title: Participation of the serine-rich Entamoeba histolytica protein in amebic phagocytosis of apoptotic host cells.
abstract: Entamoeba histolytica is an intestinal ameba that causes dysentery and liver abscesses. Cytotoxicity and phagocytosis of host cells characterize invasive E. histolytica infection. Prior to phagocytosis of host cells, E. histolytica induces apoptotic host cell death, using a mechanism that requires contact via an amebic galactose-specific lectin. However, lectin inhibition only partially blocks phagocytosis of already dead cells, implicating at least one additional receptor in phagocytosis. To identify receptors for engulfment of apoptotic cells, monoclonal antibodies against E. histolytica membrane antigens were screened for inhibition of phagocytosis. Of 43 antibodies screened, one blocked lectin-independent uptake of apoptotic cells, with >90% inhibition at a dose of 20 microg/ml (P < 0.0003 versus control). The same antibody also inhibited adherence to apoptotic lymphocytes and, to a lesser extent, adherence to and killing of viable lymphocytes. The antigen recognized by the inhibitory antibody was purified by affinity chromatography and identified by liquid chromatography-mass spectrometry as the serine-rich E. histolytica protein (SREHP). Consistent with this, the inhibitory antibody bound to recombinant SREHP present in bacterial lysates on immunoblots. The SREHP is an abundant immunogenic surface protein of unclear function. The results of this unbiased antibody screen strongly implicate the SREHP as a participant in E. histolytica phagocytosis and suggest that it may play an important role in adherence to apoptotic cells.
authors: Department of Medicine, University of Vermont College of Medicine, Burlington, VT 05405, USA.
authors: Jose E
authors: JE
authors: Teixeira
authors: Christopher D
authors: CD
authors: Huston
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D000913:Antibodies, Protozoan / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D002846:Chromatography, Affinity; D004748:Entamoeba histolytica / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D018613:Microscopy, Confocal; D010587:Phagocytosis; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*
chemical_list: D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D008565:Membrane Proteins; D015800:Protozoan Proteins; C073020:serine-rich protein, Entamoeba histolytica