RESUMEN
Targeting nanoparticles to desired intracellular compartments is a major challenge. Integrin-type adhesion receptors are connected to different endocytosis routes in a receptor-specific manner. According to our previous observations, the internalization of an α2ß1-integrin-echovirus-1 complex takes place via a macropinocytosis-like mechanism, suggesting that the receptor could be used to target nanoparticles to this specific entry route. Here, silica-based nanoparticles, carrying monoclonal antibodies against the α2ß1 integrin as address labels, were synthesized. Studies with flow cytometry, atomic force microscopy and confocal microscopy showed the particles to attach to the cell surface via the α2ß1 integrin. Furthermore, quantitative analysis of nanoparticle trafficking inside the cell performed with the BioImageXD software indicated that the particles enter cells via a macropinocytosis-like process and end up in caveolin-1 positive structures. Thus, we suggest that different integrins can guide particles to distinct endocytosis routes and, subsequently, also to specific intracellular compartments. In addition, we show that with the BioImageXD software it is possible to conduct sensitive and complex analyses of the behavior of small fluorescent particles inside cells, using basic confocal microscopy images.
Asunto(s)
Integrina alfa2beta1/química , Nanopartículas/química , Anticuerpos Inmovilizados/química , Anticuerpos Inmovilizados/metabolismo , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales/inmunología , Caveolina 1/metabolismo , Línea Celular Tumoral , Endocitosis , Enterovirus Humano B/genética , Enterovirus Humano B/metabolismo , Humanos , Inmunohistoquímica , Integrina alfa2beta1/inmunología , Integrina alfa2beta1/metabolismo , Microscopía de Fuerza Atómica , Microscopía Confocal , Dióxido de Silicio/químicaRESUMEN
This study was conducted to examine the ability of selected dairy strains of lactic acid bacteria to remove aflatoxin B1 (AFB1) from liquid media. Both Lactobacillus rhamnosus strain GG (LBGG) and L. rhamnosus strain LC-705 (LC705) can significantly (P > 0.05) remove AFB1 when compared with that by other strains of either Gram-positive or Gram-negative bacteria. Removal of AFB1 by LBGG and LC705 was a rapid process with approximately 80% AFB1 removed at 0 hr. Removal of AFB1 by these two strains was both temperature and bacterial concentration dependent.