Comparison of dengue infection in human mononuclear leukocytes with mosquito C6/36 and mammalian Vero cells using flow cytometry to detect virus antigen.
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.
Sydow
F F
FF
Departamento de Protozoologia, Instituto Oswaldo Cruz, Rio de Janeiro, Brasil.
Santiago
M A
MA
Neves-Souza
P C
PC
Cerqueira
D I
DI
Gouvea
A S
AS
Lavatori
M F
MF
Bertho
A L
AL
Kubelka
C F
CF
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis*; D002460:Cell Line / Q000821:virology; D002469:Cell Separation; D002522:Chlorocebus aethiops; D002999:Clone Cells / Q000276:immunology; D003715:Dengue / Q000276:immunology*; D003716:Dengue Virus / Q000254:growth & development / Q000276:immunology* / Q000302:isolation & purification; D005434:Flow Cytometry; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000276:immunology* / Q000821:virology; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D014709:Vero Cells / Q000166:cytology / Q000821:virology
D000956:Antigens, Viral; D018950:Lipopolysaccharide Receptors