Fluorescent membrane markers elucidate the association of Borrelia burgdorferi with tick cell lines
Braz. j. med. biol. res
; 49(7): e5211, 2016. tab, graf
Article
en En
| LILACS
| ID: lil-785057
Biblioteca responsable:
BR1.1
ABSTRACT
This study aimed to describe the association of Borrelia burgdorferi s.s. with ixodid tick cell lines by flow cytometry and fluorescence and confocal microscopy. Spirochetes were stained with a fluorescent membrane marker (PKH67 or PKH26), inoculated into 8 different tick cell lines and incubated at 30°C for 24 h. PKH efficiently stained B. burgdorferi without affecting bacterial viability or motility. Among the tick cell lines tested, the Rhipicephalus appendiculatus cell line RA243 achieved the highest percentage of association/internalization, with both high (90%) and low (10%) concentrations of BSK-H medium in tick cell culture medium. Treatment with cytochalasin D dramatically reduced the average percentage of cells with internalized spirochetes, which passed through a dramatic morphological change during their internalization by the host cell as observed in time-lapse photography. Almost all of the fluorescent bacteria were seen to be inside the tick cells. PKH labeling of borreliae proved to be a reliable and valuable tool to analyze the association of spirochetes with host cells by flow cytometry, confocal and fluorescence microscopy.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
LILACS
Asunto principal:
Coloración y Etiquetado
/
Garrapatas
/
Borrelia burgdorferi
Tipo de estudio:
Risk_factors_studies
Límite:
Animals
Idioma:
En
Revista:
Braz. j. med. biol. res
Asunto de la revista:
BIOLOGIA
/
MEDICINA
Año:
2016
Tipo del documento:
Article
País de afiliación:
Brasil
Pais de publicación:
Brasil