Regulation of Toll-like receptor-2 expression by the Gal-lectin of Entamoeba histolytica.
The Gal/GalNAc lectin (Gal-lectin) of Entamoeba histolytica is a surface molecule involved in parasite adherence to host cells and is the most promising subunit vaccine candidate against amoebiasis. As macrophages are the major effector cells in host defense against amoebas, we studied the molecular mechanisms by which Gal-lectin activates macrophage. Microarray analysis showed that Gal-lectin up-regulated mRNAs of several cytokines and receptor genes involved in proinflammatory responses. The mechanism whereby the Gal-lectin regulates Toll-like receptor 2 (TLR-2) expression in macrophages was studied. Native Gal-lectin increased TLR-2 mRNA expression in a dose- and time-dependent fashion; peak response occurred with 1 microg/ml after 2 h stimulation. By immunoflourescence, enhanced surface expression of TLR-2 was observed after 12 h. With the use of nonoverlapping anti-Gal-lectin monoclonal antibodies that map to the carbohydrate recognition domain, amino acid 596-1082 was identified as the TLR-2 stimulating region. The Gal-lectin increased TLR-2 gene transcription, and the half-life of the mRNA transcripts was 1.4 h. Inhibition of nuclear factor (NF)-kappaB suppressed TLR-2 mRNA induction by the Gal-lectin. Moreover, cells pretreated with an inhibitor of p38 kinase (SB 208530) inhibited Gal-lectin induced TLR-2 mRNA expression by 40%. We conclude that the Gal-lectin activates NF-kappaB and MAP kinase-signaling pathways in macrophages culminating in the induction of several genes including TLR-2 and hypothesize that this could have a significant impact on macrophage activation and contribute to amoebic pathogenesis.
Kammanadiminti
Srinivas J
SJ
Institute of Parasitology of McGill University Macdonald Campus,Ste. Anne de Bellevue, Quebec, Canada.
Mann
Barbara J
BJ
Dutil
Lisa
L
Chadee
Kris
K
D000818:Animals; D002460:Cell Line; D004748:Entamoeba histolytica; D037161:Galectins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000494:pharmacology*; D005786:Gene Expression Regulation; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D008562:Membrane Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000235:genetics; D051195:Toll-Like Receptor 2; D051193:Toll-Like Receptors; D014158:Transcription, Genetic; D048051:p38 Mitogen-Activated Protein Kinases
D037161:Galectins; D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D051195:Toll-Like Receptor 2; D051193:Toll-Like Receptors; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases