assisted 3D Analysis of SARS-CoV-2 Infection in the Respiratory Tract of the Ferret Model." Viruses 13, no. 3: 529.
The live genetically-engineered oral rabies virus (RABV) variant SPBN GASGAS induces long-lasting immunity in foxes and protection against challenge with an otherwise lethal dose of RABV field strains both after experimental oral and parenteral routes of administration. Induction of RABV-specific binding antibodies and immunoglobulin isotypes (IgM, total IgG, IgG1, IgG2) were comparable in orally and parenterally vaccinated foxes. Differences were only observed in the induction of virus-neutralizing (VNA) titers, which were significantly higher in the parenterally vaccinated group. The dynamics of rabies-specific antibodies pre- and post-challenge (365 days post vaccination) suggest the predominance of type-1 immunity protection of SPBN GASGAS. Independent of the route of administration, in the absence of IgG1 the immune response to SPBN GAGAS was mainly IgG2 driven. Interestingly, vaccination with SPBN GASGAS does not cause significant differences in inducible IFN-γ production in vaccinated animals, indicating a relatively weak cellular immune response during challenge. Notably, the parenteral application of SPBN GASGAS did not induce any adverse side effects in foxes, thus supporting safety studies of this oral rabies vaccine in various species.
Verena Te Kamp; Virginia Friedrichs; Conrad Freuling; Ad Vos; Madlin Potratz; Antonia Klein; Luca Zaeck; Elisa Eggerbauer; Peter Schuster; Christian Kaiser; Steffen Ortmann; Antje Kretzschmar; Katharina Bobe; Michael Knittler; Anca Dorhoi; Stefan Finke; Thomas Müller. Comparable Long-Term Rabies Immunity in Foxes after IntraMuscular and Oral Application Using a Third-Generation Oral Rabies Virus Vaccine. Vaccines 2021, 9, 49 .
AMA StyleVerena Te Kamp, Virginia Friedrichs, Conrad Freuling, Ad Vos, Madlin Potratz, Antonia Klein, Luca Zaeck, Elisa Eggerbauer, Peter Schuster, Christian Kaiser, Steffen Ortmann, Antje Kretzschmar, Katharina Bobe, Michael Knittler, Anca Dorhoi, Stefan Finke, Thomas Müller. Comparable Long-Term Rabies Immunity in Foxes after IntraMuscular and Oral Application Using a Third-Generation Oral Rabies Virus Vaccine. Vaccines. 2021; 9 (1):49.
Chicago/Turabian StyleVerena Te Kamp; Virginia Friedrichs; Conrad Freuling; Ad Vos; Madlin Potratz; Antonia Klein; Luca Zaeck; Elisa Eggerbauer; Peter Schuster; Christian Kaiser; Steffen Ortmann; Antje Kretzschmar; Katharina Bobe; Michael Knittler; Anca Dorhoi; Stefan Finke; Thomas Müller. 2021. "Comparable Long-Term Rabies Immunity in Foxes after IntraMuscular and Oral Application Using a Third-Generation Oral Rabies Virus Vaccine." Vaccines 9, no. 1: 49.
There is a growing diversity of bat-associated lyssaviruses in the Old World. In August 2017, a dead Brandt’s bat (Myotis brandtii) tested positive for rabies and based on partial sequence analysis, the novel Kotalahti bat lyssavirus (KBLV) was identified. Because the bat was in an autolyzed state, isolation of KBLV was neither successful after three consecutive cell passages on cells nor in mice. Next generation sequencing (NGS) was applied using Ion Torrent ™ S5 technology coupled with target enrichment via hybridization-based capture (myBaits®) was used to sequence 99% of the genome, comprising of 11,878 nucleotides (nt). KBLV is most closely related to EBLV-2 (78.7% identity), followed by KHUV (79.0%) and BBLV (77.6%), supporting the assignment as phylogroup I lyssavirus. Interestingly, all of these lyssaviruses were also isolated from bat species of the genus Myotis, thus supporting that M. brandtii is likely the reservoir host. All information on antigenic and genetic divergence fulfil the species demarcation criteria by ICTV, so that we recommend KBLV as a novel species within the Lyssavirus genus. Next to sequence analyses, assignment to phylogroup I was functionally corroborated by cross-neutralization of G-deleted RABV, pseudotyped with KBLV-G by sera from RABV vaccinated humans. This suggests that conventional RABV vaccines also confer protection against the novel KBLV.
