The interplay between Entamoeba and enteropathogenic bacteria modulates epithelial cell damage.
BACKGROUND
Mixed intestinal infections with Entamoeba histolytica, Entamoeba dispar and bacteria with exacerbated manifestations of disease are common in regions where amoebiasis is endemic. However, amoeba-bacteria interactions remain largely unexamined.


METHODS
Trophozoites of E. histolytica and E. dispar were co-cultured with enteropathogenic bacteria strains Escherichia coli (EPEC), Shigella dysenteriae and a commensal Escherichia coli. Amoebae that phagocytosed bacteria were tested for a cytopathic effect on epithelial cell monolayers. Cysteine proteinase activity, adhesion and cell surface concentration of Gal/GalNAc lectin were analyzed in amoebae showing increased virulence. Structural and functional changes and induction of IL-8 expression were determined in epithelial cells before and after exposure to bacteria. Chemotaxis of amoebae and neutrophils to human IL-8 and conditioned culture media from epithelial cells exposed to bacteria was quantified.


RESULTS
E. histolytica digested phagocytosed bacteria, although S. dysenteriae retained 70% viability after ingestion. Phagocytosis of pathogenic bacteria augmented the cytopathic effect of E. histolytica and increased expression of Gal/GalNAc lectin on the amoebic surface and increased cysteine proteinase activity. E. dispar remained avirulent. Adhesion of amoebae and damage to cells exposed to bacteria were increased. Additional increases were observed if amoebae had phagocytosed bacteria. Co-culture of epithelial cells with enteropathogenic bacteria disrupted monolayer permeability and induced expression of IL-8. Media from these co-cultures and human recombinant IL-8 were similarly chemotactic for neutrophils and E. histolytica.


CONCLUSIONS
Epithelial monolayers exposed to enteropathogenic bacteria become more susceptible to E. histolytica damage. At the same time, phagocytosis of pathogenic bacteria by amoebae further increased epithelial cell damage.


CONCLUSIONS
The in vitro system presented here provides evidence that the Entamoeba/enteropathogenic bacteria interplay modulates epithelial cell responses to the pathogens. In mixed intestinal infections, where such interactions are possible, they could influence the outcome of disease. The results offer insights to continue research on this phenomenon.
Galván-Moroyoqui
José Manuel
JM
Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, México DF, México.
Del Carmen Domínguez-Robles
M
M
Franco
Elizabeth
E
Meza
Isaura
I
D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D002633:Chemotaxis; D057056:Cysteine Proteases / Q000235:genetics / Q000378:metabolism; D004285:Dogs; D004405:Dysentery, Bacillary / Q000276:immunology / Q000382:microbiology; D004747:Entamoeba / Q000201:enzymology / Q000382:microbiology / Q000472:pathogenicity / Q000502:physiology*; D004749:Entamoebiasis / Q000276:immunology / Q000469:parasitology; D054308:Enteropathogenic Escherichia coli / Q000472:pathogenicity / Q000502:physiology*; D004847:Epithelial Cells / Q000276:immunology / Q000382:microbiology* / Q000469:parasitology*; D004927:Escherichia coli Infections / Q000276:immunology / Q000382:microbiology; D006801:Humans; D010587:Phagocytosis; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism; D012762:Shigella dysenteriae / Q000472:pathogenicity / Q000502:physiology*; D014774:Virulence
D015800:Protozoan Proteins; D057056:Cysteine Proteases