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Real time quantitative PCR analysis of murine macrophages

ABSTRACT: Murine macrophages were isolated from the lungs of mice given a pulmonary challenge with C. neoformans strain H99. Mice were either given a protective (H99γ) or a mock (HKCn) immunization prior to C. neoformans H99 challenge, and macrophages were isolated from the lungs of mice 24 hours, 3 days, or 7 days post-challenge using anti-CD11b microbeads according to the Miltenyi cell sorting system. We used SA Biosciences Toll-like Receptor PCR assay panel to quantitate gene expression of signal transduction factors in total RNA isolated from macrophages derived from immunized mice compared to non-immunized. qPCR gene expression profiling. Macrophages from 5 mice per group were pooled and assayed as indicated in the summary. Each experiment was performed 3 times and the resulting Ct values of each group from each experiment averaged prior to data analysis. TIme points were analyzed separately

ORGANISM(S): Mus musculus

SUBMITTER: Sarah Hardison 

PROVIDER: E-GEOD-39990 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Protective immunity against pulmonary cryptococcosis is associated with STAT1-mediated classical macrophage activation.

Hardison Sarah E SE   Herrera Gina G   Young Mattie L ML   Hole Camaron R CR   Wozniak Karen L KL   Wormley Floyd L FL  

Journal of immunology (Baltimore, Md. : 1950) 20120914 8

Experimental pulmonary Cryptococcus neoformans infection in BALB/c mice is associated with polarized Th2-type cytokine production, alternative macrophage activation, and severe bronchopneumonia. In contrast, pulmonary infection with a C. neoformans strain that secretes IFN-γ, H99γ, elicits Th1-type cytokine production and classical macrophage activation. Additionally, mice infected with H99γ resolve the acute infection and are subsequently protected against challenge with wild-type C. neoformans  ...[more]

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