Submitted to: Great Lakes International Imaging and Flow Cytometry Association
Publication Type: Abstract Only
Publication Acceptance Date: October 12, 2001
Publication Date: N/A
Technical Abstract: The goal of this study was to develop an assay to concurrently examine phenotype, activation, proliferation, apoptosis and death of antigen- or mitogen-stimulated bovine lymphocytes. Peripheral blood mononuclear cells (PBMC) were isolated from either non-infected or Mycobacterium bovis-infected calves. Once isolated, PBMC were labeled with PKH67 dye and cultured with no stimulation, M. bovis purified protein derivative or pokeweed mitogen. Cells were evaluated for activation (CD62L, CD44, CD25), phenotype (CD4, CD8, gdTCR), apoptosis (Annexin V and 7AAD), viability (7AAD) and proliferation (PKH67). Upon stimulation the mean fluorescence intensity (mfi) of CD62L decreased whereas the mfi of CD44 and CD25 increased on CD4**+ cells within the live gate. In general, activation marker expression was negligible on Annexin V**+ cells. This assay allows for a more detailed determination of the cellular immune response of cattle.