The In Vitro Core will provide the centralized bacterial, cell culture and molecular microbiology diagnostic support needed for each of the Projects. Cp and Pg will be grown in high-volume at high titter for in-vivo inoculation experiments, and for in-vitro cell stimulation experiments. Purified Cp will be prepared in the appropriate cell lines to avoid cross species antigen stimulation. Cp and Pg will be purified at high level for in-vitro experiments. Cp fluorescence staining will be use to evidence infection for in-vitro experiments, and the morphology and maturation of Cp inclusions. The Core will culture macrophages from wild type and knockout mice, to be challenged with Cp and Pg for the study cytokine profiles and foam cell formation. DNA will be extracted from mice tissue (gingival, blood, lung, liver, spleen, and aorta), collected at different time points during infection, and microbial (Cp or Pg) DNA will be detected by PCR using primer sets specific for each microorganism. The number of organism (loads) contained in the tissue sample will be quantified using a new end point quantitative PCR method (a-PCR) that we have previously validated to study Ct infection and transmission. This assay has an excellent correlation with quantitative Ct culture. Cp and Pg loads will be quantified in in-vivo experiments. The presence of a threshold in the number of organisms in tissue that divide mice into responder vs. non-responders to bacterial antigen stimulation will be investigated. We hypothesize that the innate response is dependent on the organism load, and that un-infected mice and mice with low organism loads will have a lower response (or no response) compared with mice having higher organisms loads. We will study how organisms loads: 1) are modified by changes in innate immunity respond in infected gene-deficient mice (IL-p, IL-1R, ApoE, and LXR knockout mice);2) influence the acceleration of atherosclerosis in mice infected with either Pg or Cp;and 3) influence oral bone loss in mice infected with Pg.
Common bacterial pathogens such as Chlamydia pneumoniae (Cp) which often cause unrecognized upper respiratory infection, and Porphyromonas gingivalis (Pg) the main cause of periodontal disease, could also cause long lasting relentless inflammatory illnesses that result in the formation of intra-vascular lesions and in the case of Pg bone loss. The studies in this proposal seek to gain a better understanding of how these pathogens adapt to colonize humans while evading the natural immune defenses, and the long lasting inflammatory processes they induce. The In Vitro Core will provide the molecular and diagnostic support for all the projects in this proposal.
|Papadopoulos, G; Shaik-Dasthagirisaheb, Y B; Huang, N et al. (2016) Immunologic environment influences macrophage response to Porphyromonas gingivalis. Mol Oral Microbiol :|
|Shaik-Dasthagirisaheb, Yazdani B; Mekasha, Samrawit; He, Xianbao et al. (2016) Signaling events in pathogen-induced macrophage foam cell formation. Pathog Dis 74:|
|He, Xianbao; Liang, Yanmei; LaValley, Michael P et al. (2015) Comparative analysis of the growth and biological activity of a respiratory and atheroma isolate of Chlamydia pneumoniae reveals strain-dependent differences in inflammatory activity and innate immune evasion. BMC Microbiol 15:228|
|Beaulieu, Lea M; Clancy, Lauren; Tanriverdi, Kahraman et al. (2015) Specific Inflammatory Stimuli Lead to Distinct Platelet Responses in Mice and Humans. PLoS One 10:e0131688|
|Koupenova, Milka; Mick, Eric; Mikhalev, Ekaterina et al. (2015) Sex differences in platelet toll-like receptors and their association with cardiovascular risk factors. Arterioscler Thromb Vasc Biol 35:1030-7|
|Huang, N; Shaik-Dasthagirisaheb, Y B; LaValley, M P et al. (2015) Liver X receptors contribute to periodontal pathogen-elicited inflammation and oral bone loss. Mol Oral Microbiol 30:438-50|
|Shaik-Dasthagirisaheb, Y B; Huang, N; Weinberg, E O et al. (2015) Aging and contribution of MyD88 and TRIF to expression of TLR pathway-associated genes following stimulation with Porphyromonas gingivalis. J Periodontal Res 50:89-102|
|Shaik-Dasthagirisaheb, Yazdani B; Huang, Nasi; Gibson 3rd, Frank C (2014) Inflammatory response to Porphyromonas gingivalis partially requires interferon regulatory factor (IRF) 3. Innate Immun 20:312-9|
|Koupenova, Milka; Vitseva, Olga; MacKay, Christopher R et al. (2014) Platelet-TLR7 mediates host survival and platelet count during viral infection in the absence of platelet-dependent thrombosis. Blood 124:791-802|
|Kramer, Carolyn D; Weinberg, Ellen O; Gower, Adam C et al. (2014) Distinct gene signatures in aortic tissue from ApoE-/- mice exposed to pathogens or Western diet. BMC Genomics 15:1176|
Showing the most recent 10 out of 27 publications