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In vitro human immune reactivity of fast protein liquid chromatography fractionated Paracoccidioides brasiliensis soluble antigens.
Diniz, S N; Cisalpino, P S; Koury, M C; Andrade, G M; Nogueira, M G; Goes, A M.
Afiliação
  • Diniz SN; Departamento de Bioquímica e Imunologia, Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, Minas Gerais, 30 161-970, Brazil.
Microbes Infect ; 1(5): 353-60, 1999 Apr.
Article em En | MEDLINE | ID: mdl-10602668
ABSTRACT
Soluble antigens of Paracoccidioides brasiliensis yeast cells (PbAg) were fractionated in a fast protein liquid chromatography (FPLC) system, using Q-Sepharose anion-exchange resin, in order to characterize antigenic fractions that could elicit cell reactivity and antibody recognition in human paracoccidioidomycosis (PCM). PbAg fractions were eluted by 20 mM Tris-HCl solution (pH 9.6) with an increasing gradient up to 1 M NaCl. The FPLC system was able to resolve 7 fractions, enumerated from 0 to VI, according to the elution on the NaCl gradient. The analysis of each fraction on SDS-PAGE showed that fractions 0 to V were constituted by multiple protein bands with molecular mass ranging from 18 to 114 kDa. Large amounts of nucleic acids were evidenced in fraction VI, as revealed by agarose gel stained with ethidium bromide. Sera from PCM patients presenting different clinical forms contained antibodies that recognized antigens in all fractions with the exception of fraction VI as detected by ELISA. Further studies were designed to investigate the capacity of these fractions to induce cell proliferation. It was demonstrated that fractions III and V (200 and 450 mM NaCl, respectively) stimulated a significant proliferative response of peripheral blood mononuclear cells, while fraction 0 induced the lowest proliferative response among patients with PCM, in either acute, acute treated, or chronic forms.
Assuntos
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Base de dados: MEDLINE Assunto principal: Paracoccidioides / Paracoccidioidomicose / Antígenos de Fungos Limite: Adolescent / Adult / Child / Child, preschool / Humans / Middle aged País/Região como assunto: America do sul / Brasil Idioma: En Revista: Microbes Infect Assunto da revista: ALERGIA E IMUNOLOGIA / MICROBIOLOGIA Ano de publicação: 1999 Tipo de documento: Article País de afiliação: Brasil
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Base de dados: MEDLINE Assunto principal: Paracoccidioides / Paracoccidioidomicose / Antígenos de Fungos Limite: Adolescent / Adult / Child / Child, preschool / Humans / Middle aged País/Região como assunto: America do sul / Brasil Idioma: En Revista: Microbes Infect Assunto da revista: ALERGIA E IMUNOLOGIA / MICROBIOLOGIA Ano de publicação: 1999 Tipo de documento: Article País de afiliação: Brasil