Sten Calvelage; Niina Tammiranta; Tiina Nokireki; Tuija Gadd; Elisa Eggerbauer; Luca M. Zaeck; Madlin Potratz; Claudia Wylezich; Dirk Höper; Thomas Müller; Stefan Finke; Conrad M. Freuling. Genetic and Antigenetic Characterization of the Novel Kotalahti Bat Lyssavirus (KBLV). Viruses 2021, 13, 69 .
AMA StyleSten Calvelage, Niina Tammiranta, Tiina Nokireki, Tuija Gadd, Elisa Eggerbauer, Luca M. Zaeck, Madlin Potratz, Claudia Wylezich, Dirk Höper, Thomas Müller, Stefan Finke, Conrad M. Freuling. Genetic and Antigenetic Characterization of the Novel Kotalahti Bat Lyssavirus (KBLV). Viruses. 2021; 13 (1):69.
Chicago/Turabian StyleSten Calvelage; Niina Tammiranta; Tiina Nokireki; Tuija Gadd; Elisa Eggerbauer; Luca M. Zaeck; Madlin Potratz; Claudia Wylezich; Dirk Höper; Thomas Müller; Stefan Finke; Conrad M. Freuling. 2021. "Genetic and Antigenetic Characterization of the Novel Kotalahti Bat Lyssavirus (KBLV)." Viruses 13, no. 1: 69.
Natural killer (NK) cells are critically involved in the early immune response against various intracellular pathogens, including Coxiella burnetii and Chlamydia psittaci . Chlamydia -infected NK cells functionally mature, induce cellular immunity, and protect themselves by killing the bacteria in secreted granules. Here, we report that infected NK cells do not allow intracellular multiday growth of Coxiella , as is usually observed in other host cell types.
Svea Matthiesen; Luca Zaeck; Kati Franzke; Rico Jahnke; Charlie Fricke; Michael Mauermeir; Stefan Finke; Anja Lührmann; Michael R. Knittler. Coxiella burnetii-Infected NK Cells Release Infectious Bacteria by Degranulation. Infection and Immunity 2020, 88, 1 .
AMA StyleSvea Matthiesen, Luca Zaeck, Kati Franzke, Rico Jahnke, Charlie Fricke, Michael Mauermeir, Stefan Finke, Anja Lührmann, Michael R. Knittler. Coxiella burnetii-Infected NK Cells Release Infectious Bacteria by Degranulation. Infection and Immunity. 2020; 88 (11):1.
Chicago/Turabian StyleSvea Matthiesen; Luca Zaeck; Kati Franzke; Rico Jahnke; Charlie Fricke; Michael Mauermeir; Stefan Finke; Anja Lührmann; Michael R. Knittler. 2020. "Coxiella burnetii-Infected NK Cells Release Infectious Bacteria by Degranulation." Infection and Immunity 88, no. 11: 1.
The visualization of viral pathogens in infected tissues is an invaluable tool to understand spatial virus distribution, localization, and cell tropism in vivo. Commonly, virus-infected tissues are analyzed using conventional immunohistochemistry in paraffin-embedded thin sections. Here, we demonstrate the utility of volumetric three-dimensional (3D) immunofluorescence imaging using tissue optical clearing and light sheet microscopy to investigate host-pathogen interactions of pandemic SARS-CoV-2 in ferrets at a mesoscopic scale. The superior spatial context of large, intact samples (> 150 mm3) allowed detailed quantification of interrelated parameters like focus-to-focus distance or SARS-CoV-2-infected area, facilitating an in-depth description of SARS-CoV-2 infection foci. Accordingly, we could confirm a preferential infection of the ferret upper respiratory tract by SARS-CoV-2 and emphasize a distinct focal infection pattern in nasal turbinates. Conclusively, we present a proof-of-concept study for investigating critically important respiratory pathogens in their spatial tissue morphology and demonstrate the first specific 3D visualization of SARS-CoV-2 infection.
