Study of Fc gamma murine macrophage receptors (Fc gamma R) in the internalization of Histoplasma capsulatum yeasts.
Mária Lucía Taylor, B Rico, Federico M. Goodsaid
Abstract
Mária Lucía Taylor, B Rico, Federico M. Goodsaid
Abstract
Reports about Fc gamma R involvement in the phagocytosis of Histoplasma capsulatum yeast are contradictory. Fungal cells were incubated with anti-Histoplasma murine IgG or normal IgG as control. The opsonized yeasts were adjusted and spread on a J774.2 macrophage cell line monolayer. Yeast cells were allowed to attach to the monolayers at 4 degrees C for 1 h prior to initiating the internalization step of the phagocytic process, by shifting the cells to 37 degrees C. The participation of Fc gamma R in the internalization of yeast was monitored by radioiodination labelling of the macrophage surface proteins, and rosette-forming cells for erythrocytes coated with antibodies (RFC-EA). Results showed decreased [125I]-incorporation on the macrophage surface proteins at 15 min post-anti-Histoplasma IgG opsonized yeast infection. The same was observed when high and low virulent H. capsulatum strains were used. A decrease of RFC percentage was also observed at 15 min after IgG opsonized yeast phagocytosis. Results support the disappearance of Fc gamma R at 15 min from the surface of the macrophage that had ingested anti-Histoplasma IgG opsonized yeasts. The recovery of [125I]-counts and the RFC response at a longer time of phagocytosis suggest a possible Fc gamma R recycling mechanism.
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Reports about Fc gamma R involvement in the phagocytosis of Histoplasma capsulatum yeast are contradictory. Fungal cells were incubated with anti-Histoplasma murine IgG or normal IgG as control. The opsonized yeasts were adjusted and spread on a J774.2 macrophage cell line monolayer. Yeast cells were allowed to attach to the monolayers at 4 degrees C for 1 h prior to initiating the internalization step of the phagocytic process, by shifting the cells to 37 degrees C. The participation of Fc gamma R in the internalization of yeast was monitored by radioiodination labelling of the macrophage surface proteins, and rosette-forming cells for erythrocytes coated with antibodies (RFC-EA). Results showed decreased [125I]-incorporation on the macrophage surface proteins at 15 min post-anti-Histoplasma IgG opsonized yeast infection. The same was observed when high and low virulent H. capsulatum strains were used. A decrease of RFC percentage was also observed at 15 min after IgG opsonized yeast phagocytosis. Results support the disappearance of Fc gamma R at 15 min from the surface of the macrophage that had ingested anti-Histoplasma IgG opsonized yeasts. The recovery of [125I]-counts and the RFC response at a longer time of phagocytosis suggest a possible Fc gamma R recycling mechanism.
Key concepts: Internalization, Phagocytosis, Opsonin, Histoplasma, Macrophage, Antibody opsonization, Microbiology, Biology