Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies

Background: Dengue is the most prevalent arthropod-borne viral human disease in tropical and subtropical regions, caused by four dengue virus (DENV) serotypes. In spite of the increasing global incidence, no specific antiviral therapy is available. Cells of the mononuclear phagocyte lineage are the...

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Publicado:	2018
Materias:	acridine orange ammonium chloride antivirus agent CD32 antigen chlorpromazine cholera toxin B subunit clathrin dansylcadaverine dengue virus monoclonal antibody 2h2 dengue virus monoclonal antibody 3h5 dynamin dynasore E2 protein Fc receptor fluorescein isothiocyanate methyl beta cyclodextrin monoclonal antibody unclassified drug viral protein virus antibody virus RNA beta cyclodextrin derivative hydrazone derivative methyl-beta-cyclodextrin N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide animal cell antigen antibody complex Article bone marrow cell C6/36 cell line cell viability clathrin-coated vesicle controlled study Dengue virus 2 endocytosis endosome fluorescence microscopy human human cell infection K-562 cell line nonhuman pH quantitative analysis real time polymerase chain reaction reverse transcription polymerase chain reaction serotype U-937 cell line Vero cell line viral plaque assay virion virus entry virus infectivity virus load cell survival Dengue virus drug effect physiology virology Ammonium Chloride Antibodies, Viral beta-Cyclodextrins Cell Survival Chlorpromazine Dengue Virus Endocytosis Humans Hydrazones K562 Cells Myeloid Cells U937 Cells Virus Internalization
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19352727_v12_n8_p_Carro http://hdl.handle.net/20.500.12110/paper_19352727_v12_n8_p_Carro
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

