Dataset Information

Dynamics of Increasing IFN-? Exposure on Murine MH-S Cell-Line Alveolar Macrophage Phagocytosis of Streptococcus pneumoniae.

ABSTRACT: Previous investigations have demonstrated that activation with the type II interferon, IFN-?, downregulates alveolar macrophage (AM) phagocytosis of Streptococcus pneumoniae. While these studies have shown clear effects at discrete time points, the kinetics of the macrophage response to IFN-? over time, with respect to pneumococcal phagocytosis, have not been shown. Here, we describe these kinetics in the murine MH-S AM cell-line, a well-established model useful for investigations of AM phenotype and function. We measure binding and internalizing rates of S. pneumoniae following exposure to increasing durations of physiologic levels of IFN-?. When MH-S murine alveolar macrophage (mAM) were exposed to IFN-? for increasing durations of time, from 0 to 6 days before inoculation with the type II S. pneumoniae, D39, exposure for 6?h transiently reduced bacterial binding by 50%, which was temporarily restored at 2 and 3 days of exposure. Bacterial internalization was also reduced shortly following initial exposure, however, internalization continued to fall to less than 5% that of IFN-? naïve controls after 6 days of exposure. These data may help explain otherwise contradictory reports from the literature regarding timing between infections and reductions in macrophage function.

SUBMITTER: Mina MJ

PROVIDER: S-EPMC4692125 | biostudies-literature | 2015 Jun

REPOSITORIES: biostudies-literature

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Dynamics of Increasing IFN-γ Exposure on Murine MH-S Cell-Line Alveolar Macrophage Phagocytosis of Streptococcus pneumoniae.

Mina Michael J MJ Brown Lou Ann S LA Klugman Keith P KP

Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research 20150225 6

Previous investigations have demonstrated that activation with the type II interferon, IFN-γ, downregulates alveolar macrophage (AM) phagocytosis of Streptococcus pneumoniae. While these studies have shown clear effects at discrete time points, the kinetics of the macrophage response to IFN-γ over time, with respect to pneumococcal phagocytosis, have not been shown. Here, we describe these kinetics in the murine MH-S AM cell-line, a well-established model useful for investigations of AM phenotyp ...[more]

PMID: 25713979

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Project description:During the infection process, both the host and the pathogen undergo a dynamic cascade of events that result in altered gene expression patterns (Westermann et al. 2012). With dual-RNA analysis, the complex interaction between a pathogen and its host during infection can be revealed simultaneously at their RNA levels (Westermann et al. 2012). Therefore, dual-transcriptomic analysis on JMRC:FSU:09682 and MH-S was performed to understand the pathogenesis at the genetic level to identify what kind of stresses that L. corymbifera encounter as well as the transcriptional response of macrophages during the infection. L. corymbifera (JRMC:FSU:09682) was cultivated for 7 days in KK1 medium (Kraibooj et al. 2014) at 37 °C . Murine alveolar MH-S macrophages (ATCC: CRL-2019) were cultivated in RPMI-1640 supplemented with 10% heat inactivated fetal bovine serum at 37 °C in 5 % CO2. Macrophages were seeded in 6 well plates at 106 cells per well to adhere overnight. Macrophages were infected with spores at a MOI (Multiplicity of Infection) of 5. After 3 hr of co-incubation with MH-S, extracellular spores were removed by at least 3 washing steps with pre-warmed RPMI-1640 (PAA Laboratories) and incubated for 13 hr in prior to the RNA extraction procedure. Controls in these experiments were fungus grown for 16 hours without macrophages and macrophages without fungal spores. Fungal RNA was isolated using RiboPure Yeast Kit (Thermo Fisher) and macrophage RNA was isolated with the RNeasy Mini kit (Qiagen), according to the manufacturers’ instructions. The concentration and purity of RNA samples were assessed using a NanoDrop spectrophotometer and Agilent 2100 bioanalyzer. A total of 681 L. corymbifera genes were differentially expressed during the co-infection with macrophages, with 629 down- (data not shown) and 52 up-regulated (Table 4.1). In order to systematically analyse the function of the differentially expressed genes (DEG), GO enrichment analyses were performed using FungiFun2.

Dataset Information

Dynamics of Increasing IFN-? Exposure on Murine MH-S Cell-Line Alveolar Macrophage Phagocytosis of Streptococcus pneumoniae.

Publications

Dynamics of Increasing IFN-γ Exposure on Murine MH-S Cell-Line Alveolar Macrophage Phagocytosis of Streptococcus pneumoniae.

Similar Datasets

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