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Induction of novel protein synthesis by opsonizedHistoplasma capsulatum ingested by murine peritoneal macrophages
Authors:Katsuhiko Kamei  Elmer Brummer  Karl V. Clemons  Dr David A. Stevens
Affiliation:(1) Division of Infectious Diseases, Department of Medicine, Santa Clara Valley Medical Center, and California Institute for Medical Research, San Jose, California;(2) Stanford University School of Medicine, Stanford, California, USA;(3) Division of Infectious Diseases, Department of Medicine, Santa Clara Valley Medical Center, 751 South Bascom Avenue, 95128-2699 San Jose, CA, USA
Abstract:It is known thatHistoplasma capsulatum can resist the intraphagolysosomal environment and multiply inside macrophages. This resistance can be closly related to its pathogenicity. The mechanism of this resistance has been investigated, but it has not been clarified as yet. To learn about the metabolic condition of the yeast-form ofH. capsulatum (isolates G217B and CDC 105) when ingested by macrophages, we investigated protein synthesis by ingestedH. capsulatum with [35S]-methionine labeling. Cycloheximide at 5 to 10 µg/ml was used to preferentially inhibit macrophage uptake of [35S]-methionine without affectingH. capsulatum uptake. Protein synthesis byH. capsulatum in medium alone served as a positive control. The negative control consisted of macrophages with ingested heat-killedH. capsulatum. Analysis of cytosols with SDS-PAGE and fluorography disclosed that, respectively for G217B and CDC 105, ingestedH. capsulatum synthesized 4 and 5 novel proteins, increased the synthesis of 9 and 17 proteins and decreased the synthesis of 9 and 10 constitutive proteins. Ten of these novel or increased proteins were apparently common to both strains. These metabolic changes in ingestedH. capsulatum could reflect its adaptation to the intraphagolysosomal environment of macrophages and its ability to multiply there.
Keywords:H. capsulatum  Macrophages  Novel proteins
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