International Mammalian Genome Society

17th International Mouse Genome Conference

9-12 November 2003, Braunschweig, Germany


POSTER 194 - EIGHT FLUORESCENCE ANALYSIS ON A FOUR FLUORESCENCE FLOW ANALYSER

Deiters U
German Research Centre for Biotechnology, Braunschweig, Germany

Co-Authors: Mueller W
Institutions: German Research Centre for Biotechnology, Braunschweig, Germany

The work is a part of the EUMORPHIA project, dealing with the systemic analysis of the susceptibility of different mice inbred strains and mice mutants towards bacterial pathogens. For this, standardized operation procedures should be developed including characterization of the Ig profile or analysis of peripheral blood cells in mice of different strain, sex, and before and after infection. For the analysis of lymphocyte populations, either from pheripheral blood or lymphoid organs, we developed a robust flow cytometric method. The method uses a single staining procedure containing a mixture of eight directly coupled fluorochrome labeled antibodies against CD3 (Cychrome), CD4 (PE), CD8 (FITC), CD19 (APC), IgM (PE), IgD (FITC), F4/80 (PE) or CD11c (PE), and CD49b (FITC). It allows the determination of mature and immature B cells, CD4 and CD8 T cells, NK cells, DC, and macrophages. Using this method we analysed pheripheral blood of four inbred mouse strains (CBA/JHsd, C3H/HeN, BALB/c, C57BL/6J), male as well as female. It became apparent that there is a clear difference in the percentages of the various subpopulations between these strains. In addition, we observed differences in the subpopulations in male versus female mice, most obviously in the CBA/JHsd strain. We are now in the process of crossing the CBA/JHsd to C3H/HeN in order to identify genes which may control for the observed difference in the subpopulations between male and female CBA/JHsd mice. Supported by EUMORPHA, WP 6


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