Human immunodeficiency virus type 1 enters brain microvascular endothelia by macropinocytosis dependent on lipid rafts and the mitogen-activated protein kinase signaling pathway

Nancy Q. Liu, Albert S. Lossinsky, Waldemar Popik, Xia Li, Chandrasekhar Gujuluva, Benjamin Kriederman, Jaclyn Roberts, Tatania Pushkarsky, Michael Bukrinsky, Marlys H Witte, Martin E Weinand, Milan Fiala

Research output: Contribution to journalArticle

223 Citations (Scopus)

Abstract

Brain microvascular endothelial cells (BMVECs) present an incomplete barrier to human immunodeficiency virus type 1 (HIV-1) neuroinvasion. In order to clarify the mechanisms of HIV-1 invasion, we have examined HIV-1 uptake and transcellular penetration in an in vitro BMVEC model. No evidence of productive infection was observed by luciferase, PCR, and reverse transcriptase assays. Approximately 1% of viral RNA and 1% of infectious virus penetrated the BMVEC barrier without disruption of tight junctions. The virus upregulated ICAM-1 on plasma membranes and in cytoplasmic vesiculotubular structures. HIV-1 virions were entangled by microvilli and were taken into cytoplasmic vesicles through surface invaginations without fusion of the virus envelope with the plasma membrane. Subsequently, the cytoplasmic vesicles fused with lysosomes, the virions were lysed, and the vesicles diminished in size. Upon cell entry, HIV-1 colocalized with cholera toxin B, which targets lipid raft-associated GM1 ganglioside. Cholesterol-extracting agents, cyclodextrin and nystatin, and polyanion heparin significantly inhibited virus entry. Anti-CD4 had no effect and the chemokine AOP-RANTES had only a slight inhibitory effect on virus entry. HIV-1 activated the mitogen-activated protein kinase (MAPK) pathway, and inhibition of MAPK/Erk kinase inhibited virus entry. Entry was also blocked by dimethylamiloride, indicating that HIV-1 enters endothelial cells by macropinocytosis. Therefore, HIV-1 penetrates BMVECs in ICAM-1-lined macropinosomes by a mechanism involving lipid rafts, MAPK signaling, and glycosylaminoglycans, while CD4 and chemokine receptors play limited roles in this process.

Original languageEnglish (US)
Pages (from-to)6689-6700
Number of pages12
JournalJournal of Virology
Volume76
Issue number13
DOIs
StatePublished - 2002
Externally publishedYes

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Human immunodeficiency virus 1
Mitogen-Activated Protein Kinases
endothelium
mitogen-activated protein kinase
Endothelium
HIV-1
brain
Lipids
endothelial cells
Brain
lipids
Endothelial Cells
Virus Internalization
viruses
Cytoplasmic Vesicles
Intercellular Adhesion Molecule-1
Viruses
virion
Virion
plasma membrane

ASJC Scopus subject areas

  • Immunology

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Human immunodeficiency virus type 1 enters brain microvascular endothelia by macropinocytosis dependent on lipid rafts and the mitogen-activated protein kinase signaling pathway. / Liu, Nancy Q.; Lossinsky, Albert S.; Popik, Waldemar; Li, Xia; Gujuluva, Chandrasekhar; Kriederman, Benjamin; Roberts, Jaclyn; Pushkarsky, Tatania; Bukrinsky, Michael; Witte, Marlys H; Weinand, Martin E; Fiala, Milan.

In: Journal of Virology, Vol. 76, No. 13, 2002, p. 6689-6700.

Research output: Contribution to journalArticle

Liu, Nancy Q. ; Lossinsky, Albert S. ; Popik, Waldemar ; Li, Xia ; Gujuluva, Chandrasekhar ; Kriederman, Benjamin ; Roberts, Jaclyn ; Pushkarsky, Tatania ; Bukrinsky, Michael ; Witte, Marlys H ; Weinand, Martin E ; Fiala, Milan. / Human immunodeficiency virus type 1 enters brain microvascular endothelia by macropinocytosis dependent on lipid rafts and the mitogen-activated protein kinase signaling pathway. In: Journal of Virology. 2002 ; Vol. 76, No. 13. pp. 6689-6700.
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abstract = "Brain microvascular endothelial cells (BMVECs) present an incomplete barrier to human immunodeficiency virus type 1 (HIV-1) neuroinvasion. In order to clarify the mechanisms of HIV-1 invasion, we have examined HIV-1 uptake and transcellular penetration in an in vitro BMVEC model. No evidence of productive infection was observed by luciferase, PCR, and reverse transcriptase assays. Approximately 1{\%} of viral RNA and 1{\%} of infectious virus penetrated the BMVEC barrier without disruption of tight junctions. The virus upregulated ICAM-1 on plasma membranes and in cytoplasmic vesiculotubular structures. HIV-1 virions were entangled by microvilli and were taken into cytoplasmic vesicles through surface invaginations without fusion of the virus envelope with the plasma membrane. Subsequently, the cytoplasmic vesicles fused with lysosomes, the virions were lysed, and the vesicles diminished in size. Upon cell entry, HIV-1 colocalized with cholera toxin B, which targets lipid raft-associated GM1 ganglioside. Cholesterol-extracting agents, cyclodextrin and nystatin, and polyanion heparin significantly inhibited virus entry. Anti-CD4 had no effect and the chemokine AOP-RANTES had only a slight inhibitory effect on virus entry. HIV-1 activated the mitogen-activated protein kinase (MAPK) pathway, and inhibition of MAPK/Erk kinase inhibited virus entry. Entry was also blocked by dimethylamiloride, indicating that HIV-1 enters endothelial cells by macropinocytosis. Therefore, HIV-1 penetrates BMVECs in ICAM-1-lined macropinosomes by a mechanism involving lipid rafts, MAPK signaling, and glycosylaminoglycans, while CD4 and chemokine receptors play limited roles in this process.",
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AU - Liu, Nancy Q.

AU - Lossinsky, Albert S.

AU - Popik, Waldemar

AU - Li, Xia

AU - Gujuluva, Chandrasekhar

AU - Kriederman, Benjamin

AU - Roberts, Jaclyn

AU - Pushkarsky, Tatania

AU - Bukrinsky, Michael

AU - Witte, Marlys H

AU - Weinand, Martin E

AU - Fiala, Milan

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AB - Brain microvascular endothelial cells (BMVECs) present an incomplete barrier to human immunodeficiency virus type 1 (HIV-1) neuroinvasion. In order to clarify the mechanisms of HIV-1 invasion, we have examined HIV-1 uptake and transcellular penetration in an in vitro BMVEC model. No evidence of productive infection was observed by luciferase, PCR, and reverse transcriptase assays. Approximately 1% of viral RNA and 1% of infectious virus penetrated the BMVEC barrier without disruption of tight junctions. The virus upregulated ICAM-1 on plasma membranes and in cytoplasmic vesiculotubular structures. HIV-1 virions were entangled by microvilli and were taken into cytoplasmic vesicles through surface invaginations without fusion of the virus envelope with the plasma membrane. Subsequently, the cytoplasmic vesicles fused with lysosomes, the virions were lysed, and the vesicles diminished in size. Upon cell entry, HIV-1 colocalized with cholera toxin B, which targets lipid raft-associated GM1 ganglioside. Cholesterol-extracting agents, cyclodextrin and nystatin, and polyanion heparin significantly inhibited virus entry. Anti-CD4 had no effect and the chemokine AOP-RANTES had only a slight inhibitory effect on virus entry. HIV-1 activated the mitogen-activated protein kinase (MAPK) pathway, and inhibition of MAPK/Erk kinase inhibited virus entry. Entry was also blocked by dimethylamiloride, indicating that HIV-1 enters endothelial cells by macropinocytosis. Therefore, HIV-1 penetrates BMVECs in ICAM-1-lined macropinosomes by a mechanism involving lipid rafts, MAPK signaling, and glycosylaminoglycans, while CD4 and chemokine receptors play limited roles in this process.

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