title: Demonstration of leptospiral antigens on tissues using monoclonal antibodies and avidin-biotin peroxidase staining.
abstract: Glycolipoprotein (GLP) cytotoxin was extracted from Leptospira interrogans serovar canicola. The silver staining profile of GLP subjected to SDS-PAGE under denaturing conditions showed a number of bands in the mol. weight range of 14-66 kDa. Mouse Monoclonal Antibodies (MAbs) IgG3 recognizing a band near to 24 kDa of leptospiral GLP were produced (clone number MGLP-01). The agglutinating property of MAbs was established by microscopic agglutination test (MAT) using 25 different serovars as antigens. Only the homologous serovar was agglutinated by MAbs suggesting that the recognized epitope is a specific surface-exposed antigen. The MAbs were applied to demonstration of leptospiral antigens in tissue damage by avidin-biotin immunoperoxidase staining. Golden hamsters were experimentally infected with a virulent strain of L. interrogans serovar canicola. Histologically kidneys stained by routine hematoxylin and eosin showed changes characterized by injury of tubular epithelial cells leading to acute tubular necrosis (ATN). Typical, well-defined morphologic leptospires or finely granular deposits were found by immunoperoxidase staining near to blood vessels, within inflammatory infiltrates and intraluminal in proximal and distal parts of the nephron. Binding of leptospiral antigens to capillary endothelial cells, tubular epithelial cells and macrophages were also demonstrated. This entails a basis for further studies either in research or in diagnostic histopathology.
authors.affiliation: Department of Bacteriology, Oswaldo Cruz Institute-IOC/FIOCRUZ, Brasil.
authors.forename: M M
authors.initials: MM
authors.lastname: Pereira
authors.forename: J
authors.initials: J
authors.lastname: Andrade
authors.forename: M D
authors.initials: MD
authors.lastname: Lacerda
authors.forename: N M
authors.initials: NM
authors.lastname: Batoréu
authors.forename: R S
authors.initials: RS
authors.lastname: Marchevsky
authors.forename: R
authors.initials: R
authors.lastname: Ribeiro dos Santos
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000942:Antigens, Bacterial / Q000032:analysis*; D001426:Bacterial Proteins / Q000302:isolation & purification; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas; D007124:Immunoenzyme Techniques; D007683:Kidney Tubular Necrosis, Acute / Q000382:microbiology* / Q000473:pathology; D007684:Kidney Tubules / Q000382:microbiology; D007921:Leptospira interrogans / Q000276:immunology*; D008647:Mesocricetus; D051379:Mice; D008810:Mice, Inbred C57BL; D014895:Weil Disease / Q000382:microbiology* / Q000473:pathology
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins