68 results on '"Josef Schneider"'
Search Results
2. A Comparison of Six ELISAs in the Detection of Anti-HIV-1 in Zambian Sera1
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Alan F. Fleming, Gerhard Hunsmann, François Guillot, Josef Schneider, Conard Syabula, and Rosemary M. Mwendapole
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Anti hiv 1 ,biology ,business.industry ,virus diseases ,Std clinic ,medicine.disease ,False Negative Result ,Virology ,Acquired immunodeficiency syndrome (AIDS) ,Antigen ,Clinical diagnosis ,Immunology ,medicine ,False positive paradox ,biology.protein ,Antibody ,business - Abstract
Serum samples were collected at the Ndola Central Hospital Cooperbelt Province Zambia from 10 adults with clinical diagnosis of AIDS. Also included were 98 successive patients attending the Sexually Transmitted Diseases (STD) Clinic none with symptoms suggestive of AIDS or ARC. The sera were tested for the presence of antibodies against HIV with 6 ELISAs: 1) H9/HTLV-III (German Primate Center); 2) ELAVIA (Pasteur); 3) Enzygnostic (Behring); 4) Wellcozyme (Wellcome); 5) env-80; and 6) anti-HIV (EIA) Roche. Sera giving positive results with any ELISA were tested further by Western blot (WB) with env-80 antigen and by radio-immunoprecipitation (RIP) with antigen from H9/HTLV-III cells. Results from RIP were the final criteria of the positivity or negativity. 20 of 108 sera were anti-HIV positive confirmed by WB and RIP. 8 of 10 patients suspected of AIDS and 12 of 98 successive unselected patients attending the STD clinic had serologically confirmed infection with HIV-1. All 20 true positive sera were detected correctly by the H9/HTLV-III ELISA but in addition 33 sera gave reactions not confirmed by WB or RIP and were classified as false positives. The ELAVIA gave 4 and Enzygnostic gave 1 false positive result but there were no false positives with Wellcozyme env-80 and anti-HIV (EIA) Roche. All 5 ELISAs other than the H9/HTLV-III failed to detect antibody in the serum of 1 STD patient in whom there was a strong reaction with gp120 alone with RIP; in addition the anti-HIV (EIA) Roche gave 1 other false negative result. The Wellcozyme and env-80 ELISAs gave identical results and both had 95% sensitivity and 100% specificity. In this small series the anti-HIV (EIA) Roche had a slightly lower sensitivity. Enzygnostic and ELAVIA had 95% sensitivity but there were false positives and lower specificity (98 and 95% respectively). The H9/HTLV- III ELISA had 100% sensitivity but only 64% specificity and 37% positive predictive value.
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- 2015
3. Uridine Abrogates Mitochondrial Toxicity Related to Nucleoside Analogue Reverse Transcriptase Inhibitors in Hepg2 Cells
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Nils Venhoff, Eva C Koch, Ulrich A. Walker, Manfred Olschewski, Josef Schneider, and Bernhard Setzer
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Anti-HIV Agents ,Mitochondrion ,Biology ,DNA, Mitochondrial ,Electron Transport Complex IV ,chemistry.chemical_compound ,Tumor Cells, Cultured ,medicine ,Humans ,Pharmacology (medical) ,Uridine ,Pharmacology ,Reverse-transcriptase inhibitor ,Nucleoside analogue ,Zalcitabine ,Membrane Proteins ,Lipid Metabolism ,Nucleotidyltransferase ,medicine.disease ,Molecular biology ,Reverse transcriptase ,Mitochondria ,Isoenzymes ,Mitochondrial toxicity ,Infectious Diseases ,chemistry ,Cyclooxygenase 2 ,Prostaglandin-Endoperoxide Synthases ,Toxicity ,Hepatocytes ,Lactates ,Reverse Transcriptase Inhibitors ,medicine.drug - Abstract
ObjectiveTo assess in vitro if uridine may be suitable to prevent or treat mitochondrial toxicity related to nucleoside analogue reverse transcriptase inhibitors (NRTIs).MethodsHuman HepG2-hepatocytes were exposed to NRTIs with or without uridine for 25 days. Cell growth, lactate production, intracellular lipids, mitochondrial DNA (mtDNA) and the ratio between the respiratory chain components COX II (mtDNA-encoded) and COX IV (nuclear-encoded) were measured.ResultsHepG2 cells exposed to zalcitabine (177 nM) without uridine developed a severe depletion of mtDNA (to 8% of wild-type mtDNA levels), resulting in a decline of cell proliferation and COX II levels, with increased lactate and lipid accumulation. Uridine fully abrogated the adverse effects of zalcitabine on hepatocyte proliferation and normalized lactate synthesis, intracellular lipids and COX II levels by adjusting mtDNA levels to about 65% of NRTI-unexposed control cells. This effect was dose-dependent, with a maximum at 200 μM of uridine. Uridine also rapidly and fully restored cell function when added to cells with established mitochondrial dysfunction (zalcitabine for 15 days) despite continued zalcitabine exposure. Uridine also normalized cell proliferation in HepG2 cells exposed to 36 μM of stavudine and protected HepG2-cells exposed to 7 μM of zidovudine + 8 μM of lamivudine (pyrimidine analogues), but failed to improve cell function or mtDNA in cells exposed to 11.8 or 118 μM of didanosine (a purine analogue).ConclusionsThe pyrimidine precursor uridine may attenuate the mitochondrial toxicity of antiretroviral pyrimidine NRTIs in vitro, and its supplementation may represent a promising strategy in the prevention or treatment of mitochondrial toxicities in HIV-infected patients.
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- 2002
4. Inhibition of HIV-1 in Cell Culture by Synthetic Humate Analogues Derived from Hydroquinone: Mechanism of Inhibition
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Bernhard Seubert, Beate Kary, Josef Schneider, Urs Riede, Christine Männer, Roland Weis, and Albrecht Werner
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HIV Envelope Protein gp120 ,V3 loop ,Biology ,Antiviral Agents ,Virus ,Cell Line ,Cell Fusion ,chemistry.chemical_compound ,Viral entry ,Virology ,Hydroxybenzoates ,Humans ,Humic acid ,Infectivity ,chemistry.chemical_classification ,Syncytium ,Hydroquinone ,Virion ,Peptide Fragments ,Recombinant Proteins ,Hydroquinones ,chemistry ,Biochemistry ,Cell culture ,CD4 Antigens ,HIV-1 ,Oxidation-Reduction ,HeLa Cells - Abstract
Humic acids are natural constituents of soil and ground water and mainly consist of mixtures of polycyclic phenolic compounds. A similar complex of compounds with a mean size of about 1000 Da, designated HS-1500, was synthesized by oxidation of hydroquinone. HS-1500 inhibited HIV-1 infection of MT-2 cells with an IC50 of 50-300 ng/ml and showed a mean cell toxicity of about 600 micrograms/ml. Inhibition of HIV-induced syncytium formation was observed at 10-50 micrograms/ml. Treatment of free and cell-attached HIV with HS-1500 irreversibly reduced its infectivity, whereas the susceptibility of target cells for the virus was not impaired by treatment prior to infection. The HIV envelope protein gp120SU bound to sepharose-coupled HS-1500 and could be eluted by high salt and detergent. HS-1500 interfered with the CD4-induced proteolytic cleavage of the V3 loop of virion gp120SU. Furthermore, binding of V3 loop-specific antibodies was irreversibly inhibited, whereas binding of soluble CD4 to gp120SU on virus and infected cells was not affected. In conclusion, our data suggest, that the synthetic humic acid analogue inhibits the infectivity of HIV particles by interference with a V3 loop-mediated step of virus entry.
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- 1996
5. Infectious Proviral Clones of Chimpanzee Foamy Virus (SFVcpz) Generated by Long PCR Reveal Close Functional Relatedness to Human Foamy Virus
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Josef Schneider, Ottmar Herchenröder, Dieter Neumann-Haefelin, Axel Rethwilm, and Robert Turek
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Transcriptional Activation ,Pan troglodytes ,viruses ,Molecular Sequence Data ,Retroviridae Proteins ,Simian foamy virus ,Biology ,Polymerase Chain Reaction ,Cell Line ,Plasmid ,Proviruses ,Virology ,Virus latency ,medicine ,Animals ,Humans ,Promoter Regions, Genetic ,Gene ,DNA Primers ,Repetitive Sequences, Nucleic Acid ,Base Sequence ,Transfection ,Human foamy virus ,Provirus ,biology.organism_classification ,medicine.disease ,Molecular biology ,Virus Latency ,DNA-Binding Proteins ,genomic DNA ,DNA, Viral ,Trans-Activators ,Spumavirus - Abstract
Infectious proviral clones of simian foamy virus isolated from chimpanzee (SFVcpz) were generated by long PCR. Two overlapping fragments representing the complete provirus were amplified from genomic DNA of infected cells. Four 8.8-kbp amplimers extending from base 1 of the provirus into the env gene and five 4.45-kbp amplimers reaching from env to the end of the 3′-LTR were cloned into pCR II. Subsequently, the proviral fragments were combined in a chessboard manner to generate 20 plasmids containing full-length proviral DNA. Four plasmids produced infectious virus after transfection of susceptible cells. A distinct proviral form bearing a deletion in the transactivator gene joining both exons of a second regulatory gene present in wild-type foamy virus-infected cells started to emerge 48 hr after transfection of BHK cells with infectious SFVcpz DNA. This observation supports a novel hypothesis to explain establishment of foamy virus latency. The transactivator protein Taf of SFVcpz transcomplemented for the homologous protein Bel-1 of the unique human foamy virus isolate (HFV) and Bel-1 exhibited the reciprocal activity, suggesting that HFV could represent a variant of chimpanzee foamy virus.
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- 1995
6. A peptide derived from the highly conserved protein GAPDH is involved in tissue protection by different antifungal strategies and epithelial immunomodulation
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Sabine Nuding, Claudia Borelli, Josef Schneider, Hans Christian Korting, Martin Schaller, Jeanette Wagener, Wolf-Georg Forssmann, Bernhard Hube, Robert Küchler, Jan Wehkamp, Lydia Schild, Hubert Kalbacher, Susann Baxmann, Daniela Mailänder-Sánchez, Christina Braunsdorf, Cornelia Liepke, and Matthias D. Kaeser
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Antifungal Agents ,Aspartic Acid Proteases ,antimicrobial peptide ,Placenta ,Population ,Peptide ,Apoptosis ,Dermatology ,Biology ,Biochemistry ,Epithelium ,Article ,Microbiology ,Cell Line ,Immunomodulation ,03 medical and health sciences ,Pregnancy ,Candida albicans ,Humans ,Secretion ,Interleukin 8 ,education ,Molecular Biology ,Glyceraldehyde 3-phosphate dehydrogenase ,reconstituted human oral epithelium (RHE) ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,education.field_of_study ,030306 microbiology ,GAPDH ,Interleukin-8 ,Candidiasis ,Mouth Mucosa ,Glyceraldehyde-3-Phosphate Dehydrogenases ,Granulocyte-Macrophage Colony-Stimulating Factor ,Cell Biology ,biology.organism_classification ,Corpus albicans ,Peptide Fragments ,Toll-Like Receptor 4 ,chemistry ,Cell culture ,secreted aspartic proteases ,biology.protein ,Female - Abstract
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has an important role not only in glycolysis but also in nonmetabolic processes, including transcription activation and apoptosis. We report the isolation of a human GAPDH (hGAPDH) (2-32) fragment peptide from human placental tissue exhibiting antimicrobial activity. The peptide was internalized by cells of the pathogenic yeast Candida albicans and initiated a rapid apoptotic mechanism, leading to killing of the fungus. Killing was dose-dependent, with 10 μg ml (3.1 μM) and 100 μg ml hGAPDH (2-32) depolarizing 45% and 90% of the fungal cells in a population, respectively. Experimental C. albicans infection induced epithelial hGAPDH (2-32) expression. Addition of the peptide significantly reduced the tissue damage as compared with untreated experimental infection. Secreted aspartic proteinase (Sap) activity of C. albicans was inhibited by the fragment at higher concentrations, with a median effective dose of 160 mg l(-1) (50 μM) for Sap1p and 200 mg l(-1) (63 μM) for Sap2p, whereas Sap3 was not inhibited at all. Interestingly, hGAPDH (2-32) induced significant epithelial IL-8 and GM-CSF secretion and stimulated Toll-like receptor 4 expression at low concentrations independently of the presence of C. albicans, without any toxic mucosal effects. In the future, the combination of different antifungal strategies, e.g., a conventional fungicidal with immunomodulatory effects and the inhibition of fungal virulence factors, might be a promising treatment option.
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- 2012
7. In vivo inhibition of epidermal growth factor receptor autophosphorylation prevents receptor internalization
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Michael Wolff, Franz-Josef Schneider, Birgit Jung, Ralf Heilker, Jens M. Hohlfeld, Michael Chadham Nivens, Kay Tetzlaff, and Publica
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Male ,media_common.quotation_subject ,Biopsy ,education ,Cell Culture Techniques ,Down-Regulation ,Respiratory Mucosa ,Biology ,fluorescence microscopy ,Placebos ,Pulmonary Disease, Chronic Obstructive ,Growth factor receptor ,Double-Blind Method ,Epidermal growth factor ,image analysis ,Humans ,ERBB3 ,Epidermal growth factor receptor ,Phosphorylation ,Receptor ,Internalization ,Protein Kinase Inhibitors ,Cells, Cultured ,media_common ,Autophosphorylation ,Cell Biology ,Cell biology ,internalization ,ErbB Receptors ,Protein Transport ,Cancer research ,biology.protein ,Female ,autophosphorylation ,epidermal growth factor receptor ,Ex vivo - Abstract
The question whether epidermal growth factor (EGF)-induced receptor endocytosis requires the prior autophosphorylation via the EGF receptor (EGFR) kinase domain has been a matter of long-standing debate. In the airway epithelial cell line NCI-H292, the EGFR kinase domain inhibitor BIBW 2948 BS was found to inhibit both autophosphorylation and subsequent internalization of the endogenous EGFR with similar IC₅₀ values. Applying an ex vivo EGFR internalization assay in a clinical study, the in vivo effect of inhalatively administered BIBW 2948 BS was determined directly at the targeted receptor in airway tissues from COPD patients. In these experiments, the in vivo inhibition of the EGFR kinase domain prevented the EGF-induced internalization of EGFR.
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- 2010
8. Comparison of the carbohydrate moieties of recombinant soluble Fc? receptor (sFc? RII/sCD23) expressed inSaccharomyces cerevisiae and Chinese hamster ovary cells. Different O-glycosylation sites are used by yeast and mammalian cells
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Horst Ahorn, Inge Kalsner, Franz-Josef Schneider, Rudolf Geyer, and Ingrid Maurer-Fogy
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Glycosylation ,Molecular Sequence Data ,Saccharomyces cerevisiae ,Carbohydrates ,CHO Cells ,Peptide Mapping ,Biochemistry ,Mass Spectrometry ,law.invention ,chemistry.chemical_compound ,law ,Cricetinae ,Animals ,Humans ,Amino Acid Sequence ,Cloning, Molecular ,Molecular Biology ,chemistry.chemical_classification ,biology ,Receptors, IgE ,Chinese hamster ovary cell ,Cell Biology ,biology.organism_classification ,Molecular biology ,Recombinant Proteins ,Yeast ,Amino acid ,carbohydrates (lipids) ,Carbohydrate Sequence ,chemistry ,biology.protein ,Recombinant DNA ,Electrophoresis, Polyacrylamide Gel ,Antibody ,Glycoprotein - Abstract
Recombinant human soluble low affinity receptor for the Fc portion of IgE (sFc epsilon RII/sCD23) was produced in Saccharomyces cerevisiae or Chinese hamster ovary cells and subjected to carbohydrate analysis. Applied methods included analytical SDS-PAGE, reversed phase HPLC, methylation analysis and sequential degradation with exoglycosidases. The results revealed that sFc epsilon RII derived from Chinese hamster ovary cells is glycosylated exclusively at Ser-147, containing mainly the trisaccharide Sia(alpha 2-3)Gal(beta 1-3)GalNAc, whereas the yeast derived glycoprotein was glycosylated at Ser-167 and contained only alpha-mannosyl residues. It is shown here for the first time that different amino acids of a given protein can be O-glycosylated when expressed in yeast or Chinese hamster ovary cells.
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- 1992
9. Outlines of an 'exploding' network of metabolites generated from the fluoroquinolone enrofloxacin by the brown rot fungus Gloeophyllum striatum
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Josef Schneider, Heinz-Georg Wetzstein, and Wolfgang Karl
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Stereochemistry ,Electrospray ionization ,Metabolite ,Chemical structure ,Applied Microbiology and Biotechnology ,Mass Spectrometry ,Trees ,chemistry.chemical_compound ,Fluoroquinolone Antibiotic ,Enrofloxacin ,medicine ,Organic chemistry ,Chromatography, High Pressure Liquid ,biology ,Molecular mass ,Chemistry ,Isatin ,Basidiomycota ,General Medicine ,biology.organism_classification ,Biodegradation, Environmental ,Culture Media, Conditioned ,Gloeophyllum ,Biotechnology ,medicine.drug ,Fluoroquinolones - Abstract
Degradation of the veterinary fluoroquinolone antibiotic enrofloxacin (EFL) was studied with three strains of Gloeophyllum, basidiomycetous fungi thought to produce extracellular hydroxyl radicals. Metabolites generated in a mineral medium were analyzed by combined high-performance liquid chromatography/high-resolution electrospray ionization mass spectrometry. Their origin was inferred from peak doublets representing 12C and 14C isotopomers detected at a defined proportion. From each exact molecular mass, the molecular formula was derived for which the most probable chemical structure was postulated, using for guidance 18 known EFL metabolites. All supernatants provided similar metabolite patterns, with the most comprehensive consisting of 87 compounds. These metabolites belonged to five families headed by EFL, its oxidatively decarboxylated or defluorinated congeners, an isatin-, and an anthranilic acid-type derivative. Metabolites hydroxylated in the aromatic part suggested the formation of three catechols and two oxidizable ortho-aminophenol-type compounds. After oxidation to the respective ortho-quinones or ortho-quinone imines and oxidative ring cleavage at one of three alternative sites, the formation of various cis,cis-muconic acid-type derivatives is likely, one of which could be detected. Anthranilic acid-type compounds provided two additional sites for ortho-aminophenol formation and aromatic ring cleavage. An “exploding” network of diverse EFL congeners produced by Gloeophyllum suggests the broad utility of our model for studying biodegradation.
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- 2005
10. Human defensins
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Josef Schneider, Angela Unholzer, Hans Christian Korting, Martin Schaller, and Monika Schäfer-Korting
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Chemistry, Pharmaceutical ,Antimicrobial peptides ,medicine.disease_cause ,Microbiology ,Defensins ,Anti-Infective Agents ,Drug Discovery ,medicine ,Animals ,Humans ,Candida albicans ,Escherichia coli ,Genetics (clinical) ,chemistry.chemical_classification ,integumentary system ,biology ,Pseudomonas aeruginosa ,fungi ,biology.organism_classification ,Amino acid ,chemistry ,Staphylococcus aureus ,Streptococcus pyogenes ,Molecular Medicine ,Bacteria ,Antimicrobial Cationic Peptides - Abstract
Antimicrobial peptides are small, cationic, amphiphilic peptides of 12-50 amino acids with microbicidal activity against both bacteria and fungi. The eukaryotic antimicrobial peptides may be divided into four distinct groups according to their structural features: cysteine-free alpha-helices, extended cysteine-free alpha-helices with a predominance of one or two amino acids, loop structures with one intramolecular disulfide bond, and beta-sheet structures which are stabilised by two or three intramolecular disulfide bonds. Mammalian defensins are part of the last-mentioned group. The mammalian defensins can be subdivided into three main classes according to their structural differences: the alpha-defensins, beta-defensins and the recently described theta-defensins. Mammalian alpha-defensins are predominantly found in neutrophils and in small intestinal Paneth cells, whereas mammalian beta-defensins have been isolated from both leukocytes and epithelial cells. Recently, two novel human beta-defensins, human beta-defensin-3 (HBD-3), and human beta-defensin-4 (HBD-4) have been discovered. Similar to HBD-1 and HBD-2, HBD-3 has microbicidal activity towards the Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli) and the yeasts Candida albicans and Malassezia furfur. In addition, HBD-3 kills Gram-positive bacteria such as Streptococcus pyogenes or Staphylococcus aureus, including multi-resistant S. aureus strains, and even vancomycin-resistant Enterococcus faecium. In contrast to HBD-1 and HBD-2, significant expression of HBD-3 has been demonstrated in non-epithelial tissues, such as leukocytes, heart and skeletal muscle. HBD-4 is expressed in certain epithelia and in neutrophils. Its bactericidal activity against P. aeruginosa is stronger than that of the other known beta-defensins. Here we present an overview of human antimicrobial peptides with some emphasis on their antifungal properties.
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- 2004
11. Specific binding of recombinant foamy virus envelope protein to host cells correlates with susceptibility to infection
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Ottmar Herchenröder, Thomas Pietschmann, Josef Schneider, Myra O. McClure, Dieter Moosmayer, Axel Rethwilm, Michael Bock, Roland Weis, and Paul D. Bieniasz
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Pan troglodytes ,viruses ,Recombinant Fusion Proteins ,Gene Expression ,Biology ,Spodoptera ,Antibodies, Viral ,Neutralization ,Divalent ,law.invention ,Cell Line ,Viral envelope ,Viral Envelope Proteins ,law ,Neutralization Tests ,Virology ,Cricetinae ,Animals ,Humans ,Receptor ,Glycoproteins ,chemistry.chemical_classification ,Syncytium ,Ligand (biochemistry) ,Molecular biology ,Fusion protein ,chemistry ,Solubility ,Immunoglobulin G ,Recombinant DNA ,Spumavirus ,Immunoglobulin Heavy Chains - Abstract
The interaction of simian foamy viruses (FVs) with their putative cellular receptor(s) was studied with two types of recombinant envelope protein (Env). Transient expression of full-length Env in BHK-21 cells induced syncytia formation. However, selected stable transfectants fused with naive cells but not with each other. A soluble fusion protein of the Env surface domain with the Fc fragment of a human IgG1 heavy chain (EnvSU–Ig) was produced in the baculovirus expression system, purified to homogeneity, and used for binding and competition analyses. EnvSU–Ig but not unrelated Ig fusion proteins bound to cells specifically. Neutralizing serum blocked binding of EnvSU–Ig and, vice versa, serum-mediated neutralization was abrogated by the chimeric protein. Concomitant reduction of EnvSU–Ig binding and FV susceptibility was seen in Env-expressing target cells. Although EnvSU–Ig did not inhibit FV infection, very likely due to its displacement by multivalent virus–cell interactions, this divalent ligand should help to characterize functionally and to identify the ubiquitous FV receptor.
