Invasion of HeLa cells by Enterococcus faecalis clinical isolates.
We examined the in vitro ability of Enterococcus faecalis clinical isolates to adhere to and to invade HeLa cells, suggested to be a valuable model system to study bacteria-directed endocytosis. Using a variety of compounds that act on eukaryotic cell structures, both microtubules and microfilaments were found to be involved in enterococcal entry into cells. Two distinct modes of interaction were observed: in one, a close proximity of bacteria with the cell membrane was observed, possibly leading to direct engulfment of the bacterial cell. In the other mode, cellular pseudopodal formation seemed to be stimulated by vicinity of bacterial cells; in some cases, such associations involved formation of clathrin-coated-like vesicles before internalizing enterococci. The above-mentioned experimental data together with the use of monodansylcadaverine, amiloride and NH4Cl, all involved in cytosol acidification and inhibition of receptor-mediated endocytosis (RME), led us to conclude that E. faecalis is internalized within HeLa cells by more than one invasion pathway. One, sensitive to amiloride, is most likely a macropinocytic, actin-dependent uptake mechanism, which determines the production of large smooth-membrane vacuoles engulfing enterococci. The other is RME, in which entry is dependent on both microfilament and microtubule structural integrity.
Laboratorio di Ultrastrutture, Istituto Superiore di Sanità, Rome, Italy. baldassa@iss.it
Lucia
L
Bertuccini
Maria Grazia
MG
Ammendolia
Fabiana
F
Superti
Lucilla
L
Baldassarri
D000143:Acids / Q000037:antagonists & inhibitors / Q000737:chemistry; D001422:Bacterial Adhesion / Q000502:physiology; D002103:Cadaverine / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D002478:Cells, Cultured; D003078:Colchicine / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D015638:Cytochalasin D / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D004705:Endocytosis; D013293:Enterococcus faecalis / Q000302:isolation & purification / Q000378:metabolism / Q000472:pathogenicity*; D016908:Gram-Positive Bacterial Infections / Q000382:microbiology; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron
D000143:Acids; D015638:Cytochalasin D; C008542:monodansylcadaverine; D002103:Cadaverine; D003078:Colchicine