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An in vitro experimental model of neuroinflammation: the induction of interleukin‐6 in murine astrocytes infected with Theiler’s murine encephalomyelitis virus, and its inhibition by oestrogenic receptor modulators
Author(s) -
Rubio Nazario,
Cerciat Marie,
Unkila Mikko,
GarciaSegura Luis M.,
Arevalo MariaAngeles
Publication year - 2011
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1111/j.1365-2567.2011.03448.x
Subject(s) - biology , neuroinflammation , microbiology and biotechnology , in vitro , virology , interleukin , microglia , astrocyte , recombinant dna , tumor necrosis factor alpha , receptor , cytokine , immunology , inflammation , central nervous system , gene , biochemistry , neuroscience
Summary This paper describes an experimental model of neuroinflammation based on the production of interleukin‐6 (IL‐6) by neural glial cells infected with Theiler’s murine encephalomyelitis virus (TMEV). Production of IL‐6 mRNA in mock‐infected and TMEV‐infected SJL/J murine astrocytes was examined using the Affymetrix murine genome U74v2 DNA microarray. The IL‐6 mRNA from infected cells showed an eightfold increase in hybridization to a sequence encoding IL‐6 located on chromosome number 5. Quantitative real‐time reverse transcription PCR (qPCR) was used to study the regulation of IL‐6 expression. The presence of IL‐6 in the supernatants of TMEV‐infected astrocyte cultures was quantified by ELISA and found to be weaker than in cultures of infected macrophages. The IL‐6 was induced by whole TMEV virions, but not by Ad.βGal adenovirus, purified TMEV capsid proteins, or UV‐inactivated virus. Two recombinant inflammatory cytokines, IL‐1α and tumour necrosis factor‐α were also found to be potent inducers of IL‐6. The secreted IL‐6 was biologically active because it fully supported B9 hybridoma proliferation in a [ 3 H]thymidine incorporation bioassay. The cerebrospinal fluid of infected mice contained IL‐6 during the acute encephalitis phase, peaking at days 2–4 post‐infection. Finally, this in vitro neuroinflammation model was fully inhibited, as demonstrated by ELISA and qPCR, by five selective oestrogen receptor modulators.