PMID- 26984724
OWN - NLM
STAT- MEDLINE
DCOM- 20170426
LR  - 20181113
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 90
IP  - 10
DP  - 2016 May 15
TI  - Porcine Reproductive and Respiratory Syndrome Virus Utilizes Nanotubes for 
      Intercellular Spread.
PG  - 5163-5175
LID - 10.1128/JVI.00036-16 [doi]
AB  - Intercellular nanotube connections have been identified as an alternative pathway 
      for cellular spreading of certain viruses. In cells infected with porcine 
      reproductive and respiratory syndrome virus (PRRSV), nanotubes were observed 
      connecting two distant cells with contiguous membranes, with the core infectious 
      viral machinery (viral RNA, certain replicases, and certain structural proteins) 
      present in/on the intercellular nanotubes. Live-cell movies tracked the 
      intercellular transport of a recombinant PRRSV that expressed green fluorescent 
      protein (GFP)-tagged nsp2. In MARC-145 cells expressing PRRSV receptors, GFP-nsp2 
      moved from one cell to another through nanotubes in the presence of 
      virus-neutralizing antibodies. Intercellular transport of viral proteins did not 
      require the PRRSV receptor as it was observed in receptor-negative HEK-293T cells 
      after transfection with an infectious clone of GFP-PRRSV. In addition, GFP-nsp2 
      was detected in HEK-293T cells cocultured with recombinant PRRSV-infected 
      MARC-145 cells. The intercellular nanotubes contained filamentous actin (F-actin) 
      with myosin-associated motor proteins. The F-actin and myosin IIA were identified 
      as coprecipitates with PRRSV nsp1beta, nsp2, nsp2TF, nsp4, nsp7-nsp8, GP5, and N 
      proteins. Drugs inhibiting actin polymerization or myosin IIA activation 
      prevented nanotube formation and viral clusters in virus-infected cells. These 
      data lead us to propose that PRRSV utilizes the host cell cytoskeletal machinery 
      inside nanotubes for efficient cell-to-cell spread. This form of virus transport 
      represents an alternative pathway for virus spread, which is resistant to the 
      host humoral immune response. IMPORTANCE: Extracellular virus particles transmit 
      infection between organisms, but within infected hosts intercellular infection 
      can be spread by additional mechanisms. In this study, we describe an alternative 
      pathway for intercellular transmission of PRRSV in which the virus uses nanotube 
      connections to transport infectious viral RNA, certain replicases, and certain 
      structural proteins to neighboring cells. This process involves interaction of 
      viral proteins with cytoskeletal proteins that form the nanotube connections. 
      Intercellular viral spread through nanotubes allows the virus to escape the 
      neutralizing antibody response and may contribute to the pathogenesis of viral 
      infections. The development of strategies that interfere with this process could 
      be critical in preventing the spread of viral infection.
CI  - Copyright (c) 2016, American Society for Microbiology. All Rights Reserved.
FAU - Guo, Rui
AU  - Guo R
AD  - Department of Diagnostic Medicine and Pathobiology, College of Veterinary 
      Medicine, Kansas State University, Manhattan, Kansas, USA.
FAU - Katz, Benjamin B
AU  - Katz BB
AD  - Department of Biochemistry and Molecular Biophysics, College of Arts and 
      Sciences, Kansas State University, Manhattan, Kansas, USA.
FAU - Tomich, John M
AU  - Tomich JM
AD  - Department of Biochemistry and Molecular Biophysics, College of Arts and 
      Sciences, Kansas State University, Manhattan, Kansas, USA.
FAU - Gallagher, Tom
AU  - Gallagher T
AD  - Department of Microbiology and Immunology, Stritch School of Medicine, Loyola 
      University Chicago, Maywood, Illinois, USA.
FAU - Fang, Ying
AU  - Fang Y
AD  - Department of Diagnostic Medicine and Pathobiology, College of Veterinary 
      Medicine, Kansas State University, Manhattan, Kansas, USA yfang@vet.k-state.edu.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20160429
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (Antibodies, Neutralizing)
RN  - 0 (Antibodies, Viral)
RN  - 0 (Cytoskeletal Proteins)
RN  - 0 (RNA, Viral)
RN  - 0 (Viral Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Antibodies, Neutralizing/immunology
MH  - Antibodies, Viral/immunology
MH  - Cell Line
MH  - Cytoskeletal Proteins/metabolism
MH  - Extracellular Space/physiology/*virology
MH  - Green Fluorescent Proteins
MH  - HEK293 Cells
MH  - Host-Pathogen Interactions
MH  - Humans
MH  - Intercellular Junctions/*virology
MH  - Nanotubes
MH  - Porcine respiratory and reproductive syndrome 
      virus/genetics/immunology/*physiology
MH  - RNA, Viral
MH  - Swine
MH  - Transfection
MH  - Viral Proteins/metabolism
MH  - Virion/physiology
MH  - *Virus Replication
PMC - PMC4859731
EDAT- 2016/03/18 06:00
MHDA- 2017/04/27 06:00
PMCR- 2016/10/29
CRDT- 2016/03/18 06:00
PHST- 2016/01/08 00:00 [received]
PHST- 2016/03/09 00:00 [accepted]
PHST- 2016/03/18 06:00 [entrez]
PHST- 2016/03/18 06:00 [pubmed]
PHST- 2017/04/27 06:00 [medline]
PHST- 2016/10/29 00:00 [pmc-release]
AID - JVI.00036-16 [pii]
AID - 00036-16 [pii]
AID - 10.1128/JVI.00036-16 [doi]
PST - epublish
SO  - J Virol. 2016 Apr 29;90(10):5163-5175. doi: 10.1128/JVI.00036-16. Print 2016 May 
      15.