Novel Pyrogen Tests Based on the Human Fever Reaction

Hartung, Thomas; Aaberge, Ingeborg S.; Berthold, S. Megan; Carlin, Gunnar; Charton, Emmanuelle; Coecke, Sandra; Fennrich, Stefan; Fischer, Matthias; Gommer, Martin; Halder, Marlies; Hasløv, K; Jahnke, M.; Montag-Lessing, Thomas; Poole, Stephen; Schechtman, Leonard M.; Wendel, Albrecht; Werner-Felmayer, Gabriele

doi:10.1177/026119290102900203

Cited by 59 publications

(14 citation statements)

References 123 publications

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“…The in-house developed ELISA [22] for the detection of IL-1 β requires minimal incubation time and is based on Sandwich method of antigen detection [24, 25]. The advantage of cryopreserved lymphocytes over fresh human whole blood and fresh lymphocytes is the off-shelf availability of screened, pathogen-free, and allergen-free system for IL-1 β detection.…”

Section: Discussionmentioning

confidence: 99%

Analysis of IL-1βRelease from Cryopreserved Pooled Lymphocytes in Response to Lipopolysaccharide and Lipoteichoic Acid

Nair

Geetha

Mohanan

2013

BioMed Research International

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Pyrogens are heterogeneous group of fever-inducing substances derived from Gram-positive and Gram-negative bacteria, fungi, and viruses. They incite immune response by producing endogenous pyrogens such as prostaglandins and other proinflammatory cytokines like IL-1β, IL-6, and TNF-α. The present study was to analyze the influence of cryopreservation in IL-1β release, a marker for inflammatory response from human lymphocytes, in response to exogenous pyrogenic stimulants. Lymphocytes isolated from pooled blood of multiple healthy individuals were cryopreserved in DMSO and glycerol for periods of 7, 14, 30, and 60 days and were challenged with LPS and LTA in vitro. The inflammatory cytokine, IL-1β release, was measured by ELISA method. It was observed that the release of IL-1β increases instantaneously after the initiation of incubation and reaches a maximum at 3 to 5 hours and then gradually decreases and gets stabilized for both pyrogens. Moreover it was also observed that the effect of cryoprotectants, DMSO (10%) and glycerol (10%), showed almost similar results for short-term storage, but DMSO-preserved lymphocytes yielded a better viability for long-term storage. Thus, the isolated cryopreserved lymphocytes system can be a promising approach for the total replacement/alteration to animal experimentation for pyrogenicity evaluation.

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Section: Discussionmentioning

confidence: 99%

Analysis of IL-1βRelease from Cryopreserved Pooled Lymphocytes in Response to Lipopolysaccharide and Lipoteichoic Acid

Nair

Geetha

Mohanan

2013

BioMed Research International

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“…The induction of proinflammatory cytokine by the extracts was assayed with an in vitro pyrogen test system using human monocytic cells, MM6-CA8, which respond to various pyrogens with high sensitivity. 24,25 MM6-CA8 cells were maintained in RPMI 1640 medium (Gibco, NY) containing fetal bovine serum (10%), glutamine (2 mM), non-essential amino acids (0.1 mM), sodium pyruvate (1 mM), and bovine insulin (9 lg/mL). The cells were primed with 10 ng/mL of calcitriol (1a, 25-dihydroxyvitamin D 3 , Wako Pure Chemical Industries, Osaka, Japan).…”

Section: Release Of Proinflammatory Cytokines From Human Monocytic Cellsmentioning

confidence: 99%

A biological study establishing the endotoxin limit of biomaterials for bone regeneration in cranial and femoral implantation of rats

et al. 2016

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The purpose of this study was to accurately quantify the risk of endotoxin contamination in biomaterials for bone regeneration in order to establish the acceptable endotoxin limit. Collagen sheets containing varying amounts of purified endotoxin from Escherichia coli and dried, heat-killed E. coli or Staphylococcus aureus cells were implanted into cranial or femoral defects in rats. These defects were artificially prepared to a size of 5 × 5 mm or a diameter of 1 mm, respectively. The degree of osteoanagenesis was assessed by soft X-ray radiography and histopathology at 1 and 4 weeks after implantation. The collagen sheet containing the dried E. coli cells showed a dose-dependent delay in cranial and/or femoral osteoanagenesis at endotoxin activities of more than 33.6 EU/mg, at which no inflammatory response was observed. In contrast, no such observation occurred with the collagen sheet containing S. aureus cells. These results suggest that endotoxins may affect the process of osteoanagenesis. Additionally, the no-observed-adverse-effect level was 9.6 EU/mg, corresponding to 255 EU/kg body weight in rats. Interestingly, no delay in osteoanagenesis was induced by the implantation of collagen sheets containing purified endotoxin at any dose tested. This suggested that pure endotoxin implanted into tissues having poor circulation of bodily fluids without bleeding may not be recognized as a foreign substance and may not induce a significant biological response. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1514-1524, 2017.

