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Arthropod venoms offer a promising resource for the discovery of novel bioactive peptides and proteins, but the limited size of most species translates into minuscule venom yields. Bioactivity studies based on traditional fractionation are therefore challenging, so alternative strategies are needed. Cell-free synthesis based on synthetic gene fragments is one of the most promising emerging technologies, theoretically allowing the rapid, laboratory-scale production of specific venom components, but this approach has yet to be applied in venom biodiscovery. Here, we tested the ability of three commercially available cell-free protein expression systems to produce venom components from small arthropods, using U2-sicaritoxin-Sdo1a from the six-eyed sand spider Hexophtalma dolichocephala as a case study. We found that only one of the systems was able to produce an active product in low amounts, as demonstrated by SDS-PAGE, mass spectrometry, and bioactivity screening on murine neuroblasts. We discuss our findings in relation to the promises and limitations of cell-free synthesis for venom biodiscovery programs in smaller invertebrates.
Tim Lüddecke; Anne Paas; Lea Talmann; Kim N. Kirchhoff; Björn M. von Reumont; André Billion; Thomas Timm; Günter Lochnit; Andreas Vilcinskas. A Spider Toxin Exemplifies the Promises and Pitfalls of Cell-Free Protein Production for Venom Biodiscovery. Toxins 2021, 13, 575 .
AMA StyleTim Lüddecke, Anne Paas, Lea Talmann, Kim N. Kirchhoff, Björn M. von Reumont, André Billion, Thomas Timm, Günter Lochnit, Andreas Vilcinskas. A Spider Toxin Exemplifies the Promises and Pitfalls of Cell-Free Protein Production for Venom Biodiscovery. Toxins. 2021; 13 (8):575.
Chicago/Turabian StyleTim Lüddecke; Anne Paas; Lea Talmann; Kim N. Kirchhoff; Björn M. von Reumont; André Billion; Thomas Timm; Günter Lochnit; Andreas Vilcinskas. 2021. "A Spider Toxin Exemplifies the Promises and Pitfalls of Cell-Free Protein Production for Venom Biodiscovery." Toxins 13, no. 8: 575.
SARS-CoV-2 is a respiratory pathogen and primarily infects the airway epithelium. As our knowledge about innate immune factors of the respiratory tract against SARS-CoV-2 is limited, we generated and screened a peptide/protein library derived from bronchoalveolar lavage for inhibitors of SARS-CoV-2 spike-driven entry. Analysis of antiviral fractions revealed the presence of α1-antitrypsin (α1AT), a highly abundant circulating serine protease inhibitor. Here, we report that α1AT inhibits SARS-CoV-2 entry at physiological concentrations and suppresses viral replication in cell lines and primary cells including human airway epithelial cultures. We further demonstrate that α1AT binds and inactivates the serine protease TMPRSS2, which enzymatically primes the SARS-CoV-2 spike protein for membrane fusion. Thus, the acute phase protein α1AT is an inhibitor of TMPRSS2 and SARS-CoV-2 entry, and may play an important role in the innate immune defense against the novel coronavirus. Our findings suggest that repurposing of α1AT-containing drugs has prospects for the therapy of COVID-19.
Lukas Wettstein; Tatjana Weil; Carina Conzelmann; Janis A. Müller; Rüdiger Groß; Maximilian Hirschenberger; Alina Seidel; Susanne Klute; Fabian Zech; Caterina Prelli Bozzo; Nico Preising; Giorgio Fois; Robin Lochbaum; Philip Maximilian Knaff; Volker Mailänder; Ludger Ständker; Dietmar Rudolf Thal; Christian Schumann; Steffen Stenger; Alexander Kleger; Günter Lochnit; Benjamin Mayer; Yasser B. Ruiz-Blanco; Markus Hoffmann; Konstantin M. J. Sparrer; Stefan Pöhlmann; Elsa Sanchez-Garcia; Frank Kirchhoff; Manfred Frick; Jan Münch. Alpha-1 antitrypsin inhibits TMPRSS2 protease activity and SARS-CoV-2 infection. Nature Communications 2021, 12, 1 -10.
AMA StyleLukas Wettstein, Tatjana Weil, Carina Conzelmann, Janis A. Müller, Rüdiger Groß, Maximilian Hirschenberger, Alina Seidel, Susanne Klute, Fabian Zech, Caterina Prelli Bozzo, Nico Preising, Giorgio Fois, Robin Lochbaum, Philip Maximilian Knaff, Volker Mailänder, Ludger Ständker, Dietmar Rudolf Thal, Christian Schumann, Steffen Stenger, Alexander Kleger, Günter Lochnit, Benjamin Mayer, Yasser B. Ruiz-Blanco, Markus Hoffmann, Konstantin M. J. Sparrer, Stefan Pöhlmann, Elsa Sanchez-Garcia, Frank Kirchhoff, Manfred Frick, Jan Münch. Alpha-1 antitrypsin inhibits TMPRSS2 protease activity and SARS-CoV-2 infection. Nature Communications. 2021; 12 (1):1-10.
Chicago/Turabian StyleLukas Wettstein; Tatjana Weil; Carina Conzelmann; Janis A. Müller; Rüdiger Groß; Maximilian Hirschenberger; Alina Seidel; Susanne Klute; Fabian Zech; Caterina Prelli Bozzo; Nico Preising; Giorgio Fois; Robin Lochbaum; Philip Maximilian Knaff; Volker Mailänder; Ludger Ständker; Dietmar Rudolf Thal; Christian Schumann; Steffen Stenger; Alexander Kleger; Günter Lochnit; Benjamin Mayer; Yasser B. Ruiz-Blanco; Markus Hoffmann; Konstantin M. J. Sparrer; Stefan Pöhlmann; Elsa Sanchez-Garcia; Frank Kirchhoff; Manfred Frick; Jan Münch. 2021. "Alpha-1 antitrypsin inhibits TMPRSS2 protease activity and SARS-CoV-2 infection." Nature Communications 12, no. 1: 1-10.
Nemerteans (ribbon worms) employ toxins to subdue their prey, but research thus far has focused on the small-molecule components of mucus secretions and few protein toxins have been characterized. We carried out a preliminary proteotranscriptomic analysis of putative toxins produced by the hoplonemertean Amphiporus lactifloreus (Hoplonemertea, Amphiporidae). No variants were found of known nemertean-specific toxin proteins (neurotoxins, cytotoxins, parbolysins or nemertides) but several toxin-like transcripts were discovered, expressed strongly in the proboscis, including putative metalloproteinases and sequences resembling sea anemone actitoxins, crown-of-thorn sea star plancitoxins, and multiple classes of inhibitor cystine knot/knottin family proteins. Some of these products were also directly identified in the mucus proteome, supporting their preliminary identification as secreted toxin components. Two new nemertean-typical toxin candidates could be described and were named U-nemertotoxin-1 and U-nemertotoxin-2. Our findings provide insight into the largely overlooked venom system of nemerteans and support a hypothesis in which the nemertean proboscis evolved in several steps from a flesh-melting organ in scavenging nemerteans to a flesh-melting and toxin-secreting venom apparatus in hunting hoplonemerteans.
