PMID- 21325477
OWN - NLM
STAT- MEDLINE
DCOM- 20110712
LR  - 20231213
IS  - 1465-2099 (Electronic)
IS  - 0022-1317 (Linking)
VI  - 92
IP  - Pt 6
DP  - 2011 Jun
TI  - Single cell detection of latent cytomegalovirus reactivation in host tissue.
PG  - 1279-1291
LID - 10.1099/vir.0.029827-0 [doi]
AB  - The molecular mechanisms leading to reactivation of latent cytomegalovirus are 
      not well understood. To study reactivation, the few cells in an organ tissue that 
      give rise to reactivated virus need to be identified, ideally at the earliest 
      possible time point in the process. To this end, mouse cytomegalovirus (MCMV) 
      reporter mutants were designed to simultaneously express the red fluorescent 
      protein mCherry and the secreted Gaussia luciferase (Gluc). Whereas Gluc can 
      serve to assess infection at the level of individual mice by measuring 
      luminescence in blood samples or by in vivo imaging, mCherry fluorescence offers 
      the advatage of detection of infection at the single cell level. To visualize 
      cells in which MCMV was being reactivated, precision-cut lung slices (PCLS) that 
      preserve tissue microanatomy were prepared from the lungs of latently infected 
      mice. By day 3 of cultivation of the PCLS, reactivation was revealed by Gluc 
      expression, preceding the detection of infectious virus by approximately 4 days. 
      Reactivation events in PCLS could be identified when they were still confined to 
      single cells. Notably, using fractalkine receptor-GFP reporter mice, we never 
      observed reactivation originating from CX3CR1(+) monocytes or pulmonary dendritic 
      cells derived therefrom. Furthermore, latent viral genome in the lungs was not 
      enriched in sorted bone-marrow-derived cells expressing CD11b. Taken together, 
      these complementary approaches suggest that CD11b(+) and CX3CR1(+) subsets of the 
      myeloid differentiation lineage are not the main reservoirs and cellular sites of 
      MCMV latency and reactivation in the lungs.
FAU - Marquardt, Anja
AU  - Marquardt A
AD  - Institute of Virology, Hannover Medical School, Hannover, Germany.
FAU - Halle, Stephan
AU  - Halle S
AD  - Institute of Immunology, Hannover Medical School, Hannover, Germany.
FAU - Seckert, Christof K
AU  - Seckert CK
AD  - Institute for Virology, University Medical Center of the Johannes 
      Gutenberg-University, Mainz, Germany.
FAU - Lemmermann, Niels A W
AU  - Lemmermann NAW
AD  - Institute for Virology, University Medical Center of the Johannes 
      Gutenberg-University, Mainz, Germany.
FAU - Veres, Tibor Z
AU  - Veres TZ
AD  - Department of Immunology, Allergology and Immunotoxicology, Fraunhofer Institute 
      of Toxicology and Experimental Medicine, Hannover, Germany.
FAU - Braun, Armin
AU  - Braun A
AD  - Department of Immunology, Allergology and Immunotoxicology, Fraunhofer Institute 
      of Toxicology and Experimental Medicine, Hannover, Germany.
FAU - Maus, Ulrich A
AU  - Maus UA
AD  - Department of Experimental Pneumology, Hannover Medical School, Hannover, 
      Germany.
FAU - Forster, Reinhold
AU  - Forster R
AD  - Institute of Immunology, Hannover Medical School, Hannover, Germany.
FAU - Reddehase, Matthias J
AU  - Reddehase MJ
AD  - Institute for Virology, University Medical Center of the Johannes 
      Gutenberg-University, Mainz, Germany.
FAU - Messerle, Martin
AU  - Messerle M
AD  - Institute of Virology, Hannover Medical School, Hannover, Germany.
FAU - Busche, Andreas
AU  - Busche A
AD  - Institute of Virology, Hannover Medical School, Hannover, Germany.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110216
PL  - England
TA  - J Gen Virol
JT  - The Journal of general virology
JID - 0077340
RN  - 0 (Luminescent Proteins)
RN  - EC 1.13.12.- (Luciferases)
SB  - IM
MH  - Animals
MH  - Cytomegalovirus/genetics/physiology
MH  - Cytomegalovirus Infections/*virology
MH  - Disease Models, Animal
MH  - Female
MH  - Gene Expression
MH  - Genes, Reporter
MH  - Herpesviridae Infections/virology
MH  - Host-Pathogen Interactions
MH  - Humans
MH  - Luciferases/genetics/metabolism
MH  - Luminescent Proteins/genetics/metabolism
MH  - Lung/cytology/virology
MH  - Male
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Muromegalovirus/genetics/isolation & purification/*physiology
MH  - Single-Cell Analysis/*methods
MH  - *Virus Activation
MH  - *Virus Latency
MH  - Red Fluorescent Protein
EDAT- 2011/02/18 06:00
MHDA- 2011/07/13 06:00
CRDT- 2011/02/18 06:00
PHST- 2011/02/18 06:00 [entrez]
PHST- 2011/02/18 06:00 [pubmed]
PHST- 2011/07/13 06:00 [medline]
AID - 10.1099/vir.0.029827-0 [doi]
PST - ppublish
SO  - J Gen Virol. 2011 Jun;92(Pt 6):1279-1291. doi: 10.1099/vir.0.029827-0. Epub 2011 
      Feb 16.