Mechanism of enhancement of the antiviral action of interferon against herpes simplex virus-1 by chloroquine.
Using double immunofluorescence, we have shown previously that interferon (IFN) treatment inhibits the transport of herpes simplex virus-1 (HSV-1) gD from the Golgi complex to the plasma membrane in the virus infected and gD cDNA transfected LMtk-cells. In the present study, we quantitated the gD protein on the cell surface and localized the gD protein in the trans-Golgi network (TGN). The results showed 10-fold less fluorescence for the gD protein on the cell surface in IFN-treated LMtk-cells. Subcellular fractionation studies demonstrated that gD was associated with TGN-enriched membranes. Gold labeling for DAMP distribution using electron microscopy showed that IFN raised the pH of TGN. IFNs induced alkalinization of TGN may be related to the block in the transport of HSV-1 gD. Earlier we reported that a subeffective dose of chloroquine (CHL) or IFN does not change the pHi. However, both CHL and IFN together raise the pHi significantly. To study the biologic significance of the finding, the effect of these subeffective doses of IFN and CHL on the antiviral activity and the transport of the gD protein was studied. Results suggested that CHL enhance the antiviral activity of IFN against HSV-1 and concomitantly increase the inhibition of HSV-1 gD transport. This IFN-induced increase in pHi of the TGN may also explain the inhibitory effect of IFN reported on the terminal steps of some of the enveloped viruses.
Singh
A K
AK
Uniformed Services University of the Health Sciences, Bethesda, MD, USA.
Sidhu
G S
GS
Friedman
R M
RM
Maheshwari
R K
RK
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology*; D004357:Drug Synergism; D005434:Flow Cytometry; D018259:Herpesvirus 1, Human / Q000187:drug effects*; D007150:Immunohistochemistry; D016899:Interferon-beta / Q000494:pharmacology*; D051379:Mice; D008854:Microscopy, Electron; D011994:Recombinant Proteins / Q000494:pharmacology; D012016:Reference Values; D013347:Subcellular Fractions / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000096:biosynthesis*
D000998:Antiviral Agents; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C044768:glycoprotein E, herpes simplex virus type 1; D016899:Interferon-beta; D002738:Chloroquine