title: 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.
abstract: 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.
authors.affiliation: Instituto Cajal. C.S.I.C, Madrid, Spain. nazario@cajal.csic.es
authors.forename: Nazario
authors.initials: N
authors.lastname: Rubio
authors.forename: Marie
authors.initials: M
authors.lastname: Cerciat
authors.forename: Mikko
authors.initials: M
authors.lastname: Unkila
authors.forename: Luis M
authors.initials: LM
authors.lastname: Garcia-Segura
authors.forename: Maria-Angeles
authors.initials: MA
authors.lastname: Arevalo
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000187:drug effects* / Q000821:virology*; D018188:Cardiovirus Infections / Q000276:immunology* / Q000503:physiopathology; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D020847:Estrogen Receptor Modulators / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D007249:Inflammation / Q000188:drug therapy; D020382:Interleukin-18 / Q000378:metabolism; D015850:Interleukin-6 / Q000134:cerebrospinal fluid / Q000276:immunology* / Q000378:metabolism; D008264:Macrophages / Q000821:virology; D051379:Mice; D018448:Models, Immunological; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017945:Theilovirus / Q000276:immunology
chemical_list: D020847:Estrogen Receptor Modulators; D020382:Interleukin-18; D015850:Interleukin-6