title: [Marek's disease virus can infect chicken brain microglia and promote the transcription of toll-like receptor 15 and 1LB genes].
abstract: Microglial cells were purified from a mixed neuroglia culture prepared from the neonatal chicken brain in vitro, and were infected with the vvMDV YL040920 isolate and an attenuated MDV vaccine strain CVI988/Rispens, respectively. The presence of cytopathic effect (CPE) was examined daily, and the MEQ expression in MDV-infected microglia was detected by immunohistochemistry assay. DNA replication of the MDV meq gene and transcription of the gB gene were determined by real-time quantitative PCR (qPCR) and qRT-PCR, respectively. The transcripts of Toll-like receptor (TLR) mRNA in microglia post MDV infection were quantified by qRT-PCR. The results of this study showed that both vvMDV YL040920 and attenuated vaccine strain CVI988/Rispens could infect microglia and produce characteristic CPE with plaque formation. The plaques were formed due to cells shedding at multi-sites, then quickly expanded and integrated. Furthermore, the MEQ protein was detected in nuclei of YL040920 and CVI988/ Rispens-infected microglia, and MDV meq DNA replication and gB gene transcription in MDV-infected microglia were also confirmed. Although both MDV DNA copies and gB transcripts were increased in the virus-infected microglia, the higher viral DNA load and gB transcript were observed for CVI988/Rispens than for YL040920 in vitro (P < or = 0.05/0.001). The transcriptions of TLR15 and TLR1LB gene were found to be up-regulated in microglia following MDV infection in vitro. Purified microglia infected with YL040920 was observed increased TLR15 and TLR1LB transcripts as early as 1 day post infection (dpi), and reached its peak level at 3 dpi, then decreased mildly at 5 dpi. For CVI988/Rispens, it induced an increase of TLR15 transcript as early as 1 dpi, and rose rapidly at 3 dpi, and then decreased slightly at 5 dpi. At the same time, CVI988/Rispens induced the increase of chTLR1LB transcript at 3 dpi and decreased at 5 dpi. By comparing the TLRs transcription between YL040920 and CVI988/Rispens-infected microglia, it was suggested that vvMDV YL040920 might induce more TLR15 transcript than the attenuated vaccine strain CVI988/Rispens (P < or = 0.01/0.001), while CVI988/Rispens induced more TLR1LB transcript than YL040920 (P < or = 0.001).
authors.affiliation: Guangxi University, Nanning 530005. eyangql@sina.com
authors.forename: Qing-li
authors.initials: QL
authors.lastname: Yang
authors.forename: Hao
authors.initials: H
authors.lastname: Chen
authors.forename: Ping
authors.initials: P
authors.lastname: Wei
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000821:virology; D002645:Chickens; D015870:Gene Expression; D008381:Herpesvirus 2, Gallid / Q000235:genetics / Q000502:physiology*; D008380:Marek Disease / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D017628:Microglia / Q000378:metabolism* / Q000821:virology; D011201:Poultry Diseases / Q000235:genetics* / Q000378:metabolism / Q000821:virology; D051194:Toll-Like Receptor 1 / Q000235:genetics* / Q000378:metabolism; D051193:Toll-Like Receptors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D051194:Toll-Like Receptor 1; D051193:Toll-Like Receptors