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spelling	paper:paper_19352727_v12_n8_p_Carro2023-06-08T16:31:52Z Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies acridine orange ammonium chloride antivirus agent CD32 antigen chlorpromazine cholera toxin B subunit clathrin dansylcadaverine dengue virus monoclonal antibody 2h2 dengue virus monoclonal antibody 3h5 dynamin dynasore E2 protein Fc receptor fluorescein isothiocyanate methyl beta cyclodextrin monoclonal antibody unclassified drug viral protein virus antibody virus RNA beta cyclodextrin derivative chlorpromazine hydrazone derivative methyl-beta-cyclodextrin N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide virus antibody animal cell antigen antibody complex Article bone marrow cell C6/36 cell line cell viability clathrin-coated vesicle controlled study Dengue virus 2 endocytosis endosome fluorescence microscopy human human cell infection K-562 cell line nonhuman pH quantitative analysis real time polymerase chain reaction reverse transcription polymerase chain reaction serotype U-937 cell line Vero cell line viral plaque assay virion virus entry virus infectivity virus load bone marrow cell cell survival Dengue virus drug effect physiology virology virus entry Ammonium Chloride Antibodies, Viral beta-Cyclodextrins Cell Survival Chlorpromazine Dengue Virus Endocytosis Humans Hydrazones K562 Cells Myeloid Cells U937 Cells Virus Internalization Background: Dengue is the most prevalent arthropod-borne viral human disease in tropical and subtropical regions, caused by four dengue virus (DENV) serotypes. In spite of the increasing global incidence, no specific antiviral therapy is available. Cells of the mononuclear phagocyte lineage are the main targets either for direct antibody (Ab)-independent or Ab-mediated human DENV infection, usually associated to the severe forms of disease. Since the virus entry may be a convenient therapeutic alternative, this study aimed to investigate the mode of DENV internalization into myeloid cells in the absence and presence of DENV Ab and evaluate the inhibitory activity of diverse biochemical inhibitors of endocytosis. Methodology/principal findings: By infectivity assays and quantitative RT-PCR determinations, it was demonstrated that DENV-2 entry into U937 and K562 cells in the absence of Ab was highly inhibited by the early treatment with ammonium chloride, chlorpromazine and dynasore, but it was not affected by methyl-β-cyclodextrin, indicating that DENV-2 utilizes a low pH-dependent, clathrin- and dynamin-mediated endocytic infectious pathway for the direct entry into both human myeloid cells. To study the Ab-mediated entry of DENV, the experimental conditions for enhancement of infection were established by inoculating immune complexes formed with DENV-2 and the Ab 2H2 or 3H5. The internalization of DENV-2-2H2 or DENV-2-3H5 complexes in both myeloid cells was also dependent on acid pH and dynamin but a differential requirement of the clathrin-mediated endocytic route was observed depending on the FcγR involved in the complex uptake: the infection through FcγRII was dependent on clathrin-coated vesicles whereas the internalization pathway mediated by FcγRI was independent of clathrin. This property was not serotype-specific. Conclusions/significance: DENV entry into myeloid cells in the absence or presence of Ab can be blocked by diverse biochemical inhibitors affecting the cellular factors involved in endocytosis. The identification of the virus-host interactions involved in virus penetration may allow the finding of host-targeted antivirals widely active against diverse pathogenic flaviviruses with similar requirements for virus entry. © 2018 Carro et al. http://creativecommons.org/licenses/by/4.0/. 2018 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19352727_v12_n8_p_Carro http://hdl.handle.net/20.500.12110/paper_19352727_v12_n8_p_Carro
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	acridine orange ammonium chloride antivirus agent CD32 antigen chlorpromazine cholera toxin B subunit clathrin dansylcadaverine dengue virus monoclonal antibody 2h2 dengue virus monoclonal antibody 3h5 dynamin dynasore E2 protein Fc receptor fluorescein isothiocyanate methyl beta cyclodextrin monoclonal antibody unclassified drug viral protein virus antibody virus RNA beta cyclodextrin derivative chlorpromazine hydrazone derivative methyl-beta-cyclodextrin N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide virus antibody animal cell antigen antibody complex Article bone marrow cell C6/36 cell line cell viability clathrin-coated vesicle controlled study Dengue virus 2 endocytosis endosome fluorescence microscopy human human cell infection K-562 cell line nonhuman pH quantitative analysis real time polymerase chain reaction reverse transcription polymerase chain reaction serotype U-937 cell line Vero cell line viral plaque assay virion virus entry virus infectivity virus load bone marrow cell cell survival Dengue virus drug effect physiology virology virus entry Ammonium Chloride Antibodies, Viral beta-Cyclodextrins Cell Survival Chlorpromazine Dengue Virus Endocytosis Humans Hydrazones K562 Cells Myeloid Cells U937 Cells Virus Internalization
spellingShingle	acridine orange ammonium chloride antivirus agent CD32 antigen chlorpromazine cholera toxin B subunit clathrin dansylcadaverine dengue virus monoclonal antibody 2h2 dengue virus monoclonal antibody 3h5 dynamin dynasore E2 protein Fc receptor fluorescein isothiocyanate methyl beta cyclodextrin monoclonal antibody unclassified drug viral protein virus antibody virus RNA beta cyclodextrin derivative chlorpromazine hydrazone derivative methyl-beta-cyclodextrin N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide virus antibody animal cell antigen antibody complex Article bone marrow cell C6/36 cell line cell viability clathrin-coated vesicle controlled study Dengue virus 2 endocytosis endosome fluorescence microscopy human human cell infection K-562 cell line nonhuman pH quantitative analysis real time polymerase chain reaction reverse transcription polymerase chain reaction serotype U-937 cell line Vero cell line viral plaque assay virion virus entry virus infectivity virus load bone marrow cell cell survival Dengue virus drug effect physiology virology virus entry Ammonium Chloride Antibodies, Viral beta-Cyclodextrins Cell Survival Chlorpromazine Dengue Virus Endocytosis Humans Hydrazones K562 Cells Myeloid Cells U937 Cells Virus Internalization Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies
topic_facet	acridine orange ammonium chloride antivirus agent CD32 antigen chlorpromazine cholera toxin B subunit clathrin dansylcadaverine dengue virus monoclonal antibody 2h2 dengue virus monoclonal antibody 3h5 dynamin dynasore E2 protein Fc receptor fluorescein isothiocyanate methyl beta cyclodextrin monoclonal antibody unclassified drug viral protein virus antibody virus RNA beta cyclodextrin derivative chlorpromazine hydrazone derivative methyl-beta-cyclodextrin N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide virus antibody animal cell antigen antibody complex Article bone marrow cell C6/36 cell line cell viability clathrin-coated vesicle controlled study Dengue virus 2 endocytosis endosome fluorescence microscopy human human cell infection K-562 cell line nonhuman pH quantitative analysis real time polymerase chain reaction reverse transcription polymerase chain reaction serotype U-937 cell line Vero cell line viral plaque assay virion virus entry virus infectivity virus load bone marrow cell cell survival Dengue virus drug effect physiology virology virus entry Ammonium Chloride Antibodies, Viral beta-Cyclodextrins Cell Survival Chlorpromazine Dengue Virus Endocytosis Humans Hydrazones K562 Cells Myeloid Cells U937 Cells Virus Internalization
description	Background: Dengue is the most prevalent arthropod-borne viral human disease in tropical and subtropical regions, caused by four dengue virus (DENV) serotypes. In spite of the increasing global incidence, no specific antiviral therapy is available. Cells of the mononuclear phagocyte lineage are the main targets either for direct antibody (Ab)-independent or Ab-mediated human DENV infection, usually associated to the severe forms of disease. Since the virus entry may be a convenient therapeutic alternative, this study aimed to investigate the mode of DENV internalization into myeloid cells in the absence and presence of DENV Ab and evaluate the inhibitory activity of diverse biochemical inhibitors of endocytosis. Methodology/principal findings: By infectivity assays and quantitative RT-PCR determinations, it was demonstrated that DENV-2 entry into U937 and K562 cells in the absence of Ab was highly inhibited by the early treatment with ammonium chloride, chlorpromazine and dynasore, but it was not affected by methyl-β-cyclodextrin, indicating that DENV-2 utilizes a low pH-dependent, clathrin- and dynamin-mediated endocytic infectious pathway for the direct entry into both human myeloid cells. To study the Ab-mediated entry of DENV, the experimental conditions for enhancement of infection were established by inoculating immune complexes formed with DENV-2 and the Ab 2H2 or 3H5. The internalization of DENV-2-2H2 or DENV-2-3H5 complexes in both myeloid cells was also dependent on acid pH and dynamin but a differential requirement of the clathrin-mediated endocytic route was observed depending on the FcγR involved in the complex uptake: the infection through FcγRII was dependent on clathrin-coated vesicles whereas the internalization pathway mediated by FcγRI was independent of clathrin. This property was not serotype-specific. Conclusions/significance: DENV entry into myeloid cells in the absence or presence of Ab can be blocked by diverse biochemical inhibitors affecting the cellular factors involved in endocytosis. The identification of the virus-host interactions involved in virus penetration may allow the finding of host-targeted antivirals widely active against diverse pathogenic flaviviruses with similar requirements for virus entry. © 2018 Carro et al. http://creativecommons.org/licenses/by/4.0/.
title	Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies
title_short	Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies
title_full	Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies
title_fullStr	Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies
title_full_unstemmed	Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies
title_sort	blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies
publishDate	2018
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19352727_v12_n8_p_Carro http://hdl.handle.net/20.500.12110/paper_19352727_v12_n8_p_Carro
_version_	1768546650619379712

Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies

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