Serological characterization of anti-endotoxin serum directed against the conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid
The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The neoglycoconjugate was a good immunogen in rabbits yielding a high level of anti-lipopolysaccharide (LPS) antibodies of the IgG class. It was found th...
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| Published in | FEMS immunology and medical microbiology Vol. 37; no. 1; pp. 59 - 67 |
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| Main Authors | , , , , , , , , , |
| Format | Journal Article |
| Language | English |
| Published |
Oxford, UK
Elsevier B.V
10.06.2003
Blackwell Publishing Ltd Blackwell |
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| Abstract | The covalent conjugate of oligosaccharide core of
Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The neoglycoconjugate was a good immunogen in rabbits yielding a high level of anti-lipopolysaccharide (LPS) antibodies of the IgG class. It was found that antiserum was able to react with the smooth LPS molecules of identical (R4) or related (R1) core type. The reactions were shown in the enzyme-linked immunosorbent assay and the immunoblotting test. Flow cytometry showed that anti-core antibodies reacted with LPS present on intact, live, smooth bacteria labelling more than 90% of cells. The anti-OS R4-TT serum used for in vitro studies showed high endotoxin neutralization activity. The serum inhibited endotoxin-induced tumor necrosis factor α and nitric oxide synthesis by the J-774A.1 cell line and attenuated pulmonary retention of YAC-1 cells. |
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| AbstractList | The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The neoglycoconjugate was a good immunogen in rabbits yielding a high level of anti-lipopolysaccharide (LPS) antibodies of the IgG class. It was found that antiserum was able to react with the smooth LPS molecules of identical (R4) or related (R1) core type. The reactions were shown in the enzyme-linked immunosorbent assay and the immunoblotting test. Flow cytometry showed that anti-core antibodies reacted with LPS present on intact, live, smooth bacteria labelling more than 90% of cells. The anti-OS R4-TT serum used for in vitro studies showed high endotoxin neutralization activity. The serum inhibited endotoxin-induced tumor necrosis factor alpha and nitric oxide synthesis by the J-774A.1 cell line and attenuated pulmonary retention of YAC-1 cells. The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The neoglycoconjugate was a good immunogen in rabbits yielding a high level of anti‐lipopolysaccharide (LPS) antibodies of the IgG class. It was found that antiserum was able to react with the smooth LPS molecules of identical (R4) or related (R1) core type. The reactions were shown in the enzyme‐linked immunosorbent assay and the immunoblotting test. Flow cytometry showed that anti‐core antibodies reacted with LPS present on intact, live, smooth bacteria labelling more than 90% of cells. The anti‐OS R4‐TT serum used for in vitro studies showed high endotoxin neutralization activity. The serum inhibited endotoxin‐induced tumor necrosis factor α and nitric oxide synthesis by the J‐774A.1 cell line and attenuated pulmonary retention of YAC‐1 cells. Abstract The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The neoglycoconjugate was a good immunogen in rabbits yielding a high level of anti-lipopolysaccharide (LPS) antibodies of the IgG class. It was found that antiserum was able to react with the smooth LPS molecules of identical (R4) or related (R1) core type. The reactions were shown in the enzyme-linked immunosorbent assay and the immunoblotting test. Flow cytometry showed that anti-core antibodies reacted with LPS present on intact, live, smooth bacteria labelling more than 90% of cells. The anti-OS R4-TT serum used for in vitro studies showed high endotoxin neutralization activity. The serum inhibited endotoxin-induced tumor necrosis factor α and nitric oxide synthesis by the J-774A.1 cell line and attenuated pulmonary retention of YAC-1 cells. The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The neoglycoconjugate was a good immunogen in rabbits yielding a high level of anti-lipopolysaccharide (LPS) antibodies of the IgG class. It was found that antiserum was able to react with the smooth LPS molecules of identical (R4) or related (R1) core type. The reactions were shown in the enzyme-linked immunosorbent assay and the immunoblotting test. Flow cytometry showed that anti-core antibodies reacted with LPS present on intact, live, smooth bacteria labelling more than 90% of cells. The anti-OS R4-TT serum used for in vitro studies showed high endotoxin neutralization activity. The serum inhibited endotoxin-induced tumor necrosis factor α and nitric oxide synthesis by the J-774A.1 cell line and attenuated pulmonary retention of YAC-1 cells. |
| Author | Jachymek, Wojciech Lakomska, Joanna Fortuna, Wojciech Niedziela, Tomasz Dzieciatkowska, Monika Lukasiewicz, Jolanta Miedzybrodzki, Ryszard Szymaniec, Stanislaw Lugowski, Czeslaw Misiuk-Hojlo, Marta |
| Author_xml | – sequence: 1 givenname: Jolanta surname: Lukasiewicz fullname: Lukasiewicz, Jolanta email: czaja@iitd.pan.wroc.pl organization: Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland – sequence: 2 givenname: Wojciech surname: Jachymek fullname: Jachymek, Wojciech organization: Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland – sequence: 3 givenname: Tomasz surname: Niedziela fullname: Niedziela, Tomasz organization: Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland – sequence: 4 givenname: Monika surname: Dzieciatkowska fullname: Dzieciatkowska, Monika organization: Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland – sequence: 5 givenname: Joanna surname: Lakomska fullname: Lakomska, Joanna organization: Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland – sequence: 6 givenname: Ryszard surname: Miedzybrodzki fullname: Miedzybrodzki, Ryszard organization: Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland – sequence: 7 givenname: Wojciech surname: Fortuna fullname: Fortuna, Wojciech organization: Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland – sequence: 8 givenname: Stanislaw surname: Szymaniec fullname: Szymaniec, Stanislaw organization: Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland – sequence: 9 givenname: Marta surname: Misiuk-Hojlo fullname: Misiuk-Hojlo, Marta organization: Department of Ophthalmology, Medical University of Wroclaw, ul. Chalubinskiego 2a, 50-368 Wroclaw, Poland – sequence: 10 givenname: Czeslaw surname: Lugowski fullname: Lugowski, Czeslaw organization: Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wroclaw, Poland |
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| Keywords | R4 Antibody Endotoxin Escherichia coli Neoglycoconjugate Core region Enzyme Oligosaccharide Metalloendopeptidases Peptidases Toxin Lipopolysaccharide Bacteria Hydrolases Serum Tentoxilysin Enterobacteriaceae |
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| Snippet | The covalent conjugate of oligosaccharide core of
Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The... Abstract The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The... The covalent conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid was prepared using reaction of reductive amination. The... |
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| SubjectTerms | Animals Antibody Bacteriology Biological and medical sciences Carbohydrate Sequence Cell Line Core region Endotoxin Endotoxins - immunology Enzyme-Linked Immunosorbent Assay Escherichia coli Escherichia coli - chemistry Escherichia coli - immunology Flow Cytometry Fundamental and applied biological sciences. Psychology Glycoconjugates - immunology Immune Sera - immunology Immunoblotting Lipopolysaccharides - chemistry Lipopolysaccharides - immunology Lung - cytology Lung - immunology Macrophages - immunology Mice Microbiology Molecular Sequence Data Neoglycoconjugate Nitric Oxide - biosynthesis Nitric Oxide - immunology Tetanus Toxoid - chemistry Tetanus Toxoid - immunology Tumor Necrosis Factor-alpha - biosynthesis Tumor Necrosis Factor-alpha - immunology Vaccines, antisera, therapeutical immunoglobulins and monoclonal antibodies |
| Title | Serological characterization of anti-endotoxin serum directed against the conjugate of oligosaccharide core of Escherichia coli type R4 with tetanus toxoid |
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