Thermolabile and calcium-dependent serum factor interferes with polymerized actin, and impairs anti-actin antibody detection

Thermolabile and calcium-dependent serum factor interferes with polymerized actin, and impairs anti-actin antibody detection

Autor Cancado, ELR Google Scholar
Abrantes-Lemos, C. P. Google Scholar
Vilas-Boas, L. S. Google Scholar
Novo, N. F. Google Scholar
Carrilho, F. J. Google Scholar
Laudanna, A. A. Google Scholar
Instituição Universidade de São Paulo (USP)
Universidade Federal de São Paulo (UNIFESP)
Resumo The detection of anti-actin (AAA) by immunofluorescence is hindered by the presence of a serum factor. To better understand how it interferes with AAA detection, we tested sera from 20 patients with autoimmune hepatitis, and from 21 healthy adults, diluted 1:10 and prepared as follows: (A) diluted with PBS; (B) inactivated at 56 degreesC, and diluted with PBS; (C) diluted with 34 mM EDTA/PBS; (D) heated and diluted with EDTA/PBS. To reveal AAA, a fluorescein-labelled anti-human IgG was used in the process of indirect immunofluorescence. in a parallel assay, the substrate, acetone-fixed human fibroblasts, was preincubated with-sera prepared as if it were to identify AAA, but instead, a rhodamine-phalloidin was used to identify F-actin, by direct immunofluorescence. All sera from patients were reactive to AAA when heat-inactivated and/or calcium-chelated, and 60% of them when diluted with unmodified sera (P = 0.004). F-actin continued to be present after preincubation with heat-inactivated or calcium-chelated sera from patients and healthy controls, and in 41.5% of reactions with unmodified serum (P = 0.0000001). the heat inactivation and the calcium chelation were both efficient procedures for maintaining the microfilament structure intact after serum incubation and, therefore, for identifying AAA. (C) 2001 Academic Press.
Palavra-chave anti-smooth antibody
autoantibodies
autoimmune hepatitis
microfilaments
gelsolin
Idioma Inglês
Data de publicação 2001-11-01
Publicado em Journal of Autoimmunity. London: Academic Press Ltd, v. 17, n. 3, p. 223-228, 2001.
ISSN 0896-8411 (Sherpa/Romeo, fator de impacto)
Publicador Academic Press Ltd
Extensão 223-228
Fonte http://dx.doi.org/10.1006/jaut.2001.0540
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000172427400007
Endereço permanente http://repositorio.unifesp.br/handle/11600/26652

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