title: Soluble components of Histoplasma capsulatum var. capsulatum have hemagglutinin activity and induce syngeneic hemophagocytosis in vitro.
abstract: Histoplasma capsulatum var. capsulatum is a thermally dimorphic fungus that causes histoplasmosis. Fungal hemagglutination activity and cases of reactive hemophagocytic syndrome (RHS) have been reported in the disseminated form of disease. In the present study, soluble components of H. capsulatum var. capsulatum have been investigated for hemagglutinin activity and the capacity to induce hemophagocytosis in the mouse system. To analyze hemagglutinating activity, mouse red blood cells (RBC) (1% v/v in PBS) were incubated (37 degrees C, 1 h) with cell-free antigen (CFAg) from H. capsulatum var. capsulatum (isolate IMT/HC128) (RBC-CFAg) or previously heated CFAg (56 degrees C, 30 min) (RBC-hCFAg) or as control with PBS (RBC-PBS). Hemophagocytosis was analyzed by incubating BALB/c mouse peritoneal phagocytic cells (5 x 10(6) cells) with syngeneic RBC, sensitized or not with CFAg. In addition, mouse polyclonal antibodies were raised against syngeneic RBC-CFAg (anti-RBC-CFAg) and used to analyze CFAg chromatographic fractions (Sephadex G75/120) by immunoenzymatic assay (ELISA). Hemagglutinin activity was observed with RBC-CFAg, but not with RBC-hCFAg or RBC. Also, hemophagocytosis was observed with RBC-CFAg, but not with RBC. The anti-RBC-CFAg antibodies reacted with CFAg fractions corresponding to a molecular mass (MM) higher than 150 kDa. In conclusion, the yeast form of H. capsulatum var. capsulatum releases thermolabile soluble components with hemagglutinin activity and it has been demonstrated for the first time that soluble components of the same fungus induce syngeneic hemophagocytosis in the in vitro mouse system. Also, indirect analysis with antibodies suggests that high-MM components (>150 kDa) are responsible for the interaction with RBC.
authors.affiliation: Departamento de Ciências Patológicas, CCB, Universidade Estadual de Londrina, Campus Universitário, Londrina, PR, 86051-970, Brazil.
authors.forename: R H F
authors.initials: RH
authors.lastname: Vivan
authors.forename: P C
authors.initials: PC
authors.lastname: Leonello
authors.forename: L A
authors.initials: LA
authors.lastname: Nagashima
authors.forename: M S
authors.initials: MS
authors.lastname: Kaminami
authors.forename: F S M
authors.initials: FS
authors.lastname: Tristão
authors.forename: A
authors.initials: A
authors.lastname: Sano
authors.forename: M A
authors.initials: MA
authors.lastname: Ono
authors.forename: C V
authors.initials: CV
authors.lastname: Béjar
authors.forename: E N
authors.initials: EN
authors.lastname: Itano
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D005656:Fungal Proteins / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D006384:Hemagglutination; D006658:Histoplasma / Q000737:chemistry*; D017737:Macrophages, Peritoneal / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D010587:Phagocytosis
chemical_list: D005656:Fungal Proteins