Luca M. Zaeck; David Scheibner; Julia Sehl; Martin Müller; Donata Hoffmann; Martin Beer; Elsayed M. Abdelwhab; Thomas C. Mettenleiter; ANGELE Breithaupt; Stefan Finke. 3D reconstruction of SARS-CoV-2 infection in ferrets emphasizes focal infection pattern in the upper respiratory tract. 2020, 1 .
AMA StyleLuca M. Zaeck, David Scheibner, Julia Sehl, Martin Müller, Donata Hoffmann, Martin Beer, Elsayed M. Abdelwhab, Thomas C. Mettenleiter, ANGELE Breithaupt, Stefan Finke. 3D reconstruction of SARS-CoV-2 infection in ferrets emphasizes focal infection pattern in the upper respiratory tract. . 2020; ():1.
Chicago/Turabian StyleLuca M. Zaeck; David Scheibner; Julia Sehl; Martin Müller; Donata Hoffmann; Martin Beer; Elsayed M. Abdelwhab; Thomas C. Mettenleiter; ANGELE Breithaupt; Stefan Finke. 2020. "3D reconstruction of SARS-CoV-2 infection in ferrets emphasizes focal infection pattern in the upper respiratory tract." , no. : 1.
Although conventional immunohistochemistry for neurotropic rabies virus (RABV) usually shows high preference for neurons, non-neuronal cells are also potential targets, and abortive astrocyte infection is considered a main trigger of innate immunity in the CNS. While in vitro studies indicated differences between field and less virulent lab-adapted RABVs, a systematic, quantitative comparison of astrocyte tropism in vivo is lacking. Here, solvent-based tissue clearing was used to measure RABV cell tropism in infected brains. Immunofluorescence analysis of 1 mm-thick tissue slices enabled 3D-segmentation and quantification of astrocyte and neuron infection frequencies. Comparison of three highly virulent field virus clones from fox, dog, and raccoon with three lab-adapted strains revealed remarkable differences in the ability to infect astrocytes in vivo. While all viruses and infection routes led to neuron infection frequencies between 7–19%, striking differences appeared for astrocytes. Whereas astrocyte infection by field viruses was detected independent of the inoculation route (8–27%), only one lab-adapted strain infected astrocytes route-dependently [0% after intramuscular (i.m.) and 13% after intracerebral (i.c.) inoculation]. Two lab-adapted vaccine viruses lacked astrocyte infection altogether (0%, i.c. and i.m.). This suggests a model in which the ability to establish productive astrocyte infection in vivo functionally distinguishes field and attenuated lab RABV strains.
Madlin Potratz; Luca Zaeck; Michael Christen; Verena Te Kamp; Antonia Klein; Tobias Nolden; Conrad M. Freuling; Thomas Müller; Stefan Finke. Astrocyte Infection during Rabies Encephalitis Depends on the Virus Strain and Infection Route as Demonstrated by Novel Quantitative 3D Analysis of Cell Tropism. Cells 2020, 9, 412 .
AMA StyleMadlin Potratz, Luca Zaeck, Michael Christen, Verena Te Kamp, Antonia Klein, Tobias Nolden, Conrad M. Freuling, Thomas Müller, Stefan Finke. Astrocyte Infection during Rabies Encephalitis Depends on the Virus Strain and Infection Route as Demonstrated by Novel Quantitative 3D Analysis of Cell Tropism. Cells. 2020; 9 (2):412.
Chicago/Turabian StyleMadlin Potratz; Luca Zaeck; Michael Christen; Verena Te Kamp; Antonia Klein; Tobias Nolden; Conrad M. Freuling; Thomas Müller; Stefan Finke. 2020. "Astrocyte Infection during Rabies Encephalitis Depends on the Virus Strain and Infection Route as Demonstrated by Novel Quantitative 3D Analysis of Cell Tropism." Cells 9, no. 2: 412.
