Neopterin and Nitrite in Supernatants from Interferon-γ-treated Monocytoid Cell Lines: A Tool to Identify Bacterial Pyrogens
The rabbit pyrogen assay identifies pyrogenic contaminations in drugs i.l1tend~d for parenteral use. In order to replace this test because of ethical and economical reasons, various in vitro methods have been developed in recent years. Here, we summarize our results on optimizing a test based on usi...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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De Gruyter
1999-08-01
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| Series: | Pteridines |
| Subjects: | |
| Online Access: | https://doi.org/10.1515/pteridines.1999.10.3.112 |
| _version_ | 1818731984968482816 |
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| author | Peterbauer Anja Werner Ernst R. Werner-Felmayer Gabriele |
| author_facet | Peterbauer Anja Werner Ernst R. Werner-Felmayer Gabriele |
| author_sort | Peterbauer Anja |
| collection | DOAJ |
| description | The rabbit pyrogen assay identifies pyrogenic contaminations in drugs i.l1tend~d for parenteral use. In order to replace this test because of ethical and economical reasons, various in vitro methods have been developed in recent years. Here, we summarize our results on optimizing a test based on using interferony- treated monocytoid cell lines from man (THP-l) and mouse (RAW264.7). The read-out of the test is neopterin or nitrite, respectively, which is released into the supernatant in response to bacterial compounds. These are costimuli of the enzymes involved in neopterin and nitrite formation, i.e. GTP cyclohydrolase I and inducible nitric oxide synthase. The test reproducibly detects cell wall components from Gram-negative as well as from Gram-positive bacteria and mycobacteria. Furthermore, DNA and cell-free supernatants containing a number of bioactive but not further characterized proteins, both from Staphylococcus aureus, can be detected. Thus, this test is superior over the only in vitro alternative accepted in certain cases, namely the limulus amebocyte lysate assay. Results obtained by measuring neopterin or nitrite correlate well with formation of the endogenous pyrogen tumor necrosis factor-a, a read-out used by some other cell-based in vitro alternatives to the rabbit pyrogen test. We therefore think that the assay presented here has the potential to reduce or even replace the animal test. |
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| id | doaj.art-90dfec6e419d4c3e8631ad60429764fa |
| institution | Directory Open Access Journal |
| issn | 0933-4807 2195-4720 |
| language | English |
| last_indexed | 2024-12-17T23:26:22Z |
| publishDate | 1999-08-01 |
| publisher | De Gruyter |
| record_format | Article |
| series | Pteridines |
| spelling | doaj.art-90dfec6e419d4c3e8631ad60429764fa2022-12-21T21:28:45ZengDe GruyterPteridines0933-48072195-47201999-08-0110311211810.1515/pteridines.1999.10.3.112Neopterin and Nitrite in Supernatants from Interferon-γ-treated Monocytoid Cell Lines: A Tool to Identify Bacterial PyrogensPeterbauer Anja0Werner Ernst R.1Werner-Felmayer Gabriele2Institute of Medical Chemistry and Biochemistry, University of Innsbruck, Fritz-Pregl-Strasse 3, A-6020 Innsbruck, AustriaInstitute of Medical Chemistry and Biochemistry, University of Innsbruck, Fritz-Pregl-Strasse 3, A-6020 Innsbruck, AustriaInstitute of Medical Chemistry and Biochemistry, University of Innsbruck, Fritz-Pregl-Strasse 3, A-6020 Innsbruck, AustriaThe rabbit pyrogen assay identifies pyrogenic contaminations in drugs i.l1tend~d for parenteral use. In order to replace this test because of ethical and economical reasons, various in vitro methods have been developed in recent years. Here, we summarize our results on optimizing a test based on using interferony- treated monocytoid cell lines from man (THP-l) and mouse (RAW264.7). The read-out of the test is neopterin or nitrite, respectively, which is released into the supernatant in response to bacterial compounds. These are costimuli of the enzymes involved in neopterin and nitrite formation, i.e. GTP cyclohydrolase I and inducible nitric oxide synthase. The test reproducibly detects cell wall components from Gram-negative as well as from Gram-positive bacteria and mycobacteria. Furthermore, DNA and cell-free supernatants containing a number of bioactive but not further characterized proteins, both from Staphylococcus aureus, can be detected. Thus, this test is superior over the only in vitro alternative accepted in certain cases, namely the limulus amebocyte lysate assay. Results obtained by measuring neopterin or nitrite correlate well with formation of the endogenous pyrogen tumor necrosis factor-a, a read-out used by some other cell-based in vitro alternatives to the rabbit pyrogen test. We therefore think that the assay presented here has the potential to reduce or even replace the animal test.https://doi.org/10.1515/pteridines.1999.10.3.112pyrogenalternative test methodmonocytoid cell linesneopterinnitritelimulus amebocyte lysate assay |
| spellingShingle | Peterbauer Anja Werner Ernst R. Werner-Felmayer Gabriele Neopterin and Nitrite in Supernatants from Interferon-γ-treated Monocytoid Cell Lines: A Tool to Identify Bacterial Pyrogens Pteridines pyrogen alternative test method monocytoid cell lines neopterin nitrite limulus amebocyte lysate assay |
| title | Neopterin and Nitrite in Supernatants from Interferon-γ-treated Monocytoid Cell Lines: A Tool to Identify Bacterial Pyrogens |
| title_full | Neopterin and Nitrite in Supernatants from Interferon-γ-treated Monocytoid Cell Lines: A Tool to Identify Bacterial Pyrogens |
| title_fullStr | Neopterin and Nitrite in Supernatants from Interferon-γ-treated Monocytoid Cell Lines: A Tool to Identify Bacterial Pyrogens |
| title_full_unstemmed | Neopterin and Nitrite in Supernatants from Interferon-γ-treated Monocytoid Cell Lines: A Tool to Identify Bacterial Pyrogens |
| title_short | Neopterin and Nitrite in Supernatants from Interferon-γ-treated Monocytoid Cell Lines: A Tool to Identify Bacterial Pyrogens |
| title_sort | neopterin and nitrite in supernatants from interferon γ treated monocytoid cell lines a tool to identify bacterial pyrogens |
| topic | pyrogen alternative test method monocytoid cell lines neopterin nitrite limulus amebocyte lysate assay |
| url | https://doi.org/10.1515/pteridines.1999.10.3.112 |
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