Björn Marcus Von Reumont; Tim Lüddecke; Thomas Timm; Günter Lochnit; Andreas Vilcinskas; Jörn Von Döhren; Maria A. Nilsson. Proteo-Transcriptomic Analysis Identifies Potential Novel Toxins Secreted by the Predatory, Prey-Piercing Ribbon Worm Amphiporus lactifloreus. Marine Drugs 2020, 18, 407 .
AMA StyleBjörn Marcus Von Reumont, Tim Lüddecke, Thomas Timm, Günter Lochnit, Andreas Vilcinskas, Jörn Von Döhren, Maria A. Nilsson. Proteo-Transcriptomic Analysis Identifies Potential Novel Toxins Secreted by the Predatory, Prey-Piercing Ribbon Worm Amphiporus lactifloreus. Marine Drugs. 2020; 18 (8):407.
Chicago/Turabian StyleBjörn Marcus Von Reumont; Tim Lüddecke; Thomas Timm; Günter Lochnit; Andreas Vilcinskas; Jörn Von Döhren; Maria A. Nilsson. 2020. "Proteo-Transcriptomic Analysis Identifies Potential Novel Toxins Secreted by the Predatory, Prey-Piercing Ribbon Worm Amphiporus lactifloreus." Marine Drugs 18, no. 8: 407.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19). To identify factors of the respiratory tract that suppress SARS-CoV-2, we screened a peptide/protein library derived from bronchoalveolar lavage, and identified α1-antitrypsin (α1-AT) as specific inhibitor of SARS-CoV-2. α1-AT targets the viral spike protein and blocks SARS-CoV-2 infection of human airway epithelium at physiological concentrations. Our findings show that endogenous α1-AT restricts SARS-CoV-2 and repurposes α1-AT-based drugs for COVID-19 therapy.
Lukas Wettstein; Carina Conzelmann; Janis A. Müller; Tatjana Weil; Ruediger Gross; Maximilian Hirschenberger; Alina Seidel; Susanne Klute; Fabian Zech; Caterina Prelli Bozzo; Nico Preising; Giorgio Fois; Robin Lochbaum; Philip Knaff; Volker Mailaender; Ludger Staendker; Dietmar Rudolf Thal; Christian Schumann; Steffen Stenger; Alexander Kleger; Guenter Lochnit; Konstantin Sparrer; Frank Kirchhoff; Manfred Frick; Jan Muench. Alpha-1 antitrypsin inhibits SARS-CoV-2 infection. 2020, 1 .
AMA StyleLukas Wettstein, Carina Conzelmann, Janis A. Müller, Tatjana Weil, Ruediger Gross, Maximilian Hirschenberger, Alina Seidel, Susanne Klute, Fabian Zech, Caterina Prelli Bozzo, Nico Preising, Giorgio Fois, Robin Lochbaum, Philip Knaff, Volker Mailaender, Ludger Staendker, Dietmar Rudolf Thal, Christian Schumann, Steffen Stenger, Alexander Kleger, Guenter Lochnit, Konstantin Sparrer, Frank Kirchhoff, Manfred Frick, Jan Muench. Alpha-1 antitrypsin inhibits SARS-CoV-2 infection. . 2020; ():1.
Chicago/Turabian StyleLukas Wettstein; Carina Conzelmann; Janis A. Müller; Tatjana Weil; Ruediger Gross; Maximilian Hirschenberger; Alina Seidel; Susanne Klute; Fabian Zech; Caterina Prelli Bozzo; Nico Preising; Giorgio Fois; Robin Lochbaum; Philip Knaff; Volker Mailaender; Ludger Staendker; Dietmar Rudolf Thal; Christian Schumann; Steffen Stenger; Alexander Kleger; Guenter Lochnit; Konstantin Sparrer; Frank Kirchhoff; Manfred Frick; Jan Muench. 2020. "Alpha-1 antitrypsin inhibits SARS-CoV-2 infection." , no. : 1.
Spiders use venom to subdue their prey, but little is known about the diversity of venoms in different spider families. Given the limited data available for orb-weaver spiders (Araneidae), we selected the wasp spider Argiope bruennichi for detailed analysis. Our strategy combined a transcriptomics pipeline based on multiple assemblies with a dual proteomics workflow involving parallel mass spectrometry techniques and electrophoretic profiling. We found that the remarkably simple venom of A. bruennichi has an atypical composition compared to other spider venoms, prominently featuring members of the cysteine-rich secretory protein, antigen 5 and pathogenesis-related protein 1 (CAP) superfamily and other, mostly high-molecular-weight proteins. We also detected a subset of potentially novel toxins similar to neuropeptides. We discuss the potential function of these proteins in the context of the unique hunting behavior of wasp spiders, which rely mostly on silk to trap their prey. We propose that the simplicity of the venom evolved to solve an economic dilemma between two competing yet metabolically expensive weapon systems. This study emphasizes the importance of cutting-edge methods to encompass the lineages of smaller venomous species that have yet to be characterized in detail, allowing us to understand the biology of their venom systems and to mine this prolific resource for translational research.
Tim Lüddecke; Björn M. Von Reumont; Frank Förster; André Billion; Thomas Timm; Günter Lochnit; Andreas Vilcinskas; Sarah Lemke. An Economic Dilemma Between Molecular Weapon Systems May Explain an Arachno-atypical Venom in Wasp Spiders (Argiope bruennichi). Biomolecules 2020, 10, 978 .
AMA StyleTim Lüddecke, Björn M. Von Reumont, Frank Förster, André Billion, Thomas Timm, Günter Lochnit, Andreas Vilcinskas, Sarah Lemke. An Economic Dilemma Between Molecular Weapon Systems May Explain an Arachno-atypical Venom in Wasp Spiders (Argiope bruennichi). Biomolecules. 2020; 10 (7):978.
Chicago/Turabian StyleTim Lüddecke; Björn M. Von Reumont; Frank Förster; André Billion; Thomas Timm; Günter Lochnit; Andreas Vilcinskas; Sarah Lemke. 2020. "An Economic Dilemma Between Molecular Weapon Systems May Explain an Arachno-atypical Venom in Wasp Spiders (Argiope bruennichi)." Biomolecules 10, no. 7: 978.
