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Title: CHARACTERIZATION OF A SPONTANEOUSLY TRANSFORMED CHICKEN MACROPHAGE/MONOCYTIC CELL LINE HTC

Author
item RATH, N - USDA, ARS, PPPSR
item XIE, H - POULTRY SCIENCE CENTER
item PARCELLS, M - POULTRY SCIENCE CENTER
item SANTIN, E - POULTRY SCIENCE CENTER
item OKIMOTO, RON - POULTRY SCIENCE CENTER
item Lillehoj, Hyun

Submitted to: Poultry Science Association Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 5/1/2002
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Transformed and immortalized cells provide excellent models to study many functional aspects of their naturally occurring counterparts because of the ease of manipulations and their abilities to undergo innumerable cell divisions. Such cells are generated naturally or in culture through neoplastic transformation induced by viruses, chemicals, and genetic rearrangements. In the process of isolating and culturing chicken heterophils (HT) from peripheral blood, a flask of cells was inadvertently left in the incubator over a period of 3 weeks during which time an island of cells arose which exhibited some what an epithelial-liketype morphology. We expanded these cells in RPMI-1640 medium containing 5% fetal bovine serum (FBS) and stained to ascertain whether these belong to HT-granulocyte class by staining with fluorescein isothiocynate (Rath et al, 1998) and found those not to be granulocytes. We have subsequently grown these cells in media supplemented with 1-2% chicken serum alone and have successfully passaged more than 70 times. Although attaching to the flasks, these cells tend to bud off and grow as aggregates. To characterize further, we stained these cells (referred to as HTC hereafter) with a panel of monoclonal antibodies raised against several chicken specific cell surface antigens and performed flow cytometry. The HTC cells were positive to the following markers, MHC class II, K1, K55, CD44, CD164-sialomucin, suggestive of macrophage/monocyte lineage. HTC cells also responded to a number of stimuli including lipopolysaccharide (LPS), recombinant chicken interferon-g, phorbol myristate acetate, and zymosan, and produced interleukin-6, nitrite, hydrogen peroxide. LPS stimulated production of two matrix metalloprpteinase (MMP) bands corresponding to MW ~ 59 kDa, and ~150 kDa. The phagocytic activity of HTC cells was also assessed using latex bead uptake. Using specific primer pairs for Avian Leukosis Virus (ALV), Marek=s disease virus, reticuloendothelial virus, herpes virus of turkeys, and polymerase chain reaction (PCR) amplification, the cells were positive for ALV only. Similarly, these cells were positive for IFN-g, IL-1-b, TGF-b4, IL-6, and MMP-2 using chicken specific primers. Our results show the HTC cells have macrophage/monocyte characteristics, and may be useful in the study of cytokine regulation and immunomodulations in poultry.