Immunological strategy for detecting the pro-inflammatory cytokine TNF-alpha in salmonids
Electron. j. biotechnol
;
15(5): 21-21, Sept. 2012. ilus, tab
Artículo
en Inglés
| LILACS
| ID: lil-657680
ABSTRACT
Background:
Tumour necrosis factor-alpha (TNF-alpha) is a pro-inflammatory cytokine which exerts a variety of immunological functions in vertebrates. TNF-alpha has been identified and cloned in a number of teleost fish species; nevertheless, the functions displayed by this cytokine in fishes remain poorly understood, given that the low sequence identity compared to their mammalian counterpart, limit fish TNF-alpha detection using mammalian antibodies. Then, for fish immune response characterization is fundamental the production of specific fish anti-TNF-alpha antibody.Results:
We have developed a monoespecific antibody against the pro-inflammatory molecule TNF-alpha of salmonid fish. TNF-alpha epitope region was identified and characterized using bioinformatic tools. The epitope sequence was chemically synthesized using Fmoc strategy, analyzed by RP-HPLC and its molecular weight confirmed by mass spectrometry. The synthetic peptide was used to immunize mice and antibodies from ascitic fluid were purified. The resulting antibody was used for molecular and histochemical detection in gut samples from salmonid fishes treated with different food. By ELISA, we detected a differential expression of TNF-alpha, the western blot analysis shows recognition of the whole TNF molecule and by immunohistochemistry TNF-alpha positive cells were observed.Conclusions:
We provide an immunological tool, validated through classical immunological assays, which can be a useful tool for characterizing fish TNF-alpha function.
Texto completo:
Disponible
Índice:
LILACS (Américas)
Asunto principal:
Salmonidae
/
Factor de Necrosis Tumoral alfa
/
Mediadores de Inflamación
Tipo de estudio:
Estudio pronóstico
Límite:
Animales
Idioma:
Inglés
Revista:
Electron. j. biotechnol
Asunto de la revista:
Biotecnologia
Año:
2012
Tipo del documento:
Artículo
/
Documento de proyecto
País de afiliación:
Chile
Institución/País de afiliación:
Pontificia Universidad Católica de Valparaíso/CL
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