- Published
- 1999
12. Mammalian glial cells in culture synthesize acetylcholine
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Torsten Reinheimer, Kurt Racké, Holger Klapproth, Franz-Josef Schneider, Ignaz Wessler, and Rudolf Hammer
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Choline O-Acetyltransferase ,chemistry.chemical_compound ,Mice ,medicine ,Animals ,Neurotransmitter ,Cells, Cultured ,Chromatography, High Pressure Liquid ,Pharmacology ,Acetylcholine Bromide ,Microglia ,biology ,General Medicine ,Rat brain ,Choline acetyltransferase ,Enzyme assay ,Acetylcholine ,Cell biology ,Rats ,medicine.anatomical_structure ,chemistry ,Biochemistry ,Animals, Newborn ,Astrocytes ,biology.protein ,Astrocyte ,medicine.drug - Abstract
In the present study we demonstrate that acetylcholine is synthesized by cultured mammalian glial cells identified by cell-type specific markers. Primary cultures of rat brain astrocytes or microglia contained 2.0 and 1.6 pmol acetylcholine/10(6) cells on average respectively. Astrocyte cultures established from neonatal mouse brain contained even more acetylcholine (about 80 pmol acetylcholine/10(6) cells). Primary cultures of rat brain astrocytes showed choline acetyltransferase (ChAT) enzyme activity of 3 nmol/mg protein/h; ChAT activity was blocked by 10 microM bromoacetylcholine. In conclusion, these data demonstrate the synthesis of the "neurotransmitter" acetylcholine in cultured glial cells, a finding which opens a new view upon the role of acetylcholine in mammalian brain.
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- 1997
13. Isolation, cloning, and sequencing of simian foamy viruses from chimpanzees (SFVcpz): high homology to human foamy virus (HFV)
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Krishna K. Murthy, Elaine K. Cobb, Ottmar Herchenröder, Josef Schneider, Paul A. Luciw, Dragutin Loncar, Rolf Renne, and Ayalew Mergia
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Genes, Viral ,Pan troglodytes ,viruses ,Molecular Sequence Data ,Sequence Homology ,Simian foamy virus ,Biology ,Homology (biology) ,Cell Line ,Viral Proteins ,Dogs ,Virology ,Chlorocebus aethiops ,Animals ,Humans ,Amino Acid Sequence ,Cloning, Molecular ,Spumavirus ,Promoter Regions, Genetic ,Gene ,Phylogeny ,Genomic organization ,Repetitive Sequences, Nucleic Acid ,Genetics ,B-Lymphocytes ,Base Sequence ,virus diseases ,biochemical phenomena, metabolism, and nutrition ,Provirus ,Human foamy virus ,biology.organism_classification ,Long terminal repeat ,DNA, Viral - Abstract
Several independent isolates of simian foamy viruses (SFV) were recovered from chimpanzee B-cell lines. One isolate, designated SFVcpz, was molecularly cloned and sequenced. In addition, the genome of SFV type 6 (SFV-6), an independent chimpanzee foamy virus isolate, was partially cloned. The SFVcpz provirus is 13,246 base pairs (bp) long. It is flanked by long terminal repeats (LTRs) and encodes the genes gag, pol, env, the transcriptional transactivator taf, and a second 3′ open reading frame (orf-2). DNA sequences of molecular clones derived from the pol, env, and orf-2 genes of SFV-6 are almost identical to those of SFVcpz. DNA and deduced protein sequences of SFVcpz show high homologies to human foamy virus (HFV), whereas both SFV-1 from a rhesus macaque and SFV-3 from an African green monkey are phylogenetically further distant viruses. Amino acid homologies between corresponding genes of SFVcpz and HFV range between 86% for the taf gene and 95% for the pol gene. Comparisons of taf and pol of SFVcpz with SFV-1 and SFV-3 show 40 and 78% homology, respectively. The SFVcpz LTR consists of 1760 bp and is in the same size range as the LTRs of SFV-1 and -3, but significantly larger than the known HFV LTR. These comparisons reveal that a region approximately 500 bp long is missing in the HFV LTR. We also isolated and sequenced an LTR of a wild-type HFV provirus which aligns with high homology to the SFVcpz LTR without major gaps. Based on sequence comparisons in this report, primate foamy viruses may be arranged into different clusters with SFVcpz and HFV forming one cluster and SFV-1 and SFV-3 as prototypes for two unique clusters.
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- 1994
14. Novel indolocarbazole protein kinase C inhibitors prevent reactivation of HIV-1 in latently infected cells
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Christoph Schächtele, Josef Schneider, Susanne Pätzold, Claus Rudolph, and Dieter Marmé
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Indoles ,Carbazoles ,Antineoplastic Agents ,Indolocarbazole ,Cell Line ,chemistry.chemical_compound ,Virology ,Virus latency ,medicine ,Cytotoxic T cell ,Humans ,Protein kinase C ,Protein Kinase C ,Pharmacology ,biology ,medicine.disease ,Molecular biology ,In vitro ,Virus Latency ,chemistry ,Cell culture ,Enzyme inhibitor ,Tetradecanoylphorbol Acetate ,biology.protein ,HIV-1 ,Virus Activation - Abstract
Suppression of human immunodeficiency virus-1 (HIV-1) reactivation in latently infected cells by protein kinase C (PKC) inhibitors has been described. Based on an initial finding with the indolocarbazole inhibitor Go 6976 we have examined several members of this new class of potent and specific PKC inhibitors with respect to their ability to prevent the PKC-mediated induction of HIV-1 replication in the latently infected U1 cell line. Two of these compounds strongly inhibited not only PMA-induced release of p24-antigen and infectious virus particles into the supernatant (50% inhibition at 0.04-0.35 microM) but also TNF-alpha-mediated HIV-1 reactivation in the same concentration range. Significant lower toxicities compared to Go 6976 were observed for the new compounds, with 50% cytotoxic concentrations at 5.2 microM for Go 7775 and 3.4 microM for Go 7716. This resulted in selectivity indices which were 10-20-times higher compared to the reference compound Go 6976 and were comparable to those of registered anti-AIDS drugs. No anti-HIV-1 activity was observed for a closely related indolocarbazole analogue with no inhibitory activity in the PKC in vitro enzyme assay. This study demonstrates the important role of PKC in reactivation of HIV-1 in latently infected cells and points to the potential of indolocarbazoles to preserve the latent state of HIV-1 infection.
- Published
- 1993
15. A novel proviral clone of HIV-2: Biological and phylogenetic relationship to other primate immunodeficiency viruses
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Wolfgang Lüke, Katja Nieselt, Gerhard Hunsmann, Frank Kirchhoff, Manfred Eigen, Carine Laloux, Klaus Dieter Jentsch, Andreas W. Stuke, Christiane Stahl-Hennig, Josef Schneider, and Barbara Bachmann
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Primates ,Genes, Viral ,Molecular Sequence Data ,Restriction Mapping ,Clone (cell biology) ,medicine.disease_cause ,Macaque ,Polymerase Chain Reaction ,Virus ,law.invention ,Cell Line ,Proviruses ,Viral Envelope Proteins ,Phylogenetics ,law ,Virology ,biology.animal ,Sequence Homology, Nucleic Acid ,medicine ,Animals ,Humans ,Cloning, Molecular ,Gene ,Polymerase chain reaction ,Phylogeny ,Genetics ,biology ,Phylogenetic tree ,Base Sequence ,virus diseases ,Simian immunodeficiency virus ,Macaca mulatta ,Retroviridae ,HIV-2 - Abstract
Infectious molecular clones of the human immunodeficiency virus type 2 (HIV-2) will be valuable tools for the study of regulatory gene functions and the development of an animal model for the human acquired immunodeficiency syndrome (AIDS). To this end, we have cloned and sequenced a novel HIV-2 isolate, HIV-2BEN. One clone, designated MK6, is infectious for various human T-cell lines and for human and macaque peripheral blood lymphocytes (PBL), allowing molecular studies of HIV-2 infection and replication. Since MK6 is highly cytopathic in MT-2 and Molt-4 clone 8 cells, antiviral agents and neutralizing sera may be tested. Cluster analysis of HIV-1, HIV-2, and simian immunodeficiency virus (SIV) env and gag genes revealed that HIV-2BEN yielded the earliest node of phylogenetic divergence for all reported HIV-2 sequences. Noise analysis showed that, with the current data, no specification of any branching order can be made among the four groups of primate lentiviruses, HIV-1, HIV-2/SIVSMM/MAC, SIVAGM, and SIVMND.
- Published
- 1990
16. Immunoprevention of Friend Leukaemia Virus-induced Erythroleukaemia by Vaccination with Aggregated gp70
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Christoph Kleiser, Gerhard Hunsmann, Josef Schneider, and H Bayer
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Friend leukemia ,viruses ,Antibodies, Viral ,Virus ,Microbiology ,Epitopes ,Mice ,Viral Envelope Proteins ,Viral envelope ,hemic and lymphatic diseases ,Virology ,Murine leukemia virus ,Animals ,Leukemia, Experimental ,biology ,Viral Core Proteins ,Viral Vaccine ,Viral Vaccines ,biology.organism_classification ,Friend murine leukemia virus ,carbohydrates (lipids) ,Vaccination ,Glutaral ,Hemocyanins ,biology.protein ,Female ,Leukemia, Erythroblastic, Acute ,Antibody ,Keyhole limpet hemocyanin - Abstract
Vaccines prepared from Friend leukaemia virus envelope and core polypeptides were compared for their efficiency in preventing erythroleukaemia in mice. High doses (100 micrograms) of gp85, the micellar complex of the envelope polypeptides gp70 and p15E, completely protected STU mice. The same dose of purified gp70 still protected about 80% of the animals, while p15E did not affect the cumulative mortality. The internal viral polypeptide p30 was ineffective. Serological examination indicated that immunity against death from leukaemia was mediated by specific antibodies. These leukaemia-preventing antibodies were predominantly induced by immunization with the gp70 env gene product, since p15E showed only minor protection. Glycoprotein gp70, however, was more effective when given as the gp85 micellar complex. An even more potent vaccine was obtained when gp70 was coupled to keyhole limpet haemocyanin (KLH) by glutaraldehyde. Ten micrograms gp70 coupled to KLH was enough to save more than 90% of Friend leukaemia virus-infected mice from erythroleukaemia. KLH may also be a suitable experimental carrier for subunits of gp70 or synthetic oligopeptides for viral vaccines.
- Published
- 1986
17. A glycopolypeptide (gp 100) is the main antigen detected by HTLV-III antisera
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Gerhard Hunsmann, Ulrich Bienzle, Josef Schneider, and H Bayer
- Subjects
Male ,Microbiology (medical) ,Antigenicity ,viruses ,Immunology ,Antibodies, Viral ,Hemophilia A ,Deltaretrovirus ,Virus ,Viral Proteins ,Viral Envelope Proteins ,Antigen ,Viral envelope ,Antibody Specificity ,Humans ,Immunology and Allergy ,Glycoproteins ,chemistry.chemical_classification ,Antiserum ,biology ,Homosexuality ,General Medicine ,Virology ,Molecular biology ,Molecular Weight ,Titer ,chemistry ,biology.protein ,Antibody ,Glycoprotein - Abstract
Sera from homosexuals and hemophiliacs in Germany were examined for antibodies to human T-lymphotropic retrovirus type III (HTLV-III) by an enzyme-linked immunosorbent assay (ELISA) against purified virus. ELISA positive sera were used to search by immunoprecipitation for HTLV-III related antigens in a persistently infected human T-cell line. A glycopolypeptide with a mol. wt. of 100 000 was regularly recognized by all positive sera. In analogy to glycosylation and high antigenicity of envelope polypeptides of other mammalian retroviruses, gp 100 seemed to be related to the env gen. Two polypeptides with mol. wts. of 24 000 and 22 000 probably representing viral core polypeptides were additionally detected by sera with high ELISA titers.
