|Green, Benedict - Ben|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 9/22/2008
Publication Date: 11/1/2008
Citation: Chitko Mckown, C.G., Chapes, S.K., Miller, L.C., Green, B.T. 2008. Characterization of the porcine monocyte-derived cell lines Cdelta2+ and Cdelta2+ [abstract]. Society for Leukocyte Biology. Poster No. 45.
Technical Abstract: Cell lines Cdelta2- and Cdelta2+ were developed from monocytes obtained from a 10-month-old, crossbred, female pig at the U.S. Meat Animal Research Center, Clay Center, NE. These cells have macrophage morphology, stain positively for alpha-naphthyl esterase and negatively for peroxidase. Additionally, these cell lines are bactericidal and express iNOS in response to treatment with LPS (Chitko-McKown et al., Proceedings of the Conference of Research Workers in Animal Disease, 2004). In order to more fully characterize these cell lines, additional experiments were performed. Cytokine mRNA expression was measured in LPS-treated cells by means of qRT-PCR using three nested primer sets with probes specific for lymphokines, monokines, and house-keeping genes, respectively. The cell lines expressed the cytokines IL-1alpha, IL-6 and IL-10, but not IL-2 and IL-4. Phagocytosis was measured with fluorescent latex beads followed by flow cytometric quantitation. The cell lines were highly phagocytic. By 18 h, over 85% of the Cdelta2- cells had engulfed latex beads as compared to over 97% of the Cdelta2+ cells. Cell-surface expression of monocyte CD antigens was measured by flow cytometry using antibodies specific for porcine CD antigens. Both cell lines expressed the porcine cell surface molecules MHCI, CD11b, CD14, CD16, CD172, and small amounts of CD2. The lines were negative for the mouse marker H2K**k and the bovine control. Additionally, cells were tested for the presence of Bovine Viral Diarrhea Virus and Porcine Circovirus Type 2, and were negative for both. These cells resemble porcine macrophages in morphology and activity and will be useful tools in laboratories studying porcine macrophage biology. Supported in part by USDA-ARS Project Numbers 5438-32000-029-00D and 3625-32000-088-00D; NASA Grant NAG2-1274; NIH Grants AI55052, AI052206, RR16475, and RR17686; and the Kansas Agricultural Experiment Station.