Research Project
10171426
SUMMARY The Core C facility supports research addressing the specific aims of the projects to include the planning and execution of experimental sepsis cohort studies, hematological analyses of whole blood samples, and measurements of multiple circulating glycoproteins that includes comparative studies of markers of inflammatory processes during the onset and progression of sepsis. The Core thereby provides support addressing the central hypothesis of the program, Protein glycosylation and glycoprotein remodeling alter the coagulopathy and inflammation of sepsis. Core C provides essential experimental and technological capabilities that include the analysis of experimental and clinical biological samples derived from mouse and human species including whole blood, serum, plasma, and, when requested, urine. In collaboration with the projects, and as published, the core has developed an experimental sepsis protocol in the mouse that includes standardized and reproducible pathogen infection and monitoring with specific thresholds for data inclusion representing the onset and progression of mouse sepsis in response to five different human bacterial pathogens and clinical isolates. This approach has enabled comparative studies spanning the specific aims of each of the three projects while revealing the presence of different pathogenic pathways that provide a degree of stratification in the pathogenesis of sepsis due to specific bacterial pathogens. These studies will be further expanded as indicated in the research projects of this application. In addition, the core facility will complete hematology analyses of mouse blood samples as well as blood from human volunteers and sepsis patients to include measurements of white and red blood cells, hematocrit values, hemoglobin abundance, and multiple platelet metrics. Additional analyses when requested will include flow cytometric studies of cell abundance representing various blood cell lineages including T and B lymphocytes, neutrophils, monocytes, eosinophils, and basophils. These hematology analyses will be provided to onsite local project researchers and will be standardized with identical equipment that is used to undertake comparable hematology analyses at the second site of project research. The core facility will also generate multiplex analyte datasets from serum, plasma, or urine samples from mouse and human species in the context of health and sepsis. These multiplex experiments use investigator-selected and customizable assays yielding quantitative measurements of essential and important cytokines, growth factors, and various other biological indicators of physiological systems especially those that impinge upon inflammatory processes. Over a thousand proteins can be selectively measured in various sets of analytes, and many of these proteins are found at low abundance and cannot be routinely detected and accurately measured using mass spectrometry. The core thereby provides a means to query lower abundance proteins that are difficult to quantify otherwise. Core C will enhance the rate of progress of program research and facilitate research integration and synergies among the studies proposed.
