title: Culture of human amniotic cells: a system to study interferon production.
abstract: This study investigated whether primary culture of human amniotic membrane cells (PCHAM) could be used as an in vitro model system for the study of interferon (IFN) production. PCHAM cells infected with Newcastle disease virus (NDV) produced the two antigenic types of IFN, previously shown in a amniotic membrane cells (HAM) system. PCHAM IFN was detected as early as 2 h after NDV infection and was composed by two antigenically distinct fractions, one neutralized with anti-HuIFN beta antibody and another that is not related to IFN beta, -alpha and -gamma. These fractions correspond respectively to 80 and 20 per cent of the IFN produced 4 h after virus induction, 55 and 45 per cent of the IFN produced from 4 to 12 h and 67 and 33 per cent of the IFN produced 12 h after virus induction. A cDNA library, established from PCHAM with or without NDV infection, was screened for IFN alpha and -beta using specific primers. The PCR product, amplified by IFN beta primers, was cloned, sequenced and expressed in Escherichia coli M15. The sequences of several cloned cDNAs were identical to HuIFN beta gene and the antiviral activity of the expressed protein was neutralized only by antiHuIFN-beta antibody. The other IFN fraction not neutralized by polyclonal antibodies anti-IFN beta, -alpha and -gamma is now being studied.
authors.affiliation: Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: A F
authors.initials: AF
authors.lastname: Carvalho
authors.forename: J R
authors.initials: JR
authors.lastname: Santos
authors.forename: R
authors.initials: R
authors.lastname: Gentz
authors.forename: C A
authors.initials: CA
authors.lastname: Bonjardim
authors.forename: R R
authors.initials: RR
authors.lastname: Golgher
authors.forename: P C
authors.initials: PC
authors.lastname: Ferreira
authors.forename: E G
authors.initials: EG
authors.lastname: Kroon
mesh_terms: D000650:Amnion / Q000166:cytology* / Q000276:immunology / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D018920:Coculture Techniques; D017931:DNA Primers / Q000235:genetics; D005260:Female; D006801:Humans; D016899:Interferon-beta / Q000096:biosynthesis / Q000235:genetics; D007372:Interferons / Q000096:biosynthesis* / Q000235:genetics; D007700:Kinetics; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D016899:Interferon-beta; D007372:Interferons