Biochemical and cellular mechanisms regulating Acanthamoeba castellanii adherence to host cells.
Free-living amoebae belonging to the genus Acanthamoeba are the causative agents of infections such as amoebic keratitis (AK), granulomatous amoebic encephalitis (GAE) and cutaneous lesions. The mechanisms involved in the establishment of infection are unknown. However, it is accepted that the initial phase of pathogenesis involves adherence to the host tissue. In this work, we analysed surface molecules with an affinity for epithelial and neuronal cells from the trophozoites of Acanthamoeba castellanii. We also investigated the cellular mechanisms that govern the process of trophozoite adhesion to the host cells. We first used confocal and epifluorescence microscopy to examine the distribution of the A. castellanii actin cytoskeleton during interaction with the host cells. The use of drugs, as cytochalasin B (CB) and latrunculin B (LB), revealed the participation of cytoskeletal filaments in the adhesion process. In addition, to identify the proteins and glycoproteins on the surface of A. castellanii, the trophozoites were labelled with biotin and biotinylated lectins. The results revealed bands of surface proteins, some of which were glycoproteins with mannose and N-acetylglucosamine residues. Interaction assays of biotinylated amoebae proteins with epithelial and neuronal cells showed that some surface proteins had affinity for both cell types. The results of this study provide insight into the biochemical and cellular mechanisms of the Acanthamoeba infection process.
Departamento de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta S/N Col. Noria Alta, C.P. 36050, Guanajuato, Guanajuato, México.
K J
KJ
Soto-Arredondo
Departamento de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta S/N Col. Noria Alta, C.P. 36050, Guanajuato, Guanajuato, México.
L L
LL
Flores-Villavicencio
Departamento de Biología Celular, Centro de Investigación y Estudios Avanzados, Instituto Politécnico Nacional, Av. Instituto Politecnico Nacional 2508, San Pedro Zacatenco, Gustavo A. Madero, 07360, Ciudad de México, México.
J J
JJ
Serrano-Luna
Departamento de Infectómica y Patogénesis Molecular, Centro de Investigación y Estudios Avanzados, Instituto Politécnico Nacional, Av. Instituto Politécnico Nacional 2508, San Pedro Zacatenco, Gustavo A. Madero, 07360, Ciudad de México, México.
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M
Shibayama
Departamento de Biología, División de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta S/N Col. Noria Alta, C.P. 36050, Guanajuato, Guanajuato, México.
M
M
Sabanero-López
D048248:Acanthamoeba castellanii / Q000187:drug effects / Q000472:pathogenicity / Q000502:physiology*; D000117:Acetylglucosamine / Q000378:metabolism; D008841:Actin Cytoskeleton / Q000378:metabolism; D000562:Amebiasis / Q000469:parasitology*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002448:Cell Adhesion; D002460:Cell Line; D022081:Cell Surface Extensions / Q000378:metabolism; D003571:Cytochalasin B / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism*; D004847:Epithelial Cells / Q000469:parasitology; D006023:Glycoproteins / Q000378:metabolism; D006790:Host-Parasite Interactions; D006801:Humans; D037102:Lectins / Q000378:metabolism; D008358:Mannose / Q000378:metabolism; D008954:Models, Biological; D015800:Protozoan Proteins / Q000378:metabolism; D053778:Thiazolidines / Q000494:pharmacology; D053058:Trophozoites
D019086:Bridged Bicyclo Compounds, Heterocyclic; D006023:Glycoproteins; D037102:Lectins; D015800:Protozoan Proteins; D053778:Thiazolidines; D003571:Cytochalasin B; C037068:latrunculin B; D008358:Mannose; D000117:Acetylglucosamine