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Abstract:

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/.

Registro:

Documento: Artículo
Título:Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies
Autor:Carro, A.C.; Piccini, L.E.; Damonte, E.B.
Filiación:Laboratory of Virology, Department of Biological Chemistry, Faculty of Sciences, University of Buenos Aires, Buenos Aires, Argentina
IQUIBICEN, National Research Council (CONICET, Department of Biological Chemistry, University of Buenos Aires, Ciudad Universitaria, Buenos Aires, Argentina
Technology Transfer Office, CCT Patagonia Norte, National Research Council, San Carlos de Bariloche, Argentina
Palabras clave: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
Año:2018
Volumen:12
Número:8
DOI: http://dx.doi.org/10.1371/journal.pntd.0006685
Título revista:PLoS Neglected Tropical Diseases
Título revista abreviado:PLoS. Negl. Trop. Dis.
ISSN:19352727
CAS:acridine orange, 494-38-2, 65-61-2; ammonium chloride, 12125-02-9; chlorpromazine, 50-53-3, 69-09-0; dansylcadaverine, 10121-91-2; dynamin; dynasore, 1202867-00-2; fluorescein isothiocyanate, 25168-13-2, 27072-45-3, 3326-32-7; Ammonium Chloride; Antibodies, Viral; beta-Cyclodextrins; Chlorpromazine; Hydrazones; methyl-beta-cyclodextrin; N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19352727_v12_n8_p_Carro

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Citas:

---------- APA ----------
Carro, A.C., Piccini, L.E. & Damonte, E.B. (2018) . Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies. PLoS Neglected Tropical Diseases, 12(8).
http://dx.doi.org/10.1371/journal.pntd.0006685
---------- CHICAGO ----------
Carro, A.C., Piccini, L.E., Damonte, E.B. "Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies" . PLoS Neglected Tropical Diseases 12, no. 8 (2018).
http://dx.doi.org/10.1371/journal.pntd.0006685
---------- MLA ----------
Carro, A.C., Piccini, L.E., Damonte, E.B. "Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies" . PLoS Neglected Tropical Diseases, vol. 12, no. 8, 2018.
http://dx.doi.org/10.1371/journal.pntd.0006685
---------- VANCOUVER ----------
Carro, A.C., Piccini, L.E., Damonte, E.B. Blockade of dengue virus entry into myeloid cells by endocytic inhibitors in the presence or absence of antibodies. PLoS. Negl. Trop. Dis. 2018;12(8).
http://dx.doi.org/10.1371/journal.pntd.0006685