- Published
- 1985
18. Antibodies to adult T-cell leukemia virus (ATLV/HTLV-I) in AIDS patients and people at risk of AIDS in Germany
- Author
-
M. Dietrich, K. Schimpf, Herbert Schmitz, Ulrich Bienzle, Peter Kern, H. Kabisch, Gerhard Hunsmann, H. Berthold, Klaus Ritter, H Bayer, and Josef Schneider
- Subjects
Adult ,Male ,Microbiology (medical) ,medicine.medical_specialty ,Adolescent ,medicine.medical_treatment ,Immunology ,T-cell leukemia ,Antibodies, Viral ,Hemophilia A ,Deltaretrovirus ,Virus ,03 medical and health sciences ,0302 clinical medicine ,Medical microbiology ,Acquired immunodeficiency syndrome (AIDS) ,medicine ,Humans ,Immunology and Allergy ,Aged ,030304 developmental biology ,Acquired Immunodeficiency Syndrome ,0303 health sciences ,biology ,business.industry ,Germany, West ,Transfusion Reaction ,Homosexuality ,General Medicine ,Middle Aged ,medicine.disease ,Virology ,3. Good health ,Leukemia ,030220 oncology & carcinogenesis ,biology.protein ,Viral disease ,Hemodialysis ,Antibody ,business - Abstract
A total of 2048 serum samples from Germany were examined for antibodies to adult T-cell leukemia virus (ATLV) structural polypeptides with an enzyme-linked immuno sorbent assay (ELISA) and confirmative immuno precipitation. The origin of the sera samples was: 850 samples taken for virological or protozoal diagnosis; 626 samples from male homosexuals, about 20% of whom had lymphadenopathy syndrome; 164 from hemophiliacs; 184 were from multiple transfused, mostly dialysis patients; 9 from intravenous drug abusers; 182 from suspected cases of acquired immuno deficiency syndrome (AIDS) and 33 from AIDS-patients. In none of these sera did we detect antibodies to ATLV, except in the serum of one patient who had been on hemodialysis for over 11 years. Obviously infection with ATLV or a serologically related agent is very rare in our country and an association with AIDS could not be observed.
- Published
- 1985
19. Antibodies to ATLV/HTLV-1 in Africa
- Author
-
H Bayer, D. W. Büttner, M. Dietrich, AlanF. Fleming, Peter Kern, A. M. Goudeau, Gerhard Hunsmann, G. Kulkarni, Josef Schneider, and Herbert Schmitz
- Subjects
Adult ,Microbiology (medical) ,medicine.medical_specialty ,Adolescent ,Immunology ,Antibodies, Viral ,Deltaretrovirus ,Asymptomatic ,Virus ,Medical microbiology ,parasitic diseases ,Epidemiology ,medicine ,Humans ,Immunology and Allergy ,Child ,Aged ,biology ,business.industry ,Age Factors ,Infant, Newborn ,Infant ,Endemic area ,General Medicine ,Middle Aged ,Virology ,Leukemia, Lymphoid ,Titer ,Oncovirinae ,Child, Preschool ,Africa ,Carrier State ,biology.protein ,Antibody ,medicine.symptom ,Epidemiologic Methods ,business ,Retroviridae Infections - Abstract
Almost 4000 sera from seven African states were examined for antibodies to ATLV/HTLV-1. Between 1% and 8% of healthy people from sub-Saharan Africa have such antibodies. The highest frequency was observed in Gabon. There were considerable variations between villages. The percentage of seropositives and the mean titre increased with age. Our findings suggest that the African continent is the largest endemic area for ATLV.
- Published
- 1984
20. Separation procedure and sugar composition of oligosaccharides in the surface glycoprotein of friend murine leukemia virus
- Author
-
Gerhard Hunsmann, Stephan Stirm, Josef Schneider, Hildegard Geyer, and Rudolf Geyer
- Subjects
chemistry.chemical_classification ,Chromatography ,biology ,Surface Properties ,Glycopeptides ,Biophysics ,Oligosaccharides ,Mannose ,Oligosaccharide ,Biochemistry ,Chromatography, Affinity ,Fucose ,Friend murine leukemia virus ,Sialic acid ,chemistry.chemical_compound ,chemistry ,Glucosamine ,Concanavalin A ,Galactose ,biology.protein ,Sugar ,Molecular Biology ,Glycoproteins - Abstract
The sugar composition of the surface glycoprotein from Friend murine leukemia virus was determined by gas-liquid chromatography of the alditol acetates and by the thiobarbituric acid method, respectively. N-Acetylglucosamine, mannose, galactose, sialic acid and fucose were found in a molar ratio around 15.2:11.6:7.4:3.3:1.0. Ten oligosaccharide fractions were obtained from glycoprotein preparations by a suitable sequence of degradation (with pronase, endo-beta-N-acetylglucosaminidase H, neuraminidase, and by hydrazinolysis) and separation procedures (concanavalin A-affinity chromatography and gel filtration). The qualitative sugar composition of these fractions was analyzed by in vivo labelling with D-[6-(3)H]glucosamine, D-[2-(3)H]mannose, D-[6-(3)H]galactose, or L-[6-(3)H]fucose, and their molecular weights were estimated from the gel elution volumina. Four fractions of N-glycosidically linked oligosaccharides of the oligomannosidic ('high mannose') type oligomannosidic7-oligomannosidic10, about seven to ten sugar residues), two of the mixed (M11 and M12), and four of the N-acetyllactosaminic ('complex') type (N-acetyllactosaminic9, probably nine sugar residues; N-acetyllactosaminica-N-acetyllactosaminic c, size unknown) were thus identified.
- Published
- 1982
21. Simian lentiviruses — the SIV group
- Author
-
Josef Schneider and Gerhard Hunsmann
- Subjects
Primates ,Serotype ,Genes, Viral ,Immunology ,HIV ,Biology ,Simian ,biology.organism_classification ,Virology ,Retroviridae ,Infectious Diseases ,Cytopathogenic Effect, Viral ,Group (periodic table) ,Animals ,Humans ,Immunology and Allergy ,Serotyping ,Gene ,Retroviridae Infections - Published
- 1988
22. Structural and immunological characterization of Friend murine leukaemia virus glycopolypeptide using synthetic oligopeptides
- Author
-
W Gruber, H Bayer, Gerhard Hunsmann, and Josef Schneider
- Subjects
Protein Conformation ,Antibodies, Viral ,General Biochemistry, Genetics and Molecular Biology ,Epitope ,Mice ,Viral Proteins ,Antibody Specificity ,hemic and lymphatic diseases ,Animals ,Amino Acid Sequence ,Bovine serum albumin ,Molecular Biology ,Peptide sequence ,Glycoproteins ,Antiserum ,chemistry.chemical_classification ,Oligopeptide ,General Immunology and Microbiology ,biology ,General Neuroscience ,Molecular biology ,Friend murine leukemia virus ,Amino acid ,Biochemistry ,chemistry ,biology.protein ,Glycoprotein ,Oligopeptides ,Keyhole limpet hemocyanin ,Research Article - Abstract
Using terminal position, hydrophilicity, predicted reverse turns and type specificity as criteria, five oligopeptides were selected for synthesis from the amino acid sequence of the envelope glycopolypeptide gp70 of Friend murine leukaemia virus. These peptides corresponded to the amino acids 6-12 (pep1), 124-131 (pep2), 256-262 (pep3), 283-290 (pep4) and 434-441 (pep5). After coupling to carriers, bovine serum albumin or keyhole limpet hemocyanin, antisera were prepared in rabbits. All of the five oligopeptides were immunogenic and pep1, pep2, pep4 and pep5 were able to elicit antibodies to the native glycopolypeptide. These sequence-specific antisera distinguished between glycoproteins of different leukaemia viruses. At least three of the selected peptides, the type-specific oligopeptides pep3, pep4 and pep5, were found to be natural epitopes of gp70.
- Published
- 1984
23. Major oligosaccharides in the glycoprotein of Friend murine leukemia virus: structure elucidation by one- and two-dimensional proton nuclear magnetic resonance and methylation analysis
- Author
-
Stephan Stirm, Josef Schneider, Gerhard Hunsmann, Rudolf Geyer, Janusz Dabrowski, Hildegard Geyer, and Ursula Dabrowski
- Subjects
Magnetic Resonance Spectroscopy ,Glycoside Hydrolases ,Size-exclusion chromatography ,Oligosaccharides ,Methylation ,Biochemistry ,Chromatography, Affinity ,Viral Proteins ,Affinity chromatography ,Acetylglucosaminidase ,Murine leukemia virus ,Carbohydrate Conformation ,Glycoproteins ,chemistry.chemical_classification ,biology ,Hydrolysis ,Nuclear magnetic resonance spectroscopy ,Hydrogen-Ion Concentration ,Oligosaccharide ,biology.organism_classification ,Friend murine leukemia virus ,Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase ,chemistry ,Concanavalin A ,Chromatography, Gel ,Proton NMR ,biology.protein ,Glycoprotein - Abstract
The highly microheterogeneous, N-glycosidically linked oligosaccharides in the glycoproteins of Friend murine leukemia virus (as produced by Eveline cells) were liberated with endo-beta-N-acetylglucosaminidase H and by alkaline hydrolysis. They were fractionated (as desialylated oligosaccharitols) by gel filtration and by concanavalin A affinity chromatography, and the major fractions were analyzed by methylation-gas chromatography-mass spectrometry, by digestion with exoglycosidases, and, especially, by one- and two-dimensional proton nuclear magnetic resonance spectroscopy. Guidelines for qualitative and quantitative analysis of complex oligosaccharide mixtures by NMR were worked out and the results compared with those obtained by methylation analysis. It was found that these major fractions consist of bi-, tri-, and tetraantennary oligosaccharitols of the "complex" type (comprising a minority of species with N-acetyllactosamine repeating units), which are, in part, substituted by nonreducing terminal Gal alpha (1----3) and/or bisecting GlcNAc beta (1----4) residues.
- Published
- 1984
24. Detection of serum antibodies to adult T-cell leukemia virus in non-human primates and in people from Africa
- Author
-
Jakob Schmitt, Naoki Yamamoto, Gerhard Hunsmann, and Josef Schneider
- Subjects
Cancer Research ,biology ,T-Lymphocytes ,T-cell leukemia ,Cercopithecus ,Antibodies, Viral ,medicine.disease ,Kenya ,Virology ,Virus ,Leukemia ,Retroviridae ,Oncology ,German population ,Germany ,biology.animal ,Chlorocebus aethiops ,Immunology ,medicine ,biology.protein ,Animals ,Humans ,Primate ,African Green Monkey ,Antibody - Abstract
The distribution of serum antibodies to adult T-cell leukemia virus (ATLV) was examined as a marker for virus infection among non-human primates as well as people from Africa and Germany. The virus is present in Africa in certain primate species including man. Altogether, 468 sera from 27 monkey species were examined. Only African green monkeys, less frequently also chimpanzees and crab-eating monkeys, were found to be infected. About 1-2% of people from Kenya have antibodies, while ATLV-antibodies may be present in well below 0.1% of the German population.
- Published
- 1983
25. Surface expression of murine leukemia virus structural polypeptides on host cell and the virion
- Author
-
Josef Schneider and Gerhard Hunsmann
- Subjects
Cytotoxicity, Immunologic ,Peptide Biosynthesis ,Cancer Research ,viruses ,Viral transformation ,Antibodies, Viral ,Virus ,Epitope ,Cell Line ,Epitopes ,hemic and lymphatic diseases ,Murine leukemia virus ,Antigens, Viral ,Polyacrylamide gel electrophoresis ,Antiserum ,biology ,Friend virus ,biology.organism_classification ,Molecular biology ,Friend murine leukemia virus ,Leukemia Virus, Murine ,Cell Transformation, Neoplastic ,Oncology ,Antigens, Surface ,biology.protein ,Antibody ,Peptides - Abstract
The expression of murine leukemia virus structural polypeptides on the surface of cells producing exogenous Friend leukemia virus, endogenous ecotropic AKR and xenotropic BALB/c virus was investigated. Antisera to Friend virus gp71, p30, p15E, p12 and p10 were employed in a complement-dependent chromium release assay and to immunoprecipitate lactoperoxidase iodinated surface polypeptides prior to analysis in polyacrylamide gel electrophoresis. With the latter technique gag-gene encoded proteins and their precursors were not discovered on the viral and cellular surface membranes. Only env-gene encoded polypeptides gp85, gp71, and p15E were detectable. p15E is embedded into the lipid membrane. gp85 is formed by disulfide linkage of p15E to surface-exposed gp71. The ratio of gp71 to gp85 is variable and apparently determined by the host cell. Antibodies of strong cytotoxicity are those against type- and group-specific epitopes of gp71 as well as type-specific epitopes of p12.