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“…O RPT tem sido tradicionalmente recomendado pelas farmacopeias para avaliar contaminantes pirogênicos em parenterais, dispositivos médicos e produtos biológicos (Welch et al, 1943, Du et al, 2011. Entretanto, dentre os fatores limitantes deste teste estão o manejo dos animais, a confiabilidade dos ensaios (resultados falsopositivo ou falso-negativos) (Grant, 1950, Hartung, 2015, o fato de não ser quantitativo (teste limite), não utilizar controle positivo, não possuir diferencial de sensibilidade aos pirogênios devido a extrapolação interespécies (coelho/homem) (ICCVAM, 2008), restritiva aplicabilidade a uma gama de produtos farmacêuticos modernos (radiofármacos, analgésicos, biofármacos, drogas imunomodulatórias e drogas contra câncer) que não podem ser testados em coelhos devido aos efeitos interferentes (Hartung et al, 2001;Hartung, 2015) por influenciarem na regulação da temperatura ou por causarem reações. Além disso, o RPT não pode ser utilizado para testes de preparações celulares (Hartung et al, 2001) (Tabela 1).…”

Section: Vantagens Do Mat Como Método Alternativounclassified

“…IgG intravenosa) (Huszár et al, 2002) e suspensões oleosas ou detergentes (Hasiwa et al, 2013). Além disso, o ensaio RPT não pode ser usado para as preparações celulares, tais como elementos figurados e células tronco (Hartung et al, 2001) Drogas que interferem com o sistema de coagulação do teste LAL, através da inibição gerando resultado falsonegativo (ligação de cátions divalentes-EDTA/ citrato e inibidores da protease) ou pela indução com resultado falso-positivo (alto teor de proteína, as proteases), não podem ser testadas por este sistema (Dawson, 2005). Polissacarídeos podem ser contaminantes obtidos do processo de purificação e derivados de materiais de filtro de celulose, podendo originar resultados falso-positivos (Henne et al, 1984).…”

Section: Avaliação De Pirogênios Em Produtos Biológicosunclassified

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Métodos alternativos ao uso de animais para a detecção de pirogênio: oportunidades e desafios no controle da qualidade de produtos biológicos.

Navega

Silva

Presgrave

et al. 2016

AVS

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A necessidade de alternativas ao uso de animais no teste de pirogênio foi guiada pelo principio dos 3R’s, culminando no desenvolvimento e aceitação pela Farmacopeia Europeia do método alternativo in vitro, o Teste de Ativação de Monócitos (Monocyte Activation Test- MAT, 2.6.30/ 2010). O MAT utiliza como matriz, fontes de monócitos humanos (sangue total, PBMC, linhagem), respeitando o conceito dos 3R’s e excluindo riscos inerentes à extrapolação inter-espécies. É um método promissor e eficiente, que detecta um amplo espectro de pirogênios e supri limitações dos testes atuais preconizados pelas farmacopeias, o teste in vitro Teste de Endotoxina Bacteriana e o Teste de Pirogênio em Coelhos. Apesar das vantagens abordadas, alguns obstáculos técnico-científicos e regulatórios devem ser transpostos para a implantação efetiva do MAT na rotina industrial, em especial de produtos biológicos.

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Novel Pyrogen Tests Based on the Human Fever Reaction

Cited by 59 publications

References 123 publications

Analysis of IL-1βRelease from Cryopreserved Pooled Lymphocytes in Response to Lipopolysaccharide and Lipoteichoic Acid

Analysis of IL-1βRelease from Cryopreserved Pooled Lymphocytes in Response to Lipopolysaccharide and Lipoteichoic Acid

A biological study establishing the endotoxin limit of biomaterials for bone regeneration in cranial and femoral implantation of rats

Métodos alternativos ao uso de animais para a detecção de pirogênio: oportunidades e desafios no controle da qualidade de produtos biológicos.

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