Although conventional immunohistochemistry for neurotropic Rabies virus (RABV) usually shows a high preference for neurons, non-neuronal cells are also potential target cells and abortive infection of astrocytes is considered a main trigger of innate immunity in the CNS. While in vitro studies indicated differences between field and less virulent lab-adapted RABVs, a systematic and quantitative comparison of astrocyte tropism in vivo is lacking. Here, a recently developed solvent-based tissue clearing technique was used to measure the RABV cell tropism in infected brains. Immunofluorescence analysis of 1 mm-thick tissue slices enabled 3D segmentation and quantification of infection frequencies of astrocytes and neurons. Comparison of highly virulent street virus clones from fox, dog, and raccoon with three lab strains of intermediate and low virulence revealed remarkable differences in the ability to infect astrocytes in vivo. While all viruses and infection routes led to comparable neuron infection frequencies, striking differences were detected for the infection of astrocytes. Consistent and inoculation route-independent astrocyte infection by field viruses, together with route-dependent or undetectable astrocyte infection by lab-adapted or vaccine viruses strongly suggests a model in which the ability to establish productive astrocyte infection in vivo functionally distinguishes field and attenuated lab RABV strains.
Madlin Potratz; Luca Zaeck; Michael Christen; Verena TeKamp; Antonia Klein; Tobias Nolden; Conrad M. Freuling; Thomas Müller; Stefan Finke. Astrocyte Infection during Rabies Encephalitis Depends on the Virus Strain and Infection Route as Demonstrated by Novel Quantitative 3D Analysis of Cell Tropism. 2020, 1 .
AMA StyleMadlin Potratz, Luca Zaeck, Michael Christen, Verena TeKamp, Antonia Klein, Tobias Nolden, Conrad M. Freuling, Thomas Müller, Stefan Finke. Astrocyte Infection during Rabies Encephalitis Depends on the Virus Strain and Infection Route as Demonstrated by Novel Quantitative 3D Analysis of Cell Tropism. . 2020; ():1.
Chicago/Turabian StyleMadlin Potratz; Luca Zaeck; Michael Christen; Verena TeKamp; Antonia Klein; Tobias Nolden; Conrad M. Freuling; Thomas Müller; Stefan Finke. 2020. "Astrocyte Infection during Rabies Encephalitis Depends on the Virus Strain and Infection Route as Demonstrated by Novel Quantitative 3D Analysis of Cell Tropism." , no. : 1.
The visualization of infection processes in relevant tissues and organs using microscopy methods reveals a unique link between the distribution, tropism, and abundance of pathogens and the physiological structure of the respective organ. To dissect virus replication and the host reaction in vivo at both a global and a single-cell level, conventional 2D imaging approaches can only provide limited insight. However, pathological studies of infected organ material are still mostly restricted to the immunostaining of thin sections from paraffin-embedded or frozen samples. While the 3D analysis of large tissue volumes is possible via laborious serial sectioning, a variety of problems and artifacts remain. Modern immunostaining-compatible tissue clearing techniques allow for the seamless 3D visualization of infection sites in optically cleared thick tissues sections or even entire organs. Benefiting from pure optical slicing, this approach enables the acquisition of multicolor high-volume 3D image stacks for coherent qualitative and quantitative analyses of the infection and its surrounding cellular environment. Here, we demonstrate the utility and power of this methodology by visualizing virus infections in different target tissues. For instance, we reconstructed the cellular context of rabies virus infection sites in mouse brain tissue, allowing a thorough investigation and quantitative analysis of rabies virus cell tropism. The systematic comparison of different rabies viruses with varying pathogenicity revealed a remarkable difference for highly virulent street rabies viruses and attenuated lab strains. While the virus protein expression was readily detectable at a comparable level in both neurons and non-neuronal glial cells from brains of mice infected with street rabies viruses, it was virtually absent in glial cells of lab strain-infected mice. These data provide novel and detailed insights into the pathogenesis of virus infections and substantially contribute to an improved understanding of virus–host interactions in vivo.
Luca Zaeck; Madlin Potratz; Antonia Klein; Conrad M. Freuling; Thomas Müller; Stefan Finke. High-Resolution 3D Imaging of Virus Infections in Solvent-Cleared Organs: Novel Insights into Virus Replication and Tropism In Vivo. Proceedings 2020, 50, 75 .