Activated caspase-1 and caspase-11 induce inflammatory cell death in a process termed pyroptosis. Here we show that Prostaglandin E2 (PGE2) inhibits caspase-11-dependent pyroptosis in murine and human macrophages. PGE2 suppreses caspase-11 expression in murine and human macrophages and in the airways of mice with allergic inflammation. Remarkably, caspase-11-deficient mice are strongly resistant to developing experimental allergic airway inflammation, where PGE2 is known to be protective. Expression of caspase-11 is elevated in the lung of wild type mice with allergic airway inflammation. Blocking PGE2 production with indomethacin enhances, whereas the prostaglandin E1 analog misoprostol inhibits lung caspase-11 expression. Finally, alveolar macrophages from asthma patients exhibit increased expression of caspase-4, a human homologue of caspase-11. Our findings identify PGE2 as a negative regulator of caspase-11-driven pyroptosis and implicate caspase-4/11 as a critical contributor to allergic airway inflammation, with implications for pathophysiology of asthma.
Zbigniew Zasłona; Ewelina Flis; Mieszko M. Wilk; Richard G. Carroll; Eva M. Palsson-McDermott; Mark M. Hughes; Ciana Diskin; Kathy Banahan; Dylan G. Ryan; Alexander Hooftman; Alicja Misiak; Jay Kearney; Gunter Lochnit; Wilhelm Bertrams; Timm Greulich; Bernd Schmeck; Oliver J. McElvaney; Kingston H. G. Mills; Ed C. Lavelle; Małgorzata Wygrecka; Emma M. Creagh; Luke A. J. O’Neill. Caspase-11 promotes allergic airway inflammation. Nature Communications 2020, 11, 1 -11.
AMA StyleZbigniew Zasłona, Ewelina Flis, Mieszko M. Wilk, Richard G. Carroll, Eva M. Palsson-McDermott, Mark M. Hughes, Ciana Diskin, Kathy Banahan, Dylan G. Ryan, Alexander Hooftman, Alicja Misiak, Jay Kearney, Gunter Lochnit, Wilhelm Bertrams, Timm Greulich, Bernd Schmeck, Oliver J. McElvaney, Kingston H. G. Mills, Ed C. Lavelle, Małgorzata Wygrecka, Emma M. Creagh, Luke A. J. O’Neill. Caspase-11 promotes allergic airway inflammation. Nature Communications. 2020; 11 (1):1-11.
Chicago/Turabian StyleZbigniew Zasłona; Ewelina Flis; Mieszko M. Wilk; Richard G. Carroll; Eva M. Palsson-McDermott; Mark M. Hughes; Ciana Diskin; Kathy Banahan; Dylan G. Ryan; Alexander Hooftman; Alicja Misiak; Jay Kearney; Gunter Lochnit; Wilhelm Bertrams; Timm Greulich; Bernd Schmeck; Oliver J. McElvaney; Kingston H. G. Mills; Ed C. Lavelle; Małgorzata Wygrecka; Emma M. Creagh; Luke A. J. O’Neill. 2020. "Caspase-11 promotes allergic airway inflammation." Nature Communications 11, no. 1: 1-11.
Monomeric serum immunoglobulin A (IgA) can contribute to the development of various autoimmune diseases, but the regulation of serum IgA effector functions is not well defined. Here, we show that the two IgA subclasses (IgA1 and IgA2) differ in their effect on immune cells due to distinct binding and signaling properties. Whereas IgA2 acts pro-inflammatory on neutrophils and macrophages, IgA1 does not have pronounced effects. Moreover, IgA1 and IgA2 have different glycosylation profiles, with IgA1 possessing more sialic acid than IgA2. Removal of sialic acid increases the pro-inflammatory capacity of IgA1, making it comparable to IgA2. Of note, disease-specific autoantibodies in patients with rheumatoid arthritis display a shift toward the pro-inflammatory IgA2 subclass, which is associated with higher disease activity. Taken together, these data demonstrate that IgA effector functions depend on subclass and glycosylation, and that disturbances in subclass balance are associated with autoimmune disease.
Ulrike Steffen; Carolien A. Koeleman; Maria V. Sokolova; Holger Bang; Arnd Kleyer; Jürgen Rech; Harald Unterweger; Martin Schicht; Fabian Garreis; Jonas Hahn; Fabian T. Andes; Fabian Hartmann; Madelaine Hahn; Aparna Mahajan; Friedrich Paulsen; Markus Hoffmann; Günter Lochnit; Luis E. Muñoz; Manfred Wuhrer; David Falck; Martin Herrmann; Georg Schett. IgA subclasses have different effector functions associated with distinct glycosylation profiles. Nature Communications 2020, 11, 1 -12.
AMA StyleUlrike Steffen, Carolien A. Koeleman, Maria V. Sokolova, Holger Bang, Arnd Kleyer, Jürgen Rech, Harald Unterweger, Martin Schicht, Fabian Garreis, Jonas Hahn, Fabian T. Andes, Fabian Hartmann, Madelaine Hahn, Aparna Mahajan, Friedrich Paulsen, Markus Hoffmann, Günter Lochnit, Luis E. Muñoz, Manfred Wuhrer, David Falck, Martin Herrmann, Georg Schett. IgA subclasses have different effector functions associated with distinct glycosylation profiles. Nature Communications. 2020; 11 (1):1-12.
Chicago/Turabian StyleUlrike Steffen; Carolien A. Koeleman; Maria V. Sokolova; Holger Bang; Arnd Kleyer; Jürgen Rech; Harald Unterweger; Martin Schicht; Fabian Garreis; Jonas Hahn; Fabian T. Andes; Fabian Hartmann; Madelaine Hahn; Aparna Mahajan; Friedrich Paulsen; Markus Hoffmann; Günter Lochnit; Luis E. Muñoz; Manfred Wuhrer; David Falck; Martin Herrmann; Georg Schett. 2020. "IgA subclasses have different effector functions associated with distinct glycosylation profiles." Nature Communications 11, no. 1: 1-12.
