The Imaging Core will provide analytic procedures that evaluate the studies performed in Projects 1-3 to elucidate the pathologic events occurring during Coccidioides spp. infection. Visualizing the immune events and the host-pathogen interactions is critical to understanding the in vivo processes studied. The Imaging Core consists of four components: (1) immunohistochemistry/in situ hybridization; (2) confocal laser scanning microscopy; (3) flow cytometry; and (4) microscopic image capture and analysis. The objectives of the core are ? to characterize the histological and immunological immune response to Coccidioides spp. in intranasal infection through immunohistochemical staining of infected tissue, confocal laser scanning microscopic evaluation of lesion cells and cytokines, and analysis of microscopy results using image capture (Project 1, Specific Aims #1-3). The image capture component will interface with immunohistochemistry and confocal laser scanning microscopy techniques to provide digitalized image profiles for quantitative analysis of cell and cytokine populations within affected pulmonary tissue. ? to verify the in vitro gone expression patterns that are up-regulated during the parasitic phase of Coccidioides spp. in the murine model (Project 2, Specific Aim #2). ? to characterize via flow cytometry cell surface phenotype, function, and cytokine profiles from dendritic cells pulsed with candidate vaccine antigens, and from peripheral blood monocytes responding to the pulsed dendritic cells, (Project 3, Specific Aims #1-3).
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