CCR5-, DC-SIGN-dependent endocytosis and delayed reverse transcription after human immunodeficiency virus type 1 infection in human astrocytes.
We sought to determine the pathway of HIV-1 entry into human astrocytes and the fate of HIV-1 by detecting viral DNA and GFP-tagged HIV-1 in HIV-1-infected primary astrocytes. Immunochemistry and FACS analysis were used to assess the expression of DC-SIGN in human purified cultures of astrocytes. HIV-1 LTR was detected by PCR in infected cultures of human embryonic astrocytes at their third passage. GFP-Vpr-labeled R5 tropic HIV-1 was used to infect astrocytes, and was followed by confocal microscopy. Forty percent of astrocytes expressed DC-SIGN at the membrane level. Viral DNA was detected 5 days after infection in human astrocytes, but not in the presence of anti-CCR5 and anti-DC-SIGN mAbs. T20, NH4Cl, and bafilomycin had no effect on viral DNA detection. We found that 67% of the fluorescent GFP-Vpr-labeled R5 tropic HIV-1 viruses were present in the endosomes of astrocytes at 24 h, but not in the presence of anti-CCR5 or DC-SIGN mAbs. Bafilomycin and NH(4)Cl each increased the amount of fluorescent HIV-1 detected outside endosomes. Titers of p24 remained low from day 1 to day 5 postinfection, in the presence or absence of NH4Cl. Astrocytes express DC-SIGN and HIV-1 penetrates into these cells through CCR5- and/or DCSIGN- mediated endocytosis, via a pH-dependent pathway, with a delayed reverse transcription after infection without productive infection.
Deiva
Kumaran
K
Laboratoire "Immunité antivirale systémique et cérébrale," INSERM U-802, Faculté de Médecine Paris-Sud et Université Paris-Sud eleven, 94276 Le Kremlin-Bicêtre Cedex, France.
Khiati
Abdelkader
A
Hery
Christiane
C
Salim
Hassan
H
Leclerc
Philippe
P
Horellou
Philippe
P
Tardieu
Marc
M
D000911:Antibodies, Monoclonal; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D004279:DNA, Viral / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D011992:Endosomes / Q000378:metabolism; D015658:HIV Infections / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D037181:Lectins, C-Type / Q000378:metabolism*; D019713:Receptors, CCR5 / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D048348:Reverse Transcription / Q000502:physiology*; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
D000911:Antibodies, Monoclonal; D015815:Cell Adhesion Molecules; D004279:DNA, Viral; D037181:Lectins, C-Type; D019713:Receptors, CCR5; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin