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ARS Home » Southeast Area » Fayetteville, Arkansas » Poultry Production and Product Safety Research » Research » Publications at this Location » Publication #315667
Title: Proteomic analysis of macrophage activated with salmonella lipopolysaccharide

Author
item MAKKAR, SARBJEET - University Of Arkansas
item LIYANAGE, ROHANA - University Of Arkansas
item LAY, JACKSON - University Of Arkansas
item Rath, Narayan

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 3/11/2015
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Macrophages play pivotal role in immunity. They are activated by many pathogen derived molecules such as lipopolysaccharides (LPS) which trigger the production of various proteins and peptides that drive and resolve inflammation. There are numerous studies on the effect of LPS at the genome level but knowledge of its proteomics-related effects is limited. To determine the proteomic effect with LPS stimulation, we treated chicken macrophages HTC with Salmonella typhimurium LPS by using stable isotope labelling of amino acids in cell culture (SILAC). The heavy (H) (13C-lysine) labelled cells were treated with LPS and light (L) cells were treated as controls. After 16 hrs of stimulation, the protein was extracted, quantified and an equal amount from H and L cells was mixed, subjected to reduction/alkylation, and trypsin digestion followed by liquid chromatography/ tandem mass spectrometry. We quantified 133 proteins using Skyline software, out of which 13 were upregulated and 11 down regulated. Some of the upregulated proteins included HSP 70 and 90, DNA helicases, prostaglandin D synthase, and CCL5 ligand. The down regulated proteins included ubiquitin, proteasome, and proteins related to protease activity. In conclusion this study suggests that activation of macrophages with LPS results in the differential expression of proteins involved in cytoskeleton remodeling, cell migration and cell signaling which thereby mediate the immune response.