Within mega-diverse Hymenoptera, non-aculeate parasitic wasps represent 75% of all hymenopteran species. Their ovipositor dual-functionally injects venom and employs eggs into (endoparasitoids) or onto (ectoparasitoids) diverse host species. Few endoparasitoid wasps such as Pimpla turionellae paralyze the host and suppress its immune responses, such as encapsulation and melanization, to guarantee their offspring’s survival. Here, the venom and its possible biology and function of P. turionellae are characterized in comparison to the few existing proteo-transcriptomic analyses on parasitoid wasp venoms. Multiple transcriptome assembly and custom-tailored search and annotation strategies were applied to identify parasitoid venom proteins. To avoid false-positive hits, only transcripts were finally discussed that survived strict filter settings, including the presence in the proteome and higher expression in the venom gland. P. turionella features a venom that is mostly composed of known, typical parasitoid enzymes, cysteine-rich peptides, and other proteins and peptides. Several venom proteins were identified and named, such as pimplin2, 3, and 4. However, the specification of many novel candidates remains difficult, and annotations ambiguous. Interestingly, we do not find pimplin, a paralytic factor in Pimpla hypochondriaca, but instead a new cysteine inhibitor knot (ICK) family (pimplin2), which is highly similar to known, neurotoxic asilid1 sequences from robber flies.
Rabia Özbek; Natalie Wielsch; Heiko Vogel; Günter Lochnit; Frank Förster; Andreas Vilcinskas; Björn Marcus Von Reumont. Proteo-Transcriptomic Characterization of the Venom from the Endoparasitoid Wasp Pimpla turionellae with Aspects on Its Biology and Evolution. Toxins 2019, 11, 721 .
AMA StyleRabia Özbek, Natalie Wielsch, Heiko Vogel, Günter Lochnit, Frank Förster, Andreas Vilcinskas, Björn Marcus Von Reumont. Proteo-Transcriptomic Characterization of the Venom from the Endoparasitoid Wasp Pimpla turionellae with Aspects on Its Biology and Evolution. Toxins. 2019; 11 (12):721.
Chicago/Turabian StyleRabia Özbek; Natalie Wielsch; Heiko Vogel; Günter Lochnit; Frank Förster; Andreas Vilcinskas; Björn Marcus Von Reumont. 2019. "Proteo-Transcriptomic Characterization of the Venom from the Endoparasitoid Wasp Pimpla turionellae with Aspects on Its Biology and Evolution." Toxins 11, no. 12: 721.
Patients with dementia are increasing steadily, cognitive impairment by dementia not only exclusively suffers by old people but also young to middle aged individuals. However, the mechanism of cognitive impairment occurs in young people is not understood. Further, current medication to impairment did not provide satisfactory results. Therefore, we investigated the potential role of Ocimum sanctum ethanolic extract to enhance cognitive ability in the rat in vivo model. Young to middle aged rats were divided into 3 groups (3, 6, 9 months old) were treated with (0, 50 and 100 mg/kg b.w.) O. sanctum for 45 days. We employed a behavioral assay to assess cognitive ability. Further, Nissl staining was performed to analyze hippocampus formation in dentate gyrus (DG), cornu ammonis 1 (CA1), cornu ammonis 3 (CA3). The expression and activity of ChAT in brain was analyzed by RT-PCR and ELISA. Our results showed that treatment of O. sanctum with a dosage of 100 mg/kg b.w. for 45 days induced the cognitive ability in nine months old rats. Further, we observed a significant increase in density of granular and pyramidal cells in DG, CA1, and CA3. These results were corroborated by an increase in the ChAT activity and gene expression in the rat model as well as HEK 293 cell culture model. Taken together, the administration of 100 mg/kg b.w. O.sanctum induced the expression of ChAT. The increased ChAT expression and activity may enhance the cognitive ability in 9 months old rats mimicking young and middle aged condition in humans.
Dwi Liliek Kusindarta; Hevi Wihadmadyatami; Arvendi R. Jadi; Srikanth Karnati; Guenter Lochnit; Puspa Hening; Aris Haryanto; Made B. Auriva; Medania Purwaningrum. Ethanolic extract Ocimum sanctum. Enhances cognitive ability from young adulthood to middle aged mediated by increasing choline acetyl transferase activity in rat model. Research in Veterinary Science 2018, 118, 431 -438.
AMA StyleDwi Liliek Kusindarta, Hevi Wihadmadyatami, Arvendi R. Jadi, Srikanth Karnati, Guenter Lochnit, Puspa Hening, Aris Haryanto, Made B. Auriva, Medania Purwaningrum. Ethanolic extract Ocimum sanctum. Enhances cognitive ability from young adulthood to middle aged mediated by increasing choline acetyl transferase activity in rat model. Research in Veterinary Science. 2018; 118 ():431-438.
Chicago/Turabian StyleDwi Liliek Kusindarta; Hevi Wihadmadyatami; Arvendi R. Jadi; Srikanth Karnati; Guenter Lochnit; Puspa Hening; Aris Haryanto; Made B. Auriva; Medania Purwaningrum. 2018. "Ethanolic extract Ocimum sanctum. Enhances cognitive ability from young adulthood to middle aged mediated by increasing choline acetyl transferase activity in rat model." Research in Veterinary Science 118, no. : 431-438.
Proteins and glycolipids have been found to be decorated with phosphorylcholine (PC) both in protozoa and nematodes that parasitize humans and animals. PC epitopes can provoke various effects on immune cells leading to an immunomodulation of the host’s immune system that allows long-term persistence of the parasites. So far, only a limited number of PC-modified proteins, mainly from nematodes, have been identified. Infections caused by Leishmania spp. (e.g., L. infantum in southern Europe) affect about 12 million people worldwide and are characterized by a wide spectrum of clinical forms in humans, ranging from cutaneous to fatal visceral leishmaniasis. To establish and maintain the infection, these protozoa are dependent on the secretion of effector molecules into the host for modulating their immune system. In this project, we analyzed the PC modification of L. infantum promastigotes by 2D-gel based proteomics. Western blot analysis with the PC-specific antibody TEPC-15 revealed one PC-substituted protein in this organism, identified as eEF1α. We could demonstrate that the binding of eEF1α to one of its downstream effectors is dependent on its PC-modification. In this study we provide evidence that in this parasite the modification of eEF1α with PC may be essential for its function as an important virulence factor.
Thomas Timm; Giada Annoscia; Jochen Klein; Günter Lochnit. The Eukaryotic Elongation Factor 1 Alpha (eEF1α) from the Parasite Leishmania infantum Is Modified with the Immunomodulatory Substituent Phosphorylcholine (PC). Molecules 2017, 22, 2094 .
AMA StyleThomas Timm, Giada Annoscia, Jochen Klein, Günter Lochnit. The Eukaryotic Elongation Factor 1 Alpha (eEF1α) from the Parasite Leishmania infantum Is Modified with the Immunomodulatory Substituent Phosphorylcholine (PC). Molecules. 2017; 22 (12):2094.
Chicago/Turabian StyleThomas Timm; Giada Annoscia; Jochen Klein; Günter Lochnit. 2017. "The Eukaryotic Elongation Factor 1 Alpha (eEF1α) from the Parasite Leishmania infantum Is Modified with the Immunomodulatory Substituent Phosphorylcholine (PC)." Molecules 22, no. 12: 2094.