AMA StyleLuca Zaeck, Madlin Potratz, Antonia Klein, Conrad M. Freuling, Thomas Müller, Stefan Finke. High-Resolution 3D Imaging of Virus Infections in Solvent-Cleared Organs: Novel Insights into Virus Replication and Tropism In Vivo. Proceedings. 2020; 50 (1):75.
Chicago/Turabian StyleLuca Zaeck; Madlin Potratz; Antonia Klein; Conrad M. Freuling; Thomas Müller; Stefan Finke. 2020. "High-Resolution 3D Imaging of Virus Infections in Solvent-Cleared Organs: Novel Insights into Virus Replication and Tropism In Vivo." Proceedings 50, no. 1: 75.
Although most non-segmented negative-strand RNA viruses (NNSVs) replicate in the cytoplasm, NNSV proteins often exert host manipulatory functions in the nucleus. Matrix (M) proteins of henipaviruses and other paramyxoviruses shuttle through the nucleus, where host factors may bind for M modification or host-cell manipulation. Acidic leucine-rich nuclear phosphoprotein 32 family member B (ANP32B) is an interactor of Hendra and Nipah virus M. Both accumulate in the nucleus in an ANP32B-dependent manner. Here we demonstrate that the nuclear localization signal (NLS) of ANP32B is dispensable for HeV M binding. Specific purification of M-ANP32B but not of M-ANP32A complexes revealed that neither the negatively charged acidic nor the leucine-rich regions of ANP32 proteins per se mediate interactions with henipavirus M proteins. Whereas pneumovirus M did not interact with ANP32B, Newcastle disease virus (NDV, genus Avulavirus), Sendai virus (SeV, genus Respirovirus), Measles virus (MeV, genus Morbillivirus) and Canine distemper virus (CDV, genus Morbillivirus) M were able to form complexes with ANP32B. However, in contrast to NDV M and SeV M, which accumulated in the nucleus ANP32B dependently, both morbillivirus Ms did not accumulate in the nucleus, neither at ANP32B overexpression nor after nuclear protein export inhibition. These results indicate that intracellular compartmentalization of cytoplasmic morbillivirus M and nuclear ANP32B prevented an intracellular interaction. Overall, we provide evidence for a general ability of paramyxovirus M proteins to interact with ANP32B. This suggests a conserved, yet to be clarified mechanism might play a role in host manipulation and immune regulation in infected hosts.
Maria Günther; Anja Bauer; Martin Müller; Luca Zaeck; Stefan Finke. Interaction of host cellular factor ANP32B with matrix proteins of different paramyxoviruses. Journal of General Virology 2019, 101, jgv001362 -58.
AMA StyleMaria Günther, Anja Bauer, Martin Müller, Luca Zaeck, Stefan Finke. Interaction of host cellular factor ANP32B with matrix proteins of different paramyxoviruses. Journal of General Virology. 2019; 101 (1):jgv001362-58.
Chicago/Turabian StyleMaria Günther; Anja Bauer; Martin Müller; Luca Zaeck; Stefan Finke. 2019. "Interaction of host cellular factor ANP32B with matrix proteins of different paramyxoviruses." Journal of General Virology 101, no. 1: jgv001362-58.
: To evaluate the long-term immunogenicity of the live-attenuated, oral rabies vaccine SPBN GASGAS in a full good clinical practice (GCP) compliant study, forty-six (46) healthy, seronegative red foxes (Vulpes vulpes) were allocated to two treatment groups: group 1 (n = 31) received a vaccine bait containing 1.7 ml of the vaccine of minimum potency (106.6 FFU/mL) and group 2 (n = 15) received a placebo-bait. In total, 29 animals of group 1 and 14 animals of group 2 were challenged at 12 months post-vaccination with a fox rabies virus isolate (103.0 MICLD50/mL). While 90% of the animals offered a vaccine bait resisted the challenge, only one animal (7%) of the controls survived. All animals that had seroconverted following vaccination survived the challenge infection at 12 months post-vaccination. Rabies specific antibodies could be detected as early as 14 days post-vaccination. Based on the kinetics of the antibody response to SPBN GASGAS as measured in ELISA and RFFIT, the animals maintained stable antibody titres during the 12-month pre-challenge observation period at a high level. The results indicate that successful vaccination using the oral route with this new rabies virus vaccine strain confers long-term duration of immunity beyond one year, meeting the same requirements as for licensure as laid down by the European Pharmacopoeia.