- Published
- 1978
26. Serological and Structural Comparison of Immunodeficiency Viruses from Man, African Green Monkey, Rhesus Monkey and Sooty Mangabey
- Author
-
Ohta Y, Gerhard Hunsmann, Jurkiewicz E, Masanori Hayami, Josef Schneider, and Herbert Schmitz
- Subjects
viruses ,animal diseases ,Cercopithecus ,Cross Reactions ,Simian ,Deltaretrovirus ,Cercopithecus aethiops ,General Biochemistry, Genetics and Molecular Biology ,Serology ,Species Specificity ,Chlorocebus aethiops ,medicine ,Humans ,Serotyping ,Immunodeficiency ,chemistry.chemical_classification ,biology ,HIV ,virus diseases ,Cercopithecidae ,Haplorhini ,biology.organism_classification ,medicine.disease ,Macaca mulatta ,Virology ,Rhesus macaque ,chemistry ,Sooty mangabey ,Macaca ,African Green Monkey ,Glycoprotein - Abstract
We have studied the serological relationship among the human immunodeficiency virus type 1 (HIV-1), and three simian immunodeficiency viruses (SIV). SIVagm was isolated from African green monkeys (Cercopithecus aethiops), and compared with the previously described isolates of SIVmac from a rhesus macaque (Macaca mulatta) and SIVsm from a sooty mangabey (Cercocebus atys). With respect to the glycoproteins, the simian viruses represent a subgroup apparently different from HIV . To classify HIV and SIV isolates further, we compared tryptic peptide maps of the core polypeptides p18 and p24 of HIV-2, three HIV-1 and five SIV isolates. Each peptide map was distinguishable, and differences are most prominent between the HIV-1 group and the SIVmac/SIVsm group. HIV-2 is very similar to SIVmac and SIVsm. The three SIVagm isolates form a more heterogeneous group. The p24s of all SIVagms are more similar to the p24s of HIV-1, but with respect to p18, one isolate is similar to HIV-1, while the two others are more related to SIVmac, SIVsm, and HIV-2.
- Published
- 1988
27. Highly efficient neutralization of HIV with recombinant CD4-immunoglobulin molecules
- Author
-
Klaus Karjalainen, Hansruedi Kiefer, Josef Schneider, and André Traunecker
- Subjects
Virus genetics ,Complement Activating Enzymes ,Macromolecular Substances ,Immunoglobulin gamma-Chains ,Recombinant Fusion Proteins ,Immunoglobulin Variable Region ,Retroviridae Proteins ,Immunoglobulins ,Receptors, Fc ,HIV Envelope Protein gp120 ,Immunoglobulin light chain ,Antiviral Agents ,law.invention ,Mice ,Receptors, HIV ,Complement C1 ,law ,Animals ,Complement C1q ,Acquired Immunodeficiency Syndrome ,Multidisciplinary ,biology ,Immunoglobulin mu-Chains ,Molecular biology ,Recombinant Proteins ,Cell biology ,Immunoglobulin M ,Immunoglobulin G ,HIV-1 ,biology.protein ,Recombinant DNA ,Receptors, Virus ,Immunoglobulin Constant Region ,Antibody ,Immunoglobulin Constant Regions ,Plasmids - Abstract
The human immunodeficiency virus type 1 (HIV-1) exploits the cell surface CD4 molecule to initiate the infection which can lead, eventually, to acquired immunodeficiency syndrome (AIDS). The HIV-1 envelope protein, gp120, interacts specifically with CD4 and soluble CD4 molecules have been shown to inhibit HIV infectivity in vitro. Effective inhibition in vivo may, however, require more potent reagents. We describe here the generation of molecules which combine the specificity of CD4 and the effector functions of different immunoglobulin subclasses. Replacing the VH and CH1 domains of either mouse gamma 2a or mu heavy chains with the first two N-terminal domains of CD4 results in molecules that are secreted in the absence of any immunoglobulin light chains. We find that the pentameric CD4-IgM chimaera is at least 1,000-fold more active than its dimeric CD4-IgG counterpart in syncytium inhibition assays and that effector functions, such as the binding of Fc receptors and the first component of the complement cascade (Clq), are retained. Similar chimaeric molecules, combining CD4 with human IgG were recently described by Capon et al., but these included the CH1 domain and did not bind Clq. Deletion of the CH1 domain may allow the association and secretion of heavy chains in the absence of light chains, and we suggest that the basic design of our constructs may be generally and usefully applied.
- Published
- 1989
28. Analysis of the phase variation in lambda reduced immunity lysogens
- Author
-
Josef Schneider and Hans B. Strack
- Subjects
DNA Replication ,DNA, Bacterial ,viruses ,Mutant ,Genome ,Coliphages ,Bacteriophage ,Lysogen ,Immunity ,Lysogenic cycle ,Genetics ,Escherichia coli ,Molecular Biology ,Lysogeny ,Phase variation ,biology ,Transition (genetics) ,Nucleic Acid Hybridization ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,Molecular biology ,DNA, Viral ,Mutation ,bacteria ,Cell Division - Abstract
Two distinct phases characterized by different levels of immunity that appear in some E. coli strains lysogenic for reduced immunity mutants of bacteriophage lambda are identified as single and tandem double lysogens respectively on the basis of DNA-DNA hybridization experiments and the requirement of the phage xis function for the transition from a single to a double, and of the host recA function for the transition from a double to a single lysogen (in a xis- condition). Rim lysogens with a further increase in immunity, containing some 5 copies of the lambda genome per host genome, have also been observed. It is argued that the different levels of immunity are a direct reflection of the CI gene dosage effect. An unexplained finding is that rim single lysogens yield double lysogens with a frequency of near 1% per generation, whereas cured cells fail to appear even at a frequency 100 times lower.
- Published
- 1977
29. Binding of adult T-cell leukemia virus to various hematopoietic cells
- Author
-
Tadafumi S. Tochikura, Naoki Yamamoto, Josef Schneider, Yoshio Koyanagi, Toru Chosa, and Yorio Hinuma
- Subjects
Cancer Research ,viruses ,Hematopoietic System ,T-cell leukemia ,Fluorescent Antibody Technique ,Biology ,Immunofluorescence ,Deltaretrovirus ,Virus ,Cell Line ,Viral Proteins ,Retrovirus ,Antigens, Neoplasm ,Blocking antibody ,medicine ,Humans ,Glycoproteins ,medicine.diagnostic_test ,medicine.disease ,biology.organism_classification ,Virology ,Molecular biology ,Haematopoiesis ,Leukemia ,Oncology ,Receptors, Virus - Abstract
A newly found human retrovirus, adult T-cell leukemia virus (ATLV) was shown by means of membrane immunofluorescence to bind to various hematopoietic cells including T-, B- and non-T, non-B-cell lines. Partially purified viral gp46 from culture fluids of ATL virus producer lines also bound efficiently to an ATLV-negative T-cell line, CCRF-CEM cells. When the viruses were pre-incubated with anti-ATLV-positive human sera, ATLV binding to the cells was clearly inhibited but not by pre-incubation with anti-ATLV-negative sera. These data suggest that: (1) ATLV binds not only to T-cells but also to multiple types of cells of hematopoietic origin; (2) anti-ATLV antibody-positive human sera have the blocking antibody for the binding of ATLV to lymphoid cells.
- Published
- 1984
30. Efficient replication of HTLV-III and STLV-IIImac in human Jurkat cells
- Author
-
Josef Schneider, K. D. Jentsch, Gerhard Hunsmann, and I Wendler
- Subjects
Microbiology (medical) ,viruses ,T-Lymphocytes ,Immunology ,Biology ,Virus Replication ,Jurkat cells ,Virus ,Cell Line ,Viral Proteins ,Antigen ,Antibody Specificity ,Immunology and Allergy ,Animals ,Humans ,Antigens, Viral ,Acquired Immunodeficiency Syndrome ,HIV ,RNA-Directed DNA Polymerase ,General Medicine ,Virology ,Macaca mulatta ,Leukemia, Lymphoid ,Cell culture ,biology.protein ,Antibody ,Htlv iii - Abstract
High producer cell lines were established by infecting Jurkat cells with either HTLV-IIIB from H9 cells or with STLV-IIImac from HUT-78 cells. The Jurkat cells produced both viruses at least 10 times more efficiently then the original cell lines; the pelleted virus from Jurkat cells was also less contaminated with non-virion proteins. Accordingly, a higher specificity was attained in an ELISA for antibodies to HTLV-III if the antigenic activity was derived from the virus from Jarkat cells, as opposed to that from H9 cells.
- Published
- 1987
31. Synthesis and expression of MHC class II molecules in the absence of attached invariant chains by recombinant-interferon-gamma-activated bone-marrow-derived macrophages
- Author
-
Wolfgang Ballhausen, Konrad Reske, Peter Steinlein, Franz-Josef Schneider, Wolfgang Henkes, and Beate Opel
- Subjects
Immunology ,Bone Marrow Cells ,law.invention ,Interferon-gamma ,Mice ,law ,Immunology and Allergy ,Animals ,Northern blot ,RNA, Messenger ,Gel electrophoresis ,Messenger RNA ,MHC class II ,Mice, Inbred C3H ,Polymorphism, Genetic ,biology ,Isoelectric focusing ,Macrophages ,Histocompatibility Antigens Class II ,DNA ,Macrophage Activation ,Molecular biology ,In vitro ,Recombinant Proteins ,Gene Expression Regulation ,Recombinant DNA ,biology.protein ,Intracellular - Abstract
Pure populations of in vitro propagated bone marrow-derived macrophages are constitutively Ia negative. Co-culturing of these cells with recombinant interferon-gamma (rIFN-gamma) resulted in the appearance of high amounts of Ia antigens at the cell surface of essentially all cells. The continuous presence of the stimulus was a prerequisite for sustained Ia expression because removal of the stimulus resulted in rapid decline of surface Ia. Two-dimensional (2D) gel analysis (1D isoelectric focusing, 2D sodium dodecyl sulfate-polyacrylamide gel electrophoresis) of class II molecules synthesized by rIFN-gamma-stimulated bone marrow macrophages (BMM phi) revealed that, in contrast to class II complexes hitherto described, BMM phi-derived I-A and I-E subregion-encoded subunits are synthesized without invariant chains. The invariant chain-deficient alpha,beta heterodimers are expressed at the cell surface in high proportions demonstrating that their correct assembly and transport to the cell surface is accomplished in the absence of invariant chains. The lack of invariant chains appears not to be due to a failure of rIFN-gamma to induce transcription of the gamma-chain gene because rIFN-gamma-induced, in contrast to uninduced, BMM phi accumulate high levels of invariant chain-specific transcripts as evidenced by Northern blot analysis. These findings suggest that translation of gamma-chain-specific mRNA is blocked in BMM phi for as yet unknown reasons. Alternatively, newly synthesized gamma chains might have escaped their regular intracellular maturation pathway as a result of unidentified modifications mediated by altered post-translational processing mechanisms.
