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COMPARISON OF DENGUE INFECTION IN HUMAN MONONUCLEAR LEUKOCYTES WITH MOSQUITO C6/36 AND MAMMALIAN VERO CELLS USING FLOW CYTOMETRY TO DETECT VIRUS ANTIGEN
Citometria de fluxo
Células vero
Leucócitos mononucleares
C6/36
Author
Affilliation
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Protozoologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Virologia. Rio de Janeiro, RJ, Brasil.
Abstract
Fluorescent activated cell sorter (FACS) analysis is useful for the detection of cellular surface
antigens and intracellular proteins. We used this methodology in order to detect and quantify dengue
antigens in highly susceptible cells such as clone C6/36 (Aedes albopictus) and Vero cells (green
monkey kidney). Additionally, we analyzed the infection in vitro of human peripheral blood mononuclear leukocytes (PBML).
FACS analysis turned out to be a reliable technique to quantify virus growth in traditional cell
cultures of C6/36 as well as Vero cells. High rates of infection were achieved with a good statistical
correlation between the virus amount used in infection and the percentage of dengue antigen containing cells detected in infected cultures.
We also showed that human monocytes (CD14+) are preferred target cells for in vitro dengue
infection among PBML. Monocytes were much less susceptible to virus infection than cell lines but they
displayed dengue antigens detected by FACS five days after infection. In contrast, lymphocytes showed
no differences in their profile for dengue specific immunofluorescence.
Without an animal model to reproduce dengue disease, alternative assays have been sought to
correlate viral virulence with clinical manifestations and disease severity. Study of in vitro interaction
of virus and host cells may highlight this relationship.
Keywords in Portuguese
Vírus da DengueCitometria de fluxo
Células vero
Leucócitos mononucleares
C6/36
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