The coding sequence of a peroxidase from the secretome of Pleurotus sapidus was cloned from a cDNA library. Bioinformatic analyses revealed an open reading frame of 1551 bp corresponding to a primary translation product of 516 amino acids. The DyP-type peroxidase was heterologously produced in Trichoderma reesei with an activity of 55,000 U L−1. The enzyme was purified from the culture supernatant, biochemically characterized and the kinetic parameters were determined. The enzyme has an N-terminal signal peptide composed of 62 amino acids. Analysis by Blue Native PAGE and activity staining with ABTS, as well as gel filtration chromatography showed the native dimeric state of the enzyme (115 kDa). Analysis of the substrate range revealed that the recombinant enzyme catalyzes, in addition to the conversion of some classic peroxidase substrates such as 2,2′-azino-bis(3-ethylthiazoline-6-sulfonate) and substituted phenols like 2,6–dimethoxyphenol, also the decolorization of the anthraquinonic dye Reactive Blue 5. The enzyme also catalyzes bleaching of natural colorants such as β-carotene and annatto. Surprisingly, β-carotene was transformed in the presence and absence of H2O2 by rPsaDyP, however enzyme activity was increased by the addition of H2O2. This indicates that the rPsaDyP has an oxidase function in addition to a peroxidase activity. As a consequence of the high affinity to the characteristic substrate Reactive Blue 5 the rPsaDyP belongs functionally to the dyp-type peroxidase family.
Christiane Lauber; Tatiana Schwarz; Quoc Khanh Nguyen; Patrick Lorenz; Guenter Lochnit; Holger Zorn. Identification, heterologous expression and characterization of a dye-decolorizing peroxidase of Pleurotus sapidus. AMB Express 2017, 7, 1 -15.
AMA StyleChristiane Lauber, Tatiana Schwarz, Quoc Khanh Nguyen, Patrick Lorenz, Guenter Lochnit, Holger Zorn. Identification, heterologous expression and characterization of a dye-decolorizing peroxidase of Pleurotus sapidus. AMB Express. 2017; 7 (1):1-15.
Chicago/Turabian StyleChristiane Lauber; Tatiana Schwarz; Quoc Khanh Nguyen; Patrick Lorenz; Guenter Lochnit; Holger Zorn. 2017. "Identification, heterologous expression and characterization of a dye-decolorizing peroxidase of Pleurotus sapidus." AMB Express 7, no. 1: 1-15.
Phospholipids (PLs), together with hyaluronan and lubricin, are involved in boundary lubrication within human articular joints. Levels of lubricants in synovial fluid (SF) have been found to be associated with the health status of the joint. However, the biosynthesis and release of PLs within human joints remains poorly understood. This study contributes to our understanding of the effects of cytokines on the biosynthesis of PLs using cultured fibroblast-like synoviocytes (FLS) from human osteoarthritic knee joints. Cultured FLS were stimulated with IL-1β, TNFα, IL-6, or inhibitors of cell signaling pathways such as QNZ, SB203580 and SP600125 in the presence of stable isotope-labeled precursors of PLs. Lipids were extracted and quantified using electrospray ionization tandem mass spectrometry (ESI-MS/MS). Our analyses provide for the first time a detailed overview of PL species being synthesized by FLS. IL-1β increased the biosynthesis of both phosphatidylethanolamine (PE) and PE-based plasmalogens. We show here that the NF-κB, p38 MAPK and JNK signaling pathways are all involved in IL-1β-induced PL biosynthesis. IL-6 had no impact on PLs, whereas TNFα increased the biosynthesis of all PL classes. The biosynthesis of various PLs is controlled by IL-1β and TNFα. Our detailed PL species analysis revealed that FLS can partly contribute to the elevated PL levels found in human osteoarthritis (OA) SF. IL-1β in particular stimulates PE and PE-based plasmalogens which can act as cell-protective antioxidants. These results suggest that during OA progression, FLS undergo alterations in their PL composition to adapt to the new diseased environment.
K.D. Sluzalska; G. Liebisch; G. Lochnit; B. Ishaque; H. Hackstein; G. Schmitz; M. Rickert; J. Steinmeyer. Interleukin-1β affects the phospholipid biosynthesis of fibroblast-like synoviocytes from human osteoarthritic knee joints. Osteoarthritis and Cartilage 2017, 25, 1890 -1899.
AMA StyleK.D. Sluzalska, G. Liebisch, G. Lochnit, B. Ishaque, H. Hackstein, G. Schmitz, M. Rickert, J. Steinmeyer. Interleukin-1β affects the phospholipid biosynthesis of fibroblast-like synoviocytes from human osteoarthritic knee joints. Osteoarthritis and Cartilage. 2017; 25 (11):1890-1899.
Chicago/Turabian StyleK.D. Sluzalska; G. Liebisch; G. Lochnit; B. Ishaque; H. Hackstein; G. Schmitz; M. Rickert; J. Steinmeyer. 2017. "Interleukin-1β affects the phospholipid biosynthesis of fibroblast-like synoviocytes from human osteoarthritic knee joints." Osteoarthritis and Cartilage 25, no. 11: 1890-1899.
Macrophages are important in the activation of innate immune responses and in a tissue-specific manner in the maintenance of organ homeostasis. Testicular macrophages (TM), which reside in the testicular interstitial space, comprise the largest leukocyte population in the testes and are assumed to play a relevant function in maintaining testicular immune privilege. Numerous studies have indicated that the interstitial fluid (IF) surrounding the TM has immunosuppressive properties, which may influence the phenotype of TM. However, the identity of the immunosuppressive molecules present in the IF is poorly characterized. We show that the rat testicular IF shifted GM-CSF–induced M1 toward the M2 macrophage phenotype. IF-polarized M2 macrophages mimic the properties of TM, such as increased expression of CD163, high secretion of IL-10, and low secretion of TNF-α. In addition, IF-polarized macrophages display immunoregulatory functions by inducing expansion of immunosuppressive regulatory T cells. We further found that corticosterone was the principal immunosuppressive molecule present in the IF and that the glucocorticoid receptor is needed for induction of the testis-specific phenotype of TM. In addition, TM locally produce small amounts of corticosterone, which suppresses the basal expression of inflammatory genes as a means to render TM refractory to inflammatory stimuli. Taken together, these results suggest that the corticosterone present in the testicular environment shapes the immunosuppressive function and phenotype of TM and that this steroid may play an important role in the establishment and sustenance of the immune privilege of the testis.
