RESUMO
The need for alternatives to animal use in pyrogen testing has been driven by the Three Rs concept. This has resulted in the inclusion of the monocyte activation test (MAT) in the European Pharmacopoeia, 2010. However, some technical and regulatory obstacles must be overcome to ensure the effective implementation of the MAT by the industry, especially for the testing of biological products. The yellow fever (YF) vaccine (17DD-YFV) was chosen for evaluation in this study, in view of: a) the 2016-2018 outbreak of YF in Brazil; b) the increase in demand for 17DD-YFV doses; c) the complex production process with live attenuated virus; d) the presence of possible test interference factors, such as residual process components (e.g. ovalbumin); and e) the need for the investigation of other pyrogens that are not detectable by the methods prescribed in the YF vaccine monograph. The product-specific testing was carried out by using cryopreserved and fresh whole blood, and IL-6 and IL-1ß levels were used as the marker readouts. After assessing the applicability of the MAT on a 1:10 dilution of 17DD-YFV, endotoxin and non-endotoxin pyrogens were quantified in spiked batches, by using the lipopolysaccharide and lipoteichoic acid standards, respectively. The quantitative analysis demonstrated the correlation between the MAT and the Limulus amoebocyte lysate (LAL) assays, with respect to the limits of endotoxin recovery in spiked batches and the detection of no pyrogenic contamination in commercial batches of 17DD-YFV. The data demonstrated the applicability of the MAT for 17DD-YFV pyrogen testing, and as an alternative method that can contribute to biological quality control studies.
Assuntos
Alternativas aos Testes com Animais , Monócitos/efeitos dos fármacos , Pirogênios/análise , Controle de Qualidade , Vacina contra Febre Amarela/normas , Animais , Humanos , Interleucina-1beta/sangue , Interleucina-6/sangue , Teste do Limulus , Lipopolissacarídeos/análise , Monócitos/imunologiaRESUMO
Borrelia burgdorferi é uma espiroqueta transmitida por carrapatos, de ampla distribuição geográfica, que acomete o homeme animais domésticos, tendo os animais silvestres como reservatórios. O presente estudo teve como objetivo relatar aocorrência de eqüinos soropositivos para B. burgdorferi, por meio de inquérito soroepidemiológico utilizando o ensaio deimunoadsorção enzimática (ELISA) indireto. Procedeu-se a análise sorológica de 747 soros de eqüinos, onde 431 pertenciamao município de Três Rios e 316 ao de Vassouras, ambos do estado do Rio de Janeiro. Um total de 212 (28,4%) animais forampositivos ao ensaio. O resultado observado foi similar aos descritos para animais no Brasil e Estados Unidos da América,tendo sido verificado que os animais apresentaram anticorpos homólogos anti-Borrelia burgdorferi stricto sensu, corroborandocom a ocorrência de Borrelia sp. em eqüinos no estado do Rio de Janeiro
Lyme Borreliosis is a widely distributed spirochetosis hosted by wild animals, which transmitted to domestic animals as wellas man by ticks. The present study aims to observe the occurrence level of positive tested equines to Borrelia burgdorferi, on twomunicipalities of Rio de Janeiro State, Brazil, by seroepidemiologic inquiry using the indirect Enzime-Linked ImmunosorbentAssay. A serologic analysis was performed on 747 equines serum specimens, which 431 were from Três Rios and 316 werefrom Vassouras municipality. For this test, 212 (28.4%) animals were positive. The results observed were similar to thosedescribed for animals in Brazil and United States of America, confirming that the animals of the studied municipalities showedanti Borrelia burgdorferi homologs antibodies