The International Standard for Endotoxin: evaluation in an international collaborative study

Poole, Stephen; Mussett, Marjorie V.

doi:10.1016/0092-1157(89)90006-1

Cited by 32 publications

(18 citation statements)

References 4 publications

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“…In another study, Eperon and Jungi (10) obtained a detection limit of 15 pg/ml with calcitriol-primed MM6 subclones, a value comparable to that for MM6-CA8 in this study (9.6 pg/ml). For comparison, the sensitivity of the rabbit pyrogen test is approximately 1 ng/ml (27). Eperon et al (9) proved that MM6 cells are sensitive to lipopolysaccharide (endotoxin) of various origins.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the In Vitro Pyrogen Test System Based on Proinflammatory Cytokine Release from Human Monocytes: Comparison with a Human Whole Blood Culture Test System and with the Rabbit Pyrogen Test

Nakagawa

Maeda

Murai

2002

Clin Vaccine Immunol

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The reliability of an in vitro pyrogen test system based on proinflammatory cytokine release from human monocytic cells was assessed by comparison with a test system based on a human whole blood culture as well as with the conventional rabbit pyrogen test. The human cells used as the pyrogen indicator cells were newly selected by subcloning of a human monocytic cell line, Mono-Mac-6. The selected cells, named MM6-CA8, responded to various pyrogens, including endotoxin, peptidoglycan (PG), Staphylococcus aureus Cowan 1 (SAC), and poly(I · C), with a high sensitivity and produced proinflammatory cytokines, such as interleukin 1 (IL-1), IL-6, and tumor necrosis factor alpha. Among these cytokines, IL-6 was produced most sensitively in response to traces of the pyrogens and detected in the largest quantities in the culture medium. The cytokineproducing responses of MM6-CA8 cells correlated significantly with the responses of cultured human whole blood, which represents an ex vivo culture test system reproducing pyrogen-induced cytokine production in the human body. In terms of cytokine inducibility, the pyrogens were ranked in the order endotoxin > PG > poly (I · C) > SAC in both culture systems, a ranking which almost agreed with the ranking of their pyrogenicity as assessed by the rabbit pyrogen test. These results suggest that the in vitro responsiveness of MM6-CA8 cells to various pyrogens is highly relevant for human pyrogenic reactions. Therefore, the in vitro test system is useful and reliable for detecting the presence of materials that are pyrogenic for humans.

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Section: Discussionmentioning

confidence: 99%

Evaluation of the In Vitro Pyrogen Test System Based on Proinflammatory Cytokine Release from Human Monocytes: Comparison with a Human Whole Blood Culture Test System and with the Rabbit Pyrogen Test

Nakagawa

Maeda

Murai

2002

Clin Vaccine Immunol

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“…A dose of 13.81 EU/mL/kg of the 2 nd International Standard for bacterial endotoxins was identified as that which produced a 0.5ºC rise in temperature (Fig. 1); this value, although variable according to the strain of the animals used, is recommended as a criterion for positive responses (9,17). Despite its shortcomings, the test is recommended by the Pharmacopoeias (2,3,20), and it is important for the validation of new in vitro assays under development (4,7,16).…”

Section: Discussionmentioning

confidence: 99%

“…However, the test can give falsenegative and false-positive results and, being a test for Gramnegative LPS, it does not detect Gram-positive exotoxins, viruses and fungi (7,13,16). In the LAL test, the LPS-induced reaction can be measured using various approaches, which were adopted together with the rabbit pyrogen test in the collaborative studies that established the 1 st and 2 nd International Standard for bacterial endotoxins (15,17). However, despite the specification of the LAL test for pharmaceutical products, there remain a number of complex preparations, such as biologicals, without specifications or that cannot be tested by the LAL test (6).…”

Section: Introductionmentioning

confidence: 99%

Comparative evaluation of pyrogens tests in pharmaceutical products

Silveira¹,

Andrade²,

Schmidt³

et al. 2004

Braz. J. Microbiol.

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A comparison of methodologies for detection of pyrogens in pharmaceutical products was performed. The rabbit pyrogen test was optimized and the dose-response curve was obtained for the 2 nd International Standard for bacterial endotoxins, establishing 13.81 EU/mL/kg as the concentration of endotoxin necessary to induce a temperature rise of 0.5ºC. The 0.5ºC cut-off was shown to give results that were more compatible with the pyrogenic doses for humans. The Limulus amoebocyte lysate test (LAL) was standardized with gel-clot and chromogenic endpoints, and used for the comparative evaluation of pharmaceutical products showing good agreement. The use of β-glucan-reactive and non-reactive LAL reagents identified some products with falsepositive results. The interference test was carried out and the specifications validated for some new products as the maximum valid dilution. The results emphasized the importance and limitations of the assays recommended for the evaluation of purity and quality control of parenteral medicinal products, improving the existing methodologies in the context of reduction and replacement in the use of animal models.

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“…100 pg of this LPS are defined as 1 EU (Poole and Mussett, 1989). LTA from Staphylococcus aureus was isolated and prepared in-house as described previously (Morath et al, 2001).…”

Section: Pyrogensmentioning

confidence: 99%

In vitro pyrogen test for toxic or immunomodulatory drugs

Daneshian

Guenther

Wendel

et al. 2006

Journal of Immunological Methods

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Pyrogenic contaminations of some classes of injectable drugs, e.g , tox ic or immunomodulatory as well as false-positive drugs, represent a major risk which cannot yet be excluded due to the limitations of current tests . Here we describe a modification of the In vitro Pyrogen Test termed AWIPT (Adsorb, Wash, In vitro Pyrogen Test), which addresses this problem by introducing a preincubation step in which pyrogenic contaminations in the test sample are adsorbed to albumin-coated beads. After rinsing, the beads are incubated with human whole blood and the release of the endogenous pyrogen interleukin-I~is measured as a marker of pyrogenic activity. Intentional contaminations with lipopolysaccharide were retrieved from the chemotherapeutic agents paclitaxel, cisplatin and liposomal daunorubicin, the antibiotic gentamicin, the antifungal agent liposomal amphotericin B, and the cort icosteroid prednisolone at lower dilut ions than in the standard in vitro pyrogen test. This represents a promising new approach for the detection of pyrogenic contamination in drugs or in drugs containing interfering additives and should lead to improved safety levels .

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The International Standard for Endotoxin: evaluation in an international collaborative study

Cited by 32 publications

References 4 publications

Evaluation of the In Vitro Pyrogen Test System Based on Proinflammatory Cytokine Release from Human Monocytes: Comparison with a Human Whole Blood Culture Test System and with the Rabbit Pyrogen Test

Evaluation of the In Vitro Pyrogen Test System Based on Proinflammatory Cytokine Release from Human Monocytes: Comparison with a Human Whole Blood Culture Test System and with the Rabbit Pyrogen Test

Comparative evaluation of pyrogens tests in pharmaceutical products

In vitro pyrogen test for toxic or immunomodulatory drugs

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