, 2011). In Histoplasma, only a handful of factors have been demonstrated to contribute to virulence

in vitro or in vivo, and even fewer have been tested for virulence roles in both strain backgrounds. In the following sections, we will discuss studies in G186A and G217B as Torin 1 representative for the Panamanian and NAm2 phylogenetic clades, respectively. The secreted protein Cbp1 was the first Histoplasma virulence factor to be established through genetics. Both G217B and G186A yeast cells produce abundant Cbp1 during liquid culture (Kugler et al., 2000; Youseff et al., 2009), and the CBP1 gene is expressed by both strains during intramacrophage growth and during in vivo infection (Batanghari et al., 1998; Edwards et al., 2011). Cbp1 is required for the full virulence

of G186A and G217B. Genetic mutations for proof of this were provided through the creation of a cbp1-deletion allele in the G186A background (Sebghati et al., http://www.selleckchem.com/products/pirfenidone.html 2000) and isolation of a T-DNA insertion mutant in the CBP1 gene in the G217B background that prevents Cbp1 production (Youseff et al., 2009). In the absence of Cbp1, Histoplasma yeast grow at a similar rate in culture; however, the yeast are attenuated in both macrophage and mouse assays of virulence (Sebghati et al., 2000; Edwards et al., 2011). While the exact mechanism of Cbp1 contribution to virulence remains unknown, the Cbp1 homodimer has structural similarity to mammalian saposin B (Beck et al., 2009) suggesting a role in transforming the phagocytic compartment into a permissive environment for yeast survival and replication. The Cbp1 requirement for both G186A and G217B virulence indicates conservation of at least one mechanism for pathogenesis. G186A and G217B yeast cells have similar size and morphology when viewed by light microscopy, however, structural and chemical differences exist between their respective cell walls. Electron microscopy shows that the cell wall of G186A is more than twice as thick as the cell wall of G217B (Edwards

Tyrosine-protein kinase BLK et al., 2011). Biochemical analysis of the cell walls following sodium hydroxide or glucanase treatment classifies strains as one of two chemotypes based on the polysaccharide composition of the yeast cell wall (Domer, 1971; Kanetsuna et al., 1974; Reiss, 1977; Reiss et al., 1977). Chemotype II comprise those strains for which the yeast cell wall contains α-glucan whereas Chemotype I strains lack α-glucan in the yeast cell wall. Follow-up studies using immunogold labeling confirmed the presence of α-glucan in the yeast cell walls of Chemotype II strains G186A (Panamanian class) and UCLA531 (a North American isolate with the same restriction fragment length polymorphism pattern and fatty acid profile as the Downs NAm1 strain) (Eissenberg et al., 1997; Zarnowski et al., 2007b). In contrast, the NAm2 strain G217B lacks α-glucan defining it as Chemotype I (Eissenberg et al., 1991).