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Cell Sorting for Functional and Post-Genomic Analysis of Rare Immune and Infected Cells

Institutions
Institute for Animal Health
Start date
2005
End date
2006
Objective
This application is to purchase a state of the art Fluorescent Activated Cell Sorter (FACS). The FACSAria will enable us to purify rare immune or infected cells in multicolour stained samples of blood or tissues. It will enable the cells to be isolated in sufficient numbers and sufficiently rapidly to allow functional studies by immunological methods and proteomic and transcriptomic analyses.

The key cells that are contributing to and affecting the intensity and bias of the immune response or are central to pathogenesis are frequently present in very low numbers and are only identifiable following multicolour staining with panels of monoclonal antibodies. Thus, their identification and purification is only possible by flow- cytometric sorting. Such cells include dendritic cells, NK cells and virus infected cells.

The studies will be in the natural target species and will exploit a unique combination of facilities at IAH, namely the availability of families and lines of MHC defined inbred cattle and chickens, the facility for surgical cannulation to provide afferent lymph for isolating dendritic cells, cellular immunological and post-genomic analyses employing transcriptomics and proteomics and disease models in the natural target species.

The diseases that the studies will be applied to in the first instance include bovine tuberculosis and foot and mouth disease in cattle. In chickens Mareks disease and infections with Eimeria and Salmonella will be the initial focus. The interaction of these organisms with dendritic cell sub-populations, the stimulation of NK cell responses and the responses of T cells, in particular memory responses to vaccination, will be major themes of the research programs. Infection of cells by Mareks disease virus and viral gene expression in relation to neoplastic transformation will also be a primary objective. The vaccinology programmes will focus on the induction of memory using peptide MHC tetramers to identify immune cells in the context of the MHC-defined animals noted above.

The studies are aimed at developing means for the control of infectious disease by making use of emerging technology, namely transcriptomics and proteomics, and to integrate cellular immunology and post-genomic studies.

Funding Source
Biotechnology and Biological Sciences Research Council
Project number
BB/C511256/1
Categories
Salmonella
Bacterial Pathogens
Commodities
Meat, Poultry, Game