- Published
- 1987
32. Virosomes constructed from lipid and purified Friend leukaemia virus glycoprotein
- Author
-
Josef Schneider, Heinz Falk, and Gerhard Hunsmann
- Subjects
animal structures ,Hemagglutination ,viruses ,medicine.medical_treatment ,Virus ,Viral Proteins ,Viral envelope ,Viral Envelope Proteins ,hemic and lymphatic diseases ,Virology ,medicine ,chemistry.chemical_classification ,Liposome ,Protease ,biology ,Friend murine leukemia virus ,Microscopy, Electron ,Membrane ,Biochemistry ,chemistry ,Concanavalin A ,embryonic structures ,Liposomes ,biology.protein ,Glycoprotein ,Dialysis - Abstract
Summary Liposomes were loaded by a dialysis technique with purified envelope glycopolypeptide, gp85, of the Friend murine leukaemia virus (F-MuLV). The gp85 liposomes prepared from cellular lipid had a buoyant density of 1.05 g/ml and an apparent diameter of 50 to 300 nm. The gp85 was not simply entrapped by liposomes nor adsorbed non-specifically to their outer surface. Experiments with radioactively labelled protein, electron microscopic examinations, protease treatment and concanavalin A binding showed that gp85 is anchored in the liposomal membrane and orientated asymmetrically as in the virus envelope. Moreover, gp85-covered liposomes displayed some functions of the intact F-MuLV envelope, such as absorption of antibodies to gp70 and haemagglutination after enzyme treatment. To some extent, these lipid vesicles appeared to be reconstituted F-MuLV envelopes and thus, by analogy to other systems, were named retrovirosomes.
- Published
- 1983
33. Experimental infection of cynomolgus monkeys with a human retrovirus, adult T-cell leukemia virus
- Author
-
Gerhard Hunsmann, Naoki Yamamoto, Masanori Hayami, Morihisa Okada, Atsumi Komuro, Josef Schneider, and Yorio Hinuma
- Subjects
Microbiology (medical) ,Male ,Time Factors ,viruses ,Lymphocyte ,Immunology ,T-cell leukemia ,Antibodies, Viral ,Deltaretrovirus ,Virus ,Retrovirus ,Antigen ,medicine ,Immunology and Allergy ,Animals ,Humans ,Antigens, Viral ,biology ,General Medicine ,medicine.disease ,biology.organism_classification ,Virology ,Leukemia ,Macaca fascicularis ,Tumor Virus Infections ,medicine.anatomical_structure ,Humoral immunity ,Female ,Viral disease ,Retroviridae Infections - Abstract
The experimental infection of six cynomolgus monkeys with adult T-cell leukemia virus (ATLV) was attempted. Three animals were inoculated with living MT-2 cells and three with cell-free ATLV. All animals developed an antibody response to virus-specific glycopolypeptides and viral core polypeptides. ATLV-specific antigens appeared in peripheral lymphocytes from all six animals. Virus expression persisted in all animals. Up to 40 weeks after inoculation no animal developed any symptom of leukemia.
- Published
- 1984
34. Translation of HTLV (human T-cell leukemia virus) RNA in a nuclease-treated rabbit reticulocyte system
- Author
-
Naoki Yamamoto, Nobuyuki Kobayashi, Josef Schneider, Gerhard Hunsmann, Yoshio Koyanagi, and Masakazu Hatanaka
- Subjects
Adult ,Reticulocytes ,Antibodies, Neoplasm ,viruses ,T-Lymphocytes ,Biology ,Deltaretrovirus ,General Biochemistry, Genetics and Molecular Biology ,Virus ,Cell Line ,Viral Proteins ,Retrovirus ,Reticulocyte ,Antigens, Neoplasm ,Gene expression ,Protein biosynthesis ,medicine ,Animals ,Humans ,Molecular Biology ,Gel electrophoresis ,Leukemia ,Membrane Glycoproteins ,General Immunology and Microbiology ,Cell-Free System ,General Neuroscience ,RNA ,biology.organism_classification ,Virology ,Molecular biology ,medicine.anatomical_structure ,Retroviridae ,Protein Biosynthesis ,Agarose gel electrophoresis ,RNA, Viral ,Rabbits ,Research Article - Abstract
A human type-C retrovirus, designated HTLV (human T-cell leukemia virus), was isolated from the HTLV producer cell line MT-2. Agarose gel electrophoresis analysis 32P-labeled HTLVMT-2 virion RNA revealed that HTLVMT-2 virion RNA consists mainly of 24S and small amounts of 35S and 32S RNAs. The 24S HTLVMT-2 virion RNA and unfractionated HTLVMT-2 virion RNA were translated in a rabbit reticulocyte lysate system in vitro. The predominant polypeptide synthesized from 24S RNA had an apparent mol. wt. of 28 000 (28 K); unfractionated HTLVMT-2 virion RNA directed the synthesis of 53 000 (53 K), 33 000 (33 K) and 28 000 (28 K) polypeptides as main components. Most of the polypeptides synthesised in vitro by translation of HTLVMT-2 virion RNAs possess the same sizes as the proteins formerly designated as ATLA (ATL-associated antigen) in SDS-polyacrylamide gel electrophoresis and immunologically precipitated with sera of ATL patients. Therefore, the antigens termed ATLA, found by the serological study of ATL, are HTLVMT-2 encoded polypeptides.
- Published
- 1984
35. Antibodies to human T-cell leukemia virus types I and III in blood donors from Calabar, Nigeria
- Author
-
Josef Schneider, Robert A. Okpara, Gerhard Hunsmann, E.E. Williams, and Irene Wendler
- Subjects
Adult ,Male ,animal diseases ,viruses ,Captivity ,Nigeria ,Blood Donors ,Antibodies, Viral ,Cercopithecus aethiops ,Deltaretrovirus ,Virus ,West germany ,hemic and lymphatic diseases ,Internal Medicine ,Medicine ,Humans ,Leukemia ,biology ,business.industry ,virus diseases ,General Medicine ,medicine.disease ,Virology ,Human T cell leukemia virus ,Immunology ,biology.protein ,African Green Monkey ,Antibody ,business - Abstract
Excerpt To the editor: Antibodies to the human T-cell leukemia virus (HTLV) have been detected in sera of African green monkeys (Cercopithecus aethiops) which are in captivity in West Germany but w...
- Published
- 1986
36. A new ELISA test for HIV antibodies using a bacterially produced viral env gene product
- Author
-
J. Mous, H Gallati, Gerhard Hunsmann, D Stüber, I Wendler, F Guillot, H J Schoenfeld, and Josef Schneider
- Subjects
Microbiology (medical) ,Immunoprecipitation ,viruses ,Recombinant Fusion Proteins ,Immunology ,Enzyme-Linked Immunosorbent Assay ,HIV Antibodies ,Gp41 ,medicine.disease_cause ,Antibodies, Viral ,Gene product ,Antigen ,Viral Envelope Proteins ,Predictive Value of Tests ,medicine ,Immunology and Allergy ,Humans ,Escherichia coli ,Antigens, Viral ,chemistry.chemical_classification ,Acquired Immunodeficiency Syndrome ,biology ,virus diseases ,HIV ,General Medicine ,Virology ,Molecular biology ,Fusion protein ,Amino acid ,chemistry ,biology.protein ,Immunologic Techniques ,Antibody - Abstract
Screening tests for antibodies to the human immunodeficiency virus (HIV), based on the indirect ELISA principle using viral preparations as antigen, yield a substantial number of false-positive and false-negative results. These failures are due to the lack of certain viral polypeptides or contaminating cellular polypeptides in viral preparations. Therefore, the accuracy of the screening tests should be improved by using highly purified, synthetic viral antigens. With establishment of such an ELISA antigen in mind, we examined a bacterially synthesized polypeptide [ENV(80)] that corresponds to 80 conserved amino acids of the HIV gp41 transmembrane glycoprotein. ENV(80) was expressed as a DHFR fusion protein in Escherichia coli. Results obtained by HIV ELISA and immunoprecipitation with 497 serum samples from various groups at risk of AIDS were compared with those obtained with the ENV(80) ELISA. The ENV(80) ELISA was found to be superior to the H9/HTLV-III ELISA with respect to sensitivity and specificity and is almost equivalent in accuracy to immunoprecipitation.
- Published
- 1987
37. Immunoprevention of Friend virus-induced erythroleukemia by vaccination with viral envelope glycoprotein complexes
- Author
-
Josef Schneider, Annegret Schulz, and Gerhard Hunsmann
- Subjects
animal structures ,Viremia ,Antibodies, Viral ,Virus ,Mice ,Viral Proteins ,Viral envelope ,Antigen ,Viral Envelope Proteins ,Virology ,medicine ,Animals ,Glycoproteins ,Immunity, Cellular ,Leukemia, Experimental ,biology ,Friend virus ,Vaccination ,Radioimmunoassay ,Viral Vaccines ,medicine.disease ,biology.organism_classification ,Friend murine leukemia virus ,Leukemia ,Immunology ,biology.protein ,Leukemia, Erythroblastic, Acute ,Antibody - Abstract
Adult STU mice were repeatedly immunized with aggregates of Friend virus envelope glycoprotein gp85. The antigen was given emulsified in incomplete Freund's adjuvant. After the last boost, animals were challenged with a dose of FLV inducing fatal erythroleukemia in 90–100% of the controls. Twenty of twenty mice immunized with 100 μg of gp85 were completely protected, while three of seven animals vaccinated with 10 μg of gp85 died of leukemia. Smaller doses were ineffective, as was the adjuvant alone. Only animals that had developed a strong antibody response survived the FLV challenge. Serum antibodies of these animals reacted with type-, species-, and interspecies-specific determinants of gp85. Only a marginal cellular reactivity to gp85 was detected. The development of viremia was monitored by the use of radioimmunoassay for the internal viral core polypeptide p30. Animals completely protected were not viremic later than 7 days post-FLV infection. Those mice of the immunized groups which died in spite of an early antibody response showed a sharp decrease in serum antibodies to gp85 and a simultaneous appearance of virus in blood about 2–4 weeks before death from leukemia. Thus, the suppression of virus spread by antibody seems to be important for immunoprevention of this virus-induced leukemia.
- Published
- 1981
38. Lymphadenopathy and antibodies to HTLV-III in homosexual men. Clinical, laboratory and epidemiological features
- Author
-
R. Thommsen, Josef Schneider, H Bayer, I. Guggenmoos-Holzmann, J. R. Kalden, Gerhard Hunsmann, K. Ritter, U. Bienzle, K. Zwingenberger, C. H. Coester, and A. Uy
- Subjects
Adult ,Male ,medicine.medical_specialty ,Virus transmission ,viruses ,Sexual Behavior ,Context (language use) ,030204 cardiovascular system & hematology ,Antibodies, Viral ,Deltaretrovirus ,Serology ,03 medical and health sciences ,0302 clinical medicine ,Drug Discovery ,Epidemiology ,medicine ,Humans ,030212 general & internal medicine ,Life Style ,Lymphatic Diseases ,Genetics (clinical) ,Aged ,Acquired Immunodeficiency Syndrome ,biology ,Epidemiological Factors ,business.industry ,General Medicine ,Homosexuality ,Hepatitis B ,Middle Aged ,medicine.disease ,3. Good health ,Immunology ,biology.protein ,Molecular Medicine ,Antibody ,business ,Htlv iii ,Retroviridae Infections - Abstract
The study provides information on the epidemiology of HTLV-III infection and the lymphadenopathy syndrome (LAP) in 374 German homosexual men. Sexual contacts in the USA and rectal enemas before receptive anal intercourse are the main risk factors associated with virus transmission. HTLV-III seropositivity is significantly correlated with LAP. Prominent clinical signs are infreqquent. Immunological and haematological abnormalities are prevalent, and the retrovirus infection is frequently associated with serological markers of other viruses (hepatitis B, herpes group viruses). Lymphadenopathy as a manifestation of HTLV-III infection is discussed within the context of AIDS-related disorders.