Ming Wang; Monika Fijak; Hamid Hossain; Melanie Markmann; Rolf M. Nüsing; Günter Lochnit; Michaela F. Hartmann; Stefan A. Wudy; Lizong Zhang; Huanpeng Gu; Lutz Konrad; Trinad Chakraborty; Andreas Meinhardt; Sudhanshu Bhushan. Characterization of the Micro-Environment of the Testis that Shapes the Phenotype and Function of Testicular Macrophages. The Journal of Immunology 2017, 198, 4327 -4340.
AMA StyleMing Wang, Monika Fijak, Hamid Hossain, Melanie Markmann, Rolf M. Nüsing, Günter Lochnit, Michaela F. Hartmann, Stefan A. Wudy, Lizong Zhang, Huanpeng Gu, Lutz Konrad, Trinad Chakraborty, Andreas Meinhardt, Sudhanshu Bhushan. Characterization of the Micro-Environment of the Testis that Shapes the Phenotype and Function of Testicular Macrophages. The Journal of Immunology. 2017; 198 (11):4327-4340.
Chicago/Turabian StyleMing Wang; Monika Fijak; Hamid Hossain; Melanie Markmann; Rolf M. Nüsing; Günter Lochnit; Michaela F. Hartmann; Stefan A. Wudy; Lizong Zhang; Huanpeng Gu; Lutz Konrad; Trinad Chakraborty; Andreas Meinhardt; Sudhanshu Bhushan. 2017. "Characterization of the Micro-Environment of the Testis that Shapes the Phenotype and Function of Testicular Macrophages." The Journal of Immunology 198, no. 11: 4327-4340.
The most frequent disease of the locomotor system is osteoarthritis (OA), which, as a chronic joint disease, might benefit more from nutrition than acute illnesses. Collagen hydrolysates (CHs) are peptidic mixtures that are often used as nutraceuticals for OA. Three CHs were characterized biochemically and pharmacologically. Our biophysical (MALDI-TOF-MS, NMR, AFM) and fluorescence assays revealed marked differences between CHs of fish (Peptan® F 5000, Peptan® F 2000) and porcine (Mobiforte®) origin with respect to the total number of peptides and common peptides between them. Using a novel dual radiolabeling procedure, no CH modulated collagen biosynthesis in human knee cartilage explants. Peptan® F 2000 enhanced the activities of the aggrecanase ADMATS4 and ADMATS5 in vitro without loss of proteoglycan from cartilage explants; the opposite effect was observed with Mobiforte®. Interleukin (IL)-6, matrix metalloproteinase (MMP)-1, -3 and -13 levels were elevated in explants that were treated with Mobiforte® and Peptan® F 5000, but not with Peptan® F 2000. In conclusion, the heterogeneous peptide composition and disparate pharmacological effects between CHs suggest that the effect of a CH preparation cannot be extrapolated to other formulations. Thus, the declaration of a CH as a safe and effective nutraceutical requires a thorough examination of its pleiotropic effects.
Saskia Schadow; Viktor S. Simons; Guenter Lochnit; Jens Kordelle; Zuzana Gazova; Hans-Christian Siebert; Juergen Steinmeyer. Metabolic Response of Human Osteoarthritic Cartilage to Biochemically Characterized Collagen Hydrolysates. International Journal of Molecular Sciences 2017, 18, 207 .
AMA StyleSaskia Schadow, Viktor S. Simons, Guenter Lochnit, Jens Kordelle, Zuzana Gazova, Hans-Christian Siebert, Juergen Steinmeyer. Metabolic Response of Human Osteoarthritic Cartilage to Biochemically Characterized Collagen Hydrolysates. International Journal of Molecular Sciences. 2017; 18 (1):207.
Chicago/Turabian StyleSaskia Schadow; Viktor S. Simons; Guenter Lochnit; Jens Kordelle; Zuzana Gazova; Hans-Christian Siebert; Juergen Steinmeyer. 2017. "Metabolic Response of Human Osteoarthritic Cartilage to Biochemically Characterized Collagen Hydrolysates." International Journal of Molecular Sciences 18, no. 1: 207.
In multicellular parasites (e.g., nematodes and protozoa), proteins and glycolipids have been found to be decorated with phosphorylcholine (PC). PC can provoke various effects on immune cells leading to an immunomodulation of the host's immune system. This immunomodulation allows long-term persistence but also prevents severe pathology due to downregulation of cellular immune responses. PC-containing antigens have been found to interfere with key proliferative signaling pathways in B and T cells, development of dendritic cells and macrophages, and mast cell degranulation. These effects contribute to the observed modulated cytokine levels and impairment of lymphocyte proliferation. In contrast to glycosphingolipids, little is known about the PC-epitopes of proteins. So far, only a limited number of PC-modified proteins from nematodes have been identified. In this project, PC-substituted proteins and glycolipids in Ascaris suum have been localized by immunohistochemistry in specific tissues of the body wall, intestine, and reproductive tract. Subsequently, we investigated the PCome of A. suum by 2D gel-based proteomics and detection by Western blotting using the PC-specific antibody TEPC-15. By peptide-mass-fingerprint matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), we could identify 59 PC-substituted proteins, which are in involved multiple cellular processes. In addition to membrane proteins like vitellogenin-6, we found proteins with structural (e.g., tubulins) and metabolic (e.g., pyruvate dehydrogenase) functions or which can act in the defense against the host's immune response (e.g., serpins). Initial characterization of the PC-epitopes revealed a predominant linkage of PC to the proteins via N-glycans. Our data form the basis for more detailed investigations of the PC-epitope structures as a prerequisite for comprehensive understanding of the molecular mechanisms of immunomodulation.
Thomas Timm; Julia Grabitzki; Cinar Severcan; Suzan Muratoglu; Lisa Ewald; Yavuz Yilmaz; Guenter Lochnit. The PCome of Ascaris suum as a model system for intestinal nematodes: identification of phosphorylcholine-substituted proteins and first characterization of the PC-epitope structures. Parasitology Research 2016, 115, 1263 -1274.
AMA StyleThomas Timm, Julia Grabitzki, Cinar Severcan, Suzan Muratoglu, Lisa Ewald, Yavuz Yilmaz, Guenter Lochnit. The PCome of Ascaris suum as a model system for intestinal nematodes: identification of phosphorylcholine-substituted proteins and first characterization of the PC-epitope structures. Parasitology Research. 2016; 115 (3):1263-1274.
Chicago/Turabian StyleThomas Timm; Julia Grabitzki; Cinar Severcan; Suzan Muratoglu; Lisa Ewald; Yavuz Yilmaz; Guenter Lochnit. 2016. "The PCome of Ascaris suum as a model system for intestinal nematodes: identification of phosphorylcholine-substituted proteins and first characterization of the PC-epitope structures." Parasitology Research 115, no. 3: 1263-1274.
