PMID- 20421949
OWN - NLM
STAT- MEDLINE
DCOM- 20100625
LR  - 20211020
IS  - 1553-7374 (Electronic)
IS  - 1553-7366 (Print)
IS  - 1553-7366 (Linking)
VI  - 6
IP  - 4
DP  - 2010 Apr 22
TI  - A differential role for macropinocytosis in mediating entry of the two forms of 
      vaccinia virus into dendritic cells.
PG  - e1000866
LID - 10.1371/journal.ppat.1000866 [doi]
LID - e1000866
AB  - Vaccinia virus (VACV) is being developed as a recombinant viral vaccine vector 
      for several key pathogens. Dendritic cells (DCs) are specialised antigen 
      presenting cells that are crucial for the initiation of primary immune responses; 
      however, the mechanisms of uptake of VACV by these cells are unclear. Therefore 
      we examined the binding and entry of both the intracellular mature virus (MV) and 
      extracellular enveloped virus (EV) forms of VACV into vesicular compartments of 
      monocyte-derived DCs. Using a panel of inhibitors, flow cytometry and confocal 
      microscopy we have shown that neither MV nor EV binds to the highly expressed 
      C-type lectin receptors on DCs that are responsible for capturing many other 
      viruses. We also found that both forms of VACV enter DCs via a clathrin-, 
      caveolin-, flotillin- and dynamin-independent pathway that is dependent on actin, 
      intracellular calcium and host-cell cholesterol. Both MV and EV entry were 
      inhibited by the macropinocytosis inhibitors rottlerin and dimethyl amiloride and 
      depended on phosphotidylinositol-3-kinase (PI(3)K), and both colocalised with 
      dextran but not transferrin. VACV was not delivered to the classical 
      endolysosomal pathway, failing to colocalise with EEA1 or Lamp2. Finally, 
      expression of early viral genes was not affected by bafilomycin A, indicating 
      that the virus does not depend on low pH to deliver cores to the cytoplasm. From 
      these collective results we conclude that VACV enters DCs via macropinocytosis. 
      However, MV was consistently less sensitive to inhibition and is likely to 
      utilise at least one other entry pathway. Definition and future manipulation of 
      these pathways may assist in enhancing the activity of recombinant vaccinia 
      vectors through effects on antigen presentation.
FAU - Sandgren, Kerrie J
AU  - Sandgren KJ
AD  - Centre for Virus Research, Westmead Millennium Institute, Sydney, New South 
      Wales, Australia.
FAU - Wilkinson, John
AU  - Wilkinson J
FAU - Miranda-Saksena, Monica
AU  - Miranda-Saksena M
FAU - McInerney, Gerald M
AU  - McInerney GM
FAU - Byth-Wilson, Karen
AU  - Byth-Wilson K
FAU - Robinson, Phillip J
AU  - Robinson PJ
FAU - Cunningham, Anthony L
AU  - Cunningham AL
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100422
PL  - United States
TA  - PLoS Pathog
JT  - PLoS pathogens
JID - 101238921
SB  - IM
MH  - Blotting, Western
MH  - Cell Separation
MH  - Dendritic Cells/*metabolism/*virology
MH  - Flow Cytometry
MH  - Humans
MH  - Microscopy, Confocal
MH  - Pinocytosis/*physiology
MH  - Polymerase Chain Reaction
MH  - Vaccinia virus/*metabolism
MH  - Virus Attachment
MH  - *Virus Internalization
PMC - PMC2858709
COIS- The authors have declared that no competing interests exist.
EDAT- 2010/04/28 06:00
MHDA- 2010/06/26 06:00
PMCR- 2010/04/22
CRDT- 2010/04/28 06:00
PHST- 2009/09/08 00:00 [received]
PHST- 2010/03/22 00:00 [accepted]
PHST- 2010/04/28 06:00 [entrez]
PHST- 2010/04/28 06:00 [pubmed]
PHST- 2010/06/26 06:00 [medline]
PHST- 2010/04/22 00:00 [pmc-release]
AID - 09-PLPA-RA-1584R3 [pii]
AID - 10.1371/journal.ppat.1000866 [doi]
PST - epublish
SO  - PLoS Pathog. 2010 Apr 22;6(4):e1000866. doi: 10.1371/journal.ppat.1000866.