Conrad M. Freuling; Verena Te Kamp; Antonia Klein; Maria Günther; Luca Zaeck; Madlin Potratz; Elisa Eggerbauer; Katharina Bobe; Christian Kaiser; Antje Kretzschmar; Steffen Ortmann; Peter Schuster; Adriaan Vos; Stefan Finke; Thomas Müller. Long-Term Immunogenicity and Efficacy of the Oral Rabies Virus Vaccine Strain SPBN GASGAS in Foxes. Viruses 2019, 11, 790 .
AMA StyleConrad M. Freuling, Verena Te Kamp, Antonia Klein, Maria Günther, Luca Zaeck, Madlin Potratz, Elisa Eggerbauer, Katharina Bobe, Christian Kaiser, Antje Kretzschmar, Steffen Ortmann, Peter Schuster, Adriaan Vos, Stefan Finke, Thomas Müller. Long-Term Immunogenicity and Efficacy of the Oral Rabies Virus Vaccine Strain SPBN GASGAS in Foxes. Viruses. 2019; 11 (9):790.
Chicago/Turabian StyleConrad M. Freuling; Verena Te Kamp; Antonia Klein; Maria Günther; Luca Zaeck; Madlin Potratz; Elisa Eggerbauer; Katharina Bobe; Christian Kaiser; Antje Kretzschmar; Steffen Ortmann; Peter Schuster; Adriaan Vos; Stefan Finke; Thomas Müller. 2019. "Long-Term Immunogenicity and Efficacy of the Oral Rabies Virus Vaccine Strain SPBN GASGAS in Foxes." Viruses 11, no. 9: 790.
The visualization of infection processes in tissues and organs by immunolabeling is a key method in modern infection biology. The ability to observe and study the distribution, tropism, and abundance of pathogens inside of organ tissues provides pivotal data on disease development and progression. Using conventional microscopy methods, immunolabeling is mostly restricted to thin sections obtained from paraffin-embedded or frozen samples. However, the limited 2D image plane of these thin sections may lead to the loss of crucial information on the complex structure of an infected organ and the cellular context of the infection. Modern multicolor, immunostaining-compatible tissue clearing techniques now provide a relatively fast and inexpensive way to study high-volume 3D image stacks of virus-infected organ tissue. By exposing the tissue to organic solvents, it becomes optically transparent. This matches the sample’s refractive indices and eventually leads to a significant reduction of light scattering. Thus, in combination with long free working distance objectives, large tissue sections up to 1 mm in size can be imaged by conventional confocal laser scanning microscopy (CLSM) at high resolution. Here, we describe a protocol to apply deep-tissue imaging after tissue clearing to visualize rabies virus distribution in infected brains in order to study topics like virus pathogenesis, spread, tropism, and neuroinvasion.
Luca Zaeck; Madlin Potratz; Conrad M. Freuling; Thomas Müller; Stefan Finke. High-Resolution 3D Imaging of Rabies Virus Infection in Solvent-Cleared Brain Tissue. Journal of Visualized Experiments 2019, e59402 .
AMA StyleLuca Zaeck, Madlin Potratz, Conrad M. Freuling, Thomas Müller, Stefan Finke. High-Resolution 3D Imaging of Rabies Virus Infection in Solvent-Cleared Brain Tissue. Journal of Visualized Experiments. 2019; (146):e59402.
Chicago/Turabian StyleLuca Zaeck; Madlin Potratz; Conrad M. Freuling; Thomas Müller; Stefan Finke. 2019. "High-Resolution 3D Imaging of Rabies Virus Infection in Solvent-Cleared Brain Tissue." Journal of Visualized Experiments , no. 146: e59402.