During first merogony Eimeria bovis forms large macromeronts in endothelial host cells containing >120 000 merozoites I. During multiplication, large amounts of cholesterol are indispensable for the enormous offspring membrane production. Cholesterol auxotrophy was proven for other apicomplexan parasites. Consequently they scavenge cholesterol from their host cell apparently in a parasite-specific manner. We here analyzed the influence of E. bovis infection on endothelial host cell cholesterol metabolism and found considerable differences to other coccidian parasites. Overall, free cholesterol significantly accumulated in E. bovis infected host cells. Furthermore, a striking increase of lipid droplet formation was observed within immature macromeronts. Artificial host cell lipid droplet enrichment significantly improved E. bovis merozoite I production confirming the key role of lipid droplet contents for optimal parasite proliferation. The transcription of several genes being involved in both, cholesterol de novo biosynthesis and low density lipoprotein-(LDL) mediated uptake, was significantly up-regulated at a time in infected cells suggesting a simultaneous exploitation of these two cholesterol acquisition pathways. E. bovis scavenges LDL-derived cholesterol apparently through significantly increased levels of surface LDL receptor abundance and LDL binding to infected cells. Consequently, LDL supplementation significantly improved parasite replication. The up-regulation of the oxidized LDL receptor 1 furthermore identified this scavenger receptor as a key molecule in parasite-triggered LDL uptake. Moreover, cellular cholesterol processing was altered in infected cells as indicated by up-regulation of cholesterol-25-hydroxylase and sterol O-acyltransferase. Overall, these results show that E. bovis considerably exploits the host cell cholesterol metabolism to guarantee its massive intracellular growth and replication.
Penny H. Hamid; Joerg Hirzmann; Katharina Kerner; Gerald Gimpl; Guenter Lochnit; Carlos R. Hermosilla; Anja Taubert. Eimeria bovis infection modulates endothelial host cell cholesterol metabolism for successful replication. Veterinary Research 2015, 46, 1 -17.
AMA StylePenny H. Hamid, Joerg Hirzmann, Katharina Kerner, Gerald Gimpl, Guenter Lochnit, Carlos R. Hermosilla, Anja Taubert. Eimeria bovis infection modulates endothelial host cell cholesterol metabolism for successful replication. Veterinary Research. 2015; 46 (1):1-17.
Chicago/Turabian StylePenny H. Hamid; Joerg Hirzmann; Katharina Kerner; Gerald Gimpl; Guenter Lochnit; Carlos R. Hermosilla; Anja Taubert. 2015. "Eimeria bovis infection modulates endothelial host cell cholesterol metabolism for successful replication." Veterinary Research 46, no. 1: 1-17.
IL-1β is a potent proinflammatory cytokine of the innate immune system that is involved in host defense against infection. However, increased production of IL-1β plays a pathogenic role in various inflammatory diseases, such as rheumatoid arthritis, gout, sepsis, stroke, and transplant rejection. To prevent detrimental collateral damage, IL-1β release is tightly controlled and typically requires two consecutive danger signals. LPS from Gram-negative bacteria is a prototypical first signal inducing pro–IL-1β synthesis, whereas extracellular ATP is a typical second signal sensed by the ATP receptor P2X7 that triggers activation of the NLRP3-containing inflammasome, proteolytic cleavage of pro–IL-1β by caspase-1, and release of mature IL-1β. Mechanisms controlling IL-1β release, even in the presence of both danger signals, are needed to protect from collateral damage and are of therapeutic interest. In this article, we show that acetylcholine, choline, phosphocholine, phosphocholine-modified LPS from Haemophilus influenzae, and phosphocholine-modified protein efficiently inhibit ATP-mediated IL-1β release in human and rat monocytes via nicotinic acetylcholine receptors containing subunits α7, α9, and/or α10. Of note, we identify receptors for phosphocholine-modified macromolecules that are synthesized by microbes and eukaryotic parasites and are well-known modulators of the immune system. Our data suggest that an endogenous anti-inflammatory cholinergic control mechanism effectively controls ATP-mediated release of IL-1β and that the same mechanism is used by symbionts and misused by parasites to evade innate immune responses of the host.
Andreas Hecker; Mira Küllmar; Sigrid Wilker; Katrin Richter; Anna Zakrzewicz; Srebrena Atanasova; Verena Mathes; Thomas Timm; Sabrina Lerner; Jochen Klein; Andreas Kaufmann; Stefan Bauer; Winfried Padberg; Wolfgang Kummer; Sabina Janciauskiene; Martin Fronius; Elke K. H. Schweda; Günter Lochnit; Veronika Grau. Phosphocholine-Modified Macromolecules and Canonical Nicotinic Agonists Inhibit ATP-Induced IL-1β Release. The Journal of Immunology 2015, 195, 2325 -2334.
AMA StyleAndreas Hecker, Mira Küllmar, Sigrid Wilker, Katrin Richter, Anna Zakrzewicz, Srebrena Atanasova, Verena Mathes, Thomas Timm, Sabrina Lerner, Jochen Klein, Andreas Kaufmann, Stefan Bauer, Winfried Padberg, Wolfgang Kummer, Sabina Janciauskiene, Martin Fronius, Elke K. H. Schweda, Günter Lochnit, Veronika Grau. Phosphocholine-Modified Macromolecules and Canonical Nicotinic Agonists Inhibit ATP-Induced IL-1β Release. The Journal of Immunology. 2015; 195 (5):2325-2334.
Chicago/Turabian StyleAndreas Hecker; Mira Küllmar; Sigrid Wilker; Katrin Richter; Anna Zakrzewicz; Srebrena Atanasova; Verena Mathes; Thomas Timm; Sabrina Lerner; Jochen Klein; Andreas Kaufmann; Stefan Bauer; Winfried Padberg; Wolfgang Kummer; Sabina Janciauskiene; Martin Fronius; Elke K. H. Schweda; Günter Lochnit; Veronika Grau. 2015. "Phosphocholine-Modified Macromolecules and Canonical Nicotinic Agonists Inhibit ATP-Induced IL-1β Release." The Journal of Immunology 195, no. 5: 2325-2334.
