Role of beta-D-galactofuranose in Leishmania major macrophage invasion.
The role of glycosylinositol phospholipid 1 (GIPL-1) of Leishmania (Leishmania) major in the interaction of promastigotes and amastigotes with macrophages was analyzed. Monoclonal antibody MEST-1, which recognizes glycolipids containing terminal galactofuranose (Galf) residues (E. Suzuki, M. S. Toledo, H. K. Takahashi, and A. H. Straus, Glycobiology 7:463-468, 1997), was used to detect GIPL-1 in Leishmania by indirect immunofluorescence and to analyze its role in macrophage infectivity. L. major promastigotes showed intense fluorescence with MEST-1, and GIPL-1 was detected in both amastigote and promastigote forms by high-performance thin-layer chromatography immunostaining by using MEST-1. Delipidation of L. major promastigotes with isopropanol-hexane-water eliminated the MEST-1 reactivity, confirming that only GIPL-1 is recognized in either amastigotes or promastigotes of this species. The biological role of GIPL-1 in the ability of L. major to invade macrophages was studied by using either Fab fragments of MEST-1 or methylglycosides. Preincubation of parasites with Fab fragments reduced macrophage infectivity in about 80% of the promastigotes and 30% of the amastigotes. Preincubation of peritoneal macrophages with p-nitrophenyl-beta-galactofuranoside (10 mM) led to significant ( approximately 80%) inhibition of promastigote infectivity. These data suggest that a putative new receptor recognizing beta-D-Galf is associated with L. major macrophage infectivity and that GIPL-1 containing a terminal Galf residue is involved in the L. major-macrophage interaction.
Suzuki
Erika
E
Department of Biochemistry, Universidade Federal de São Paulo/Escola Paulista de Medicina, Brazil.
Tanaka
Ameria K
AK
Toledo
Marcos S
MS
Takahashi
Helio K
HK
Straus
Anita H
AH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000913:Antibodies, Protozoan / Q000276:immunology*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D019084:Fluorescent Antibody Technique, Indirect; D005690:Galactose; D006017:Glycolipids / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D018320:Leishmania major / Q000254:growth & development / Q000472:pathogenicity*; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology; D017737:Macrophages, Peritoneal / Q000469:parasitology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010716:Phosphatidylinositols / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D006017:Glycolipids; D007140:Immunoglobulin Fab Fragments; D010716:Phosphatidylinositols; D005690:Galactose