- Published
- 1985
39. Human adult T-cell leukaemia virus is distinct from a similar isolate of Japanese monkeys
- Author
-
Friedrich W. Hirsch, Naoki Yamamoto, Gerhard Hunsmann, Yorio Hinuma, Toru Chosa, Morihisa Okada, and Josef Schneider
- Subjects
Antigenicity ,Japanese monkeys ,biology ,viruses ,Macaque ,Virology ,Deltaretrovirus ,Virus ,Molecular Weight ,Japanese macaque ,Viral Proteins ,hemic and lymphatic diseases ,biology.animal ,Animals ,Humans ,Macaca ,Adult T-cell leukaemia ,Human Virus ,Isoelectric Point ,Antigens, Viral ,Intracellular - Abstract
Summary We have compared the structural polypeptides of an adult T-cell leukaemia (ATL) virus (ATLV) isolate from a Japanese patient with ATL with those of a similar virus derived from a Japanese macaque monkey. Both are distinct but related entities. Their core polypeptides p19 could not be distinguished, but p24, another core polypeptide, and their envelope glycopolypeptides differ. The human virus directs the synthesis of a single intracellular glycopolypeptide, gp68, while the macaque virus specifies two such glycopolypeptides, gp57 and gp50. Furthermore, the glycopolypeptides of both viruses are serologically distinct. Thus, these viruses represent subtypes of the ATLV family and the macaque virus is apparently not involved in human ATL.
- Published
- 1984
40. Search for mammalian type-C retrovirus polypeptides on infected animal cells and human tumor cell lines using interspecies-reacting antibodies
- Author
-
Josef Schneider, Gerhard Hunsmann, and Friedrich Deinhardt
- Subjects
viruses ,Cross Reactions ,Feline leukemia virus ,Virus ,Pathology and Forensic Medicine ,Cell Line ,Viral Proteins ,Retrovirus ,Antigen ,Antigens, Neoplasm ,Neoplasms ,Animals ,Humans ,Molecular Biology ,Antigens, Viral ,Gammaretrovirus ,Glycoproteins ,Antiserum ,biology ,Cell Membrane ,Cell Biology ,General Medicine ,Neoplasms, Experimental ,biology.organism_classification ,Cytotoxicity Tests, Immunologic ,Virology ,Molecular biology ,Precipitin Tests ,Retroviridae ,Cell culture ,biology.protein ,Antibody - Abstract
Cells producing endogenous and exogenous type-C retroviruses of murine, feline and primate origin were evaluated for expression of those virus-specific cell-surface antigens which cross-react with antibodies to interspecies determinants of mammalian type-C viral polypeptides. Surface polypeptides of cells replicating endogenous and exogenous type-C retroviruses were iodinated by the lactoperoxidase method. Labelled antigens were immunoprecipitated and analyzed in polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. This method detected gp71 and less frequently p15E/p12E on the surface of virus-producing cells; in addition, p27 was found on F422 cells replicating feline leukemia virus. Antibodies to the membrane protein p15E displayed the broadest cross-reactivity but only antibodies to gp71 mediated complement-dependent interspecies cell lysis. The pattern of cross-reactivity reflected known genetic relationships among these mammalian viruses. Although antiserum to the simian sarcoma virus complex (SSV) was strongly cytotoxic to some human tumor cell lines, this reactivity could not be attributed to antibodies cross-reacting with SSV gp71.
- Published
- 1980
41. Expression of HTLV-specific polypeptides in various human T-cell lines
- Author
-
Josef Schneider, Toru Chosa, Nobuyuki Kobayashi, Masakazu Hatanaka, Gerhard Hunsmann, Naoki Yamamoto, Yoshio Koyanagi, and Yorio Hinuma
- Subjects
Microbiology (medical) ,T cell ,T-Lymphocytes ,Immunology ,Biology ,Deltaretrovirus ,Virus ,Cell Line ,Viral Proteins ,Viral envelope ,Antigen ,Viral Envelope Proteins ,medicine ,Immunology and Allergy ,Humans ,Protein Precursors ,Antigens, Viral ,chemistry.chemical_classification ,Viral Core Proteins ,General Medicine ,medicine.disease ,Virology ,Leukemia, Lymphoid ,Leukemia ,medicine.anatomical_structure ,chemistry ,Cell culture ,Glycoprotein ,Peptides ,Intracellular ,Retroviridae Infections - Abstract
The adult T-cell leukemia (ATL)-associated antigen complex (ATLA) was first discovered with indirect immunofluorescence by Hinuma et al. (1981). Biochemical analysis with MT-2 cells revealed that ATLA consisted mainly of human T-cell leukemia virus (HTLV) structural polypeptides and their precursors (Yamamoto and Hinuma 1982a; Schneider et al. 1984). In this study, we have investigated the molecular nature of the ATLA antigen complex in various HTLV-positive human cell lines established by different methods including independently established HTLV-infected HUT 102 cells. We found that HTLVs infecting these cell lines have similar core polypeptides, p24 and p19, as well as an envelope glycopolypeptide, gp46, in all these cells. The intracellular gp61 and p53 appear to be precursors of the viral envelope and core polypeptides, respectively. Interestingly, MT-2 and MT-2 related T-cell lines contain two different species of envelope proteins, gp68 and gp61, whereas cell lines not related to MT-2 express only gp61.
- Published
- 1984
42. Antibodies to HTLV-I in Nigerian blood-donors, their relatives and patients with leukaemias, lymphomas and other diseases
- Author
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H Bayer, S. R. Bhusnurmath, R. Maharajah, Josef Schneider, A. G. Kulkarni, R. A. Okpara, E.E. Williams, I. Akinsete, AlanF. Fleming, Gerhard Hunsmann, and M. Abraham
- Subjects
Male ,Cancer Research ,medicine.medical_specialty ,Lymphoma ,Nigeria ,Blood Donors ,Antibodies, Viral ,Deltaretrovirus ,Virus ,Serology ,Acquired immunodeficiency syndrome (AIDS) ,immune system diseases ,hemic and lymphatic diseases ,parasitic diseases ,Epidemiology ,medicine ,Humans ,Leukemia ,biology ,business.industry ,Deltaretrovirus Antibodies ,medicine.disease ,Oncology ,Immunology ,biology.protein ,Female ,Viral disease ,Antibody ,business - Abstract
Antibodies to HTLV-I have been detected in sera from 15 (2.0%) of 736 adult blood-donors in Nigeria, in 4 (20.0%) of 20 patients with chronic lymphatic leukaemia, 3 (10.0%) of 30 with non-Hodgkin's lymphoma, one of 12 with Burkitt's lymphoma and one of 7 with acute lymphoblastic leukaemia. The frequency of positivily was higher (3.6%) in the blood-donors from the guinea and wooded savanna of northern Nigeria than in those from the rain-forest and mangrove swamps of southern Nigeria (1.8% in Lagos and 0.7% in Calabar). Two of the 3 seropositive patients with lymphoma had clinical presentation and courses similar to those of Japanese and Caribbean patients with adult T-cell leukaemia/lymphoma.
- Published
- 1986
43. Precursor polypeptides of adult T-cell leukaemia virus: detection with antisera against isolated polypeptides gp68, p24 and p19
- Author
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Gerhard Hunsmann, Josef Schneider, Yorio Hinuma, and Naoki Yamamoto
- Subjects
Antiserum ,biology ,viruses ,Nucleic acid sequence ,Provirus ,Group-specific antigen ,Antibodies, Viral ,Virology ,Molecular biology ,Deltaretrovirus ,Virus ,Gene product ,Molecular Weight ,chemistry.chemical_compound ,Viral Proteins ,Biosynthesis ,chemistry ,Antibody Specificity ,biology.protein ,Humans ,Electrophoresis, Polyacrylamide Gel ,Antibody ,Protein Precursors ,Glycoproteins - Abstract
Summary Sera of individuals infected with adult T-cell leukaemia virus (ATLV) react predominantly with the polypeptides gp68, p24 and p19. These polypeptides were isolated from ATLV-infected MT-2 cells and virus. The radioiodinated polypeptides were used to quantify respective antibodies in individual ATLV carrier sera. Heteroantisera prepared in rabbits against isolated polypeptides facilitated studies on the biosynthesis of the core and envelope polypeptides of ATLV. Pulse-chase experiments revealed a polypeptide of mol. wt. 48000 (48K) as the precursor to the core polypeptides p24 and p19. A 28K polypeptide related to p19 appeared to be an early side-product of the gag gene or a translate of a defective viral message. Antiserum to the putative env gene product gp68 recognized gp68, gp66 and small amounts of gp62. In tunicamycin-treated cells gp68, gp66 and gp62 were no longer synthesized, but a 54K polypeptide reacted with antiserum to gp68. Polypeptide p54 is structurally related to gp68 and therefore apparently represents the unglycosylated form of gp68. Moreover, the apparent mol. wt. of p54 and p48 agree with those predicted for respective env and gag precursors from the nucleotide sequence of an ATLV provirus.
- Published
- 1984
44. Lymphoma-specific T- and B-cell responses suggest the involvement of HTLV-I in virus-non-productive lymphomas of a married couple
- Author
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Schlote W, Mann Dl, E M Schneider, Peter Wernet, Josef Schneider, Saal Jg, and Graham Pawelec
- Subjects
Male ,Cancer Research ,T-Lymphocytes ,Biology ,Antibodies, Viral ,Lymphocyte Activation ,Virus ,Immune system ,HLA Antigens ,medicine ,Cytotoxic T cell ,Humans ,B cell ,Aged ,B-Lymphocytes ,Lymphokine-activated killer cell ,Deltaretrovirus Antibodies ,Lymphoma, Non-Hodgkin ,T lymphocyte ,HLA-DR Antigens ,medicine.disease ,HTLV-I Infections ,Lymphoma ,medicine.anatomical_structure ,Phenotype ,Oncology ,Immunology ,Female ,Clone (B-cell biology) - Abstract
The occurrence of aberrant cellular progeny is not an infrequent event in vivo. Viral infections (Hanto et al., 1981; Gallo, 1984) and/or failures in genetic programming during cell proliferarion may facilitate the manifestation of malignancy in states of incomplete immune surveillance. This is suggested by a high incidence of lymphoid malignancies in immunode-pressed individuals, as for example after organ transplantation (Penn, 1981). Efficient cellular defense mechanisms which control malignant growth have been postulated. So far, MHC-restricted, tumor-specific cytotoxic T cells have not been identified in vivo. Moreover, effector cells with natural killer activity have not consistently been shown to function as general or specific anti-tumor agents in vivo. Nevertheless, LAK cells are presently under investigation in adoptive immuno-therapy for tumors. The concept of LAK-cell treatment is based on the observation that high concentrations of interleu-kin 2 (IL-2) induce activation and proliferation of autologous, tumor-specific cytotoxic cells (Hersey et al., 1981; Lotze et al., 1981). The most promising results have been obtained in mouse tumor models, whereas treatment of human diseases has rarely been successful (Rosenberg et al., 1986). In the present report, high percentages of CD25-positive T lymphocytes were detected in PB and LN cell suspensions of a patient with a B-cell NHL. These T cells were successfully enriched in cultures containing low doses of IL-2. The CTX mediated by these T-cell lines and a single T-cell clone derived from them was specific for autologous lymphoma cells as well as for the lymphoma cells from a different individual. Interestingly, an HTLV-I-infected tumor-cell line was recognized also. The specific cellular as well as the humoral immune response. however, was drastically reduced following chemo- and radiotherapy. In contrast, non-specific CTX mediated by aclivated NK-like cells persisted.
- Published
- 1988
45. Seroepidemiology of HTLV-III (LAV) in the Federal Republic of Germany
- Author
-
Gerhard Hunsmann, E. Lechler, Peter Hellstern, Peter Kern, R. Hehlmann, Volker Erfle, E. M. Schneider, G. Krüger, W. Kreuz, H. D. Brede, H Bayer, U. Egli, L. Bergmann, Erhard Seifried, Josef Schneider, E. Holzer, R. Kurth, W. Schneider, H. R. Brodt, Herbert Schmitz, I. Helm, U. Bienzle, K. Schimpf, Frank-D. Goebel, I. Scharrer, Peter Wernet, H. Rasokat, H. Berthold, Albrecht Werner, and W. Mellert
- Subjects
Male ,Risk ,Drug abuser ,Enzyme-Linked Immunosorbent Assay ,Antibodies, Viral ,Deltaretrovirus ,Virus ,Risk groups ,Acquired immunodeficiency syndrome (AIDS) ,Drug Discovery ,Humans ,Medicine ,Genetics (clinical) ,Acquired Immunodeficiency Syndrome ,biology ,business.industry ,Germany, West ,Federal republic of germany ,Homosexuality ,General Medicine ,biology.organism_classification ,medicine.disease ,Virology ,Immunology ,Acquired Immunodeficiency Syndrome (aids) ,Lymphadenopathy-associated Virus (lav)/human T-lymphotropic Virus Type Iii (htlv-iii) ,Seroepidemiology ,biology.protein ,Molecular Medicine ,Female ,Antibody ,business ,Htlv iii ,Retroviridae Infections - Abstract
In 1984 10,281 sera were collected in the FRG and examined for antibodies to HTLV-III (LAV) with an enzyme-linked immunosorbent assay and confirmative tests. Of the German AIDS patients 81% have antibodies. Individuals belonging to AIDS risk groups, homosexuals, haemophiliacs and i.v. drug abusers, have antibody frequencies between 25%–72%. The detection of HTLV-III antibodies in blood donours indicates that the virus is being transmitted by blood transfusions.
- Published
- 1985
46. Characterization of African green monkey B-cell lines releasing an adult T-cell leukemia-virus-related agent
- Author
-
Naoki Yamamoto, Josef Schneider, Gerhard Hunsmann, Masakazu Hatanaka, Nobuyuki Kobayashi, Kaoru Takeuchi, Toru Chosa, Yorio Hinuma, and Yoshio Koyanagi
- Subjects
Cancer Research ,Herpesvirus 4, Human ,T-cell leukemia ,Cercopithecus ,Immunofluorescence ,Deltaretrovirus ,Virus ,Cell Line ,Antigen ,Chlorocebus aethiops ,medicine ,Animals ,Humans ,Antigens, Viral ,B cell ,B-Lymphocytes ,medicine.diagnostic_test ,biology ,medicine.disease ,Cell Transformation, Viral ,Virology ,Leukemia ,medicine.anatomical_structure ,Retroviridae ,Oncology ,Cell culture ,DNA, Viral ,biology.protein ,Antibody - Abstract
Eight lymphoblastoid cell lines were established from the peripheral blood of individual African green monkeys (AGM). The AGM-2206 line grew out spontaneously. The others - AGM-6, 7, 8, 10, 12, 13, and 16 - were obtained after infection of peripheral AGM lymphocytes with cell-free culture supernatant of AGM-2206. All lines contained, and were probably transformed by, AGM-EBV. Moreover, they expressed immunoglobulins but lacked the Leu l T-cell marker. Thus they were B cells. Since a high percentage of AGMs are naturally infected with a virus similar to adult T-cell leukemia virus (ATLV), we examined these cell lines for ATLV. With immunofluorescence tests we detected ATLV-related antigens (ATLA) in three of the eight cell lines. EBV membrane antigen was present in three out of four. The highest percentage (40%) of ATLA-positive cells was found in the AGM-13 line. After metabolic labelling of these cells, ATLV-specific polypeptides p24, p19, p15, and p10 were detected. Hybridization experiments showed that both AGM-2206 and AGM-13 cell lines contained ATLV-proviral DNA. Electron micrographs of AGM-13 revealed a few type-C particles morphologically similar to the MT-2 virus. By cocultivation this AGM virus was able to infect and immortalize human peripheral blood lymphocytes. One such human cell line, NA-13, expressed polypeptides closely related to ATLV core antigens but a 68,000 mol.wt. glycopolypeptide was serologically distinct from MT-2 ATLV gp68.
- Published
- 1984
47. Properties of retroviral protease responsible for gag precursor cleavage
- Author
-
Rudolf Hafenrichter, Heinz-Jürgen Thiel, Josef Schneider, and Frank Weiland
- Subjects
Macromolecular Substances ,viruses ,Proteolysis ,medicine.medical_treatment ,Detergents ,Retroviridae Proteins ,Gene Products, gag ,Cleavage (embryo) ,Virus ,Viral Proteins ,Virology ,Murine leukemia virus ,medicine ,chemistry.chemical_classification ,Protease ,medicine.diagnostic_test ,biology ,Temperature ,biology.organism_classification ,Molecular biology ,In vitro ,Cell Compartmentation ,Friend murine leukemia virus ,NS2-3 protease ,Molecular Weight ,Enzyme ,Biochemistry ,chemistry ,Protein Processing, Post-Translational ,Peptide Hydrolases - Abstract
Friend Leukemia Virus (FLV) particles contain a protease which cleaves not only its own gag precursor but also the gag polyprotein of simian sarcoma virus (SSV). To determine the localization of the enzyme within the virion, purified virus was fractionated. According to our studies most of the proteolytic activity is located within the retroviral core. Since ionic detergents inhibit the protease, heat treatment was found to be the most effective way to release the molecule from the virus particle. The in vitro studies indicate a high resistance of the enzyme to different kinds of heat treatment.
- Published
- 1989
48. Experimental infection of rhesus monkeys with SIV isolated from African green monkeys
- Author
-
Ottmar Herchenröder, Christiane Stahl-Hennig, Wolfgang Lüke, Josef Schneider, Gabriele Schulze, Heinz Hartmann, Heidemarie Schmidt, Klara Tenner-Racz, Paul Racz, Masanori Hayami, John C. Kelliher, and Gerhard Hunsmann
- Subjects
Acquired Immunodeficiency Syndrome ,Cercopithecidae ,Simian immunodeficiency virus ,Biology ,Pathogenicity ,medicine.disease_cause ,medicine.disease ,Antibodies, Viral ,Virology ,Macaca mulatta ,Disease Models, Animal ,Infectious Diseases ,Animal model ,Acquired immunodeficiency syndrome (AIDS) ,medicine ,Juvenile ,Animals ,Macaca ,Simian Immunodeficiency Virus ,African Green Monkey ,Lymph Nodes ,Viremia ,Retroviridae Infections - Abstract
To establish an animal model for AIDS, 6 juvenile rhesus monkeys were infected intravenously with cell-free SIVagmTYO-1, 5, or 7. One animal was infected with SIV of known pathogenicity isolated from a sooty mangabey (SIVsmm). The 2 animals infected with TYO-1, 1 of 2 infected with TYO-7 and the 1 infected with SIVsmm seroconverted within 4-8 weeks after infection and infectious virus could be recovered 8-10 or 44 weeks after infection. Three of the 4 seroconverted monkeys developed a persistent lymphadenopathy 12 weeks after infection. Rhesus monkeys infected with SIVagm could serve as a model to study HIV infection and for vaccine trials.
- Published
- 1989
49. Sera from adult T-cell leukemia patients react with envelope and core polypeptides of adult T-cell leukemia virus
- Author
-
Naoki Yamamoto, Josef Schneider, Yorio Hinuma, and Gerhard Hunsmann
- Subjects
T-Lymphocytes ,T-cell leukemia ,Fluorescent Antibody Technique ,Immunofluorescence ,Antibodies, Viral ,Deltaretrovirus ,Virus ,Viral Proteins ,Antigen ,Viral envelope ,Viral Envelope Proteins ,Virology ,medicine ,Humans ,Immunosorbent Techniques ,Leukemia ,medicine.diagnostic_test ,biology ,Viral Core Proteins ,medicine.disease ,Molecular biology ,Kinetics ,Cell culture ,Concanavalin A ,biology.protein - Abstract
Sera from five Japanese patients with adult T-cell leukemia (ATL) showed in the immunofluorescence test for ATL-associated antigen (ATLA) titers ranging from 320 to 1280. Control sera from three healthy adults were negative. These eight sera were used to immunoprecipitate radiolabeled polypeptides from three cell lines infected with adult T-cell leukemia virus (ATLV) and two noninfected human cell lines. Cells were labeled either metabolically with [35S]cysteine, [35S]methionine, or [3H]glucosamine, or chemically with 125-iodine. Immunoprecipitates from cells, virus, and concanavalin A-enriched supernatants were analyzed by polyacrylamide gel electrophoresis. Cells producing ATLV contain gp68, the putative precursor to ATLV envelope polypeptides, gp46, and possibly p15, in addition to several nonglycosylated polypeptides between 40 to 70 kDa. Gp46 is shed into the culture medium and appears to be loosely attached to the viral and cellular surface. After purification on density gradients viral particles contain immunoreactive p24, p19, p15, and small amounts of gp46. Kinetics of synthesis, distribution, size, biochemical characteristics, and immunoreactivity of these polypeptides strongly suggest that most of them are structural components of ATLV or their precursors. Apparently, the intracellular ATLA complex predominantly represents precursors of viral structural polypeptides and gp46 is the viral envelope glycopolypeptide.
- Published
- 1984
50. Adult T-cell leukemia (ATL) virus-specific antibodies in ATL patients and healthy virus carriers
- Author
-
Gerhard Hunsmann, Naoki Yamamoto, Josef Schneider, Yoshio Koyanagi, and Yorio Hinuma
- Subjects
Cancer Research ,Antibodies, Neoplasm ,T-Lymphocytes ,T-cell leukemia ,Biology ,Immunofluorescence ,Antibodies, Viral ,Virus ,Gene product ,Viral Proteins ,Antigen ,medicine ,Humans ,Glycoproteins ,Leukemia ,medicine.diagnostic_test ,medicine.disease ,Virology ,Titer ,Retroviridae ,Oncology ,Immunology ,Carrier State ,biology.protein ,Antibody - Abstract
The adult T-cell leukemia (ATL)-associated antigen complex (ATLA) is recognized by serum antibodies of carriers of ATL virus (ATLV). ATLA consists mainly of ATLV polypeptides and their precursors. The sera from 22 ATL patients, 21 healthy carriers and 9 healthy individuals were examined quantitatively by immunofluorescence assay (IF) for ATLA and by a newly developed radioimmunoprecipitation test with purified 125I-gp68, the putative env gene product of ATLV. More qualitative results were obtained by analysis on polyacrylamide gel (PAGE) of immunoprecipitates from lysates of 35-S-cysteine-la-belled cells producing ATLV, pelleted ATLV and cell-free culture supernatant. The two quantitative assays gave negative results with sera from all normal subjects and a few patients, but detected ATLA antibodies in all the healthy ATLV carriers. An important finding was that sera of patients that gave negative results in one assay gave positive, results in the other, and vice versa. In contrast, all sera from ATL patients and healthy carriers, but not normal donors, precipitated ATLV-specific glycopolypeptides, gp68 and gp46 from 35S-labelled materials. But core polypeptides p28, p24, p19 and p15 were precipitated only by sera with IF titers of over 80. Thus, anti-ATLA antibodies in seropositive sera are predominantly directed against glycopolypeptides of ATLV, and the antibody reactivity to ATLA antigens does not differentiate between ATL patients at various stages of the disease and healthy ATLV carriers.
- Published
- 1983
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