Phosphorylcholine (PC)-modified biomolecules like lipopolysaccharides, glycosphingolipids, and (glyco)proteins are widespread, highly relevant antigens of parasites, since this small hapten shows potent immunomodulatory capacity, which allows the establishment of long-lasting infections of the host. Especially for PC-modified proteins, structural data is rare because of the zwitterionic nature of the PC substituent, resulting in low sensitivities and unusual but characteristic fragmentation patterns. We have developed a targeted mass spectrometric approach using hybrid triple quadrupole/linear ion trap (QTRAP) mass spectrometry coupled to nanoflow chromatography for the sensitive detection of PC-modified peptides from complex proteolytic digests, and the localization of the PC-modification within the peptide backbone. In a first step, proteolytic digests are screened using precursor ion scanning for the marker ions of choline (m/z 104.1) and phosphorylcholine (m/z 184.1) to establish the presence of PC-modified peptides. Potential PC-modified precursors are then subjected to a second analysis using multiple reaction monitoring (MRM)-triggered product ion spectra for the identification and site localization of the modified peptides. The approach was first established using synthetic PC-modified synthetic peptides and PC-modified model digests. Following the optimization of key parameters, we then successfully applied the method to the detection of PC-peptides in the background of a proteolytic digest of a whole proteome. This methodological invention will greatly facilitate the detection of PC-substituted biomolecules and their structural analysis
Thomas Timm; Christof Lenz; Dietrich Merkel; Christian Sadiffo; Julia Grabitzki; Jochen Klein; Guenter Lochnit. Detection and Site Localization of Phosphorylcholine-Modified Peptides by NanoLC-ESI-MS/MS Using Precursor Ion Scanning and Multiple Reaction Monitoring Experiments. Journal of the American Society for Mass Spectrometry 2014, 26, 460 -471.
AMA StyleThomas Timm, Christof Lenz, Dietrich Merkel, Christian Sadiffo, Julia Grabitzki, Jochen Klein, Guenter Lochnit. Detection and Site Localization of Phosphorylcholine-Modified Peptides by NanoLC-ESI-MS/MS Using Precursor Ion Scanning and Multiple Reaction Monitoring Experiments. Journal of the American Society for Mass Spectrometry. 2014; 26 (3):460-471.
Chicago/Turabian StyleThomas Timm; Christof Lenz; Dietrich Merkel; Christian Sadiffo; Julia Grabitzki; Jochen Klein; Guenter Lochnit. 2014. "Detection and Site Localization of Phosphorylcholine-Modified Peptides by NanoLC-ESI-MS/MS Using Precursor Ion Scanning and Multiple Reaction Monitoring Experiments." Journal of the American Society for Mass Spectrometry 26, no. 3: 460-471.
The structural diversity of human milk oligosaccharides (HMOs) strongly depends on the Lewis (Le) blood group status of the donor which allows a classification of these glycans into three different groups. Starting from 50 μL of human milk, a new high-throughput, standardized, and widely automated mass spectrometric approach has been established which can be used for correlation of HMO structures with the respective Lewis blood groups on the basis of mass profiles of the entire mixture of glycans together with selected fragment ion spectra. For this purpose, the relative abundance of diagnostically relevant compositional species, such as Hex(2)Fuc(2) and Hex(3)HexNAc(1)Fuc(2), as well as the relative intensities of characteristic fragment ions obtained thereof are of key importance. For each Lewis blood group, i.e., Le(a-b+), Le(a+b-), and Le(a-b-), specific mass profile and fragment ion patterns could be thus verified. The described statistically proven classification of the derived glycan patterns may be a valuable tool for analysis and comparison of large sets of milk samples in metabolic studies. Furthermore, the outlined protocol may be used for rapid screening in clinical studies and quality control of milk samples donated to milk banks.
Dennis Blank; Sabine Gebhardt; Kai Maass; Günter Lochnit; Viktoria Dotz; Jennifer Blank; Rudolf Geyer; Clemens Kunz. High-throughput mass finger printing and Lewis blood group assignment of human milk oligosaccharides. Analytical and Bioanalytical Chemistry 2011, 401, 2495 -2510.
AMA StyleDennis Blank, Sabine Gebhardt, Kai Maass, Günter Lochnit, Viktoria Dotz, Jennifer Blank, Rudolf Geyer, Clemens Kunz. High-throughput mass finger printing and Lewis blood group assignment of human milk oligosaccharides. Analytical and Bioanalytical Chemistry. 2011; 401 (8):2495-2510.
Chicago/Turabian StyleDennis Blank; Sabine Gebhardt; Kai Maass; Günter Lochnit; Viktoria Dotz; Jennifer Blank; Rudolf Geyer; Clemens Kunz. 2011. "High-throughput mass finger printing and Lewis blood group assignment of human milk oligosaccharides." Analytical and Bioanalytical Chemistry 401, no. 8: 2495-2510.
A mysterious disease affecting calves, named bovine neonatal pancytopenia (BNP), emerged in 2007 in several European countries. Epidemiological studies revealed a connection between BNP and vaccination with an inactivated vaccine against bovine virus diarrhea (BVD). Alloantibodies reacting with blood leukocytes of calves were detected in serum and colostrum of dams, which have given birth to calves affected by BNP. To understand the linkage between vaccination and the development of alloantibodies, we determined the antigens reacting with these alloantibodies. Immunoprecipitation of surface proteins from bovine leukocytes and kidney cells using sera from dams with a confirmed case of BNP in their gestation history reacted with two dominant protein species of 44 and 12 kDa. These proteins were not detected by sera from dams, free of BVDV and not vaccinated against BVD, and from sera of animals vaccinated with a different inactivated BVD vaccine. The 44 kDa protein was identified by mass spectrometry analysis as MHC I, the other as β-2-microglobulin. The presence of major histocompatibility complex class I (MHC I) in the vaccine was confirmed by Western blot using a MHC I specific monoclonal antibody. A model of BNP pathogenesis is proposed.
Fabian Deutskens; Benjamin Lamp; Christiane M Riedel; Eveline Wentz; Günter Lochnit; Klaus Doll; Heinz-Jürgen Thiel; Till Rümenapf. Vaccine-induced antibodies linked to bovine neonatal pancytopenia (BNP) recognize cattle major histocompatibility complex class I (MHC I). Veterinary Research 2011, 42, 97 -97.
AMA StyleFabian Deutskens, Benjamin Lamp, Christiane M Riedel, Eveline Wentz, Günter Lochnit, Klaus Doll, Heinz-Jürgen Thiel, Till Rümenapf. Vaccine-induced antibodies linked to bovine neonatal pancytopenia (BNP) recognize cattle major histocompatibility complex class I (MHC I). Veterinary Research. 2011; 42 (1):97-97.
Chicago/Turabian StyleFabian Deutskens; Benjamin Lamp; Christiane M Riedel; Eveline Wentz; Günter Lochnit; Klaus Doll; Heinz-Jürgen Thiel; Till Rümenapf. 2011. "Vaccine-induced antibodies linked to bovine neonatal pancytopenia (BNP) recognize cattle major histocompatibility complex class I (MHC I)." Veterinary Research 42, no. 1: 97-97.