Adherence as an in vitro assessment of neutrophil function in alpacas Public

http://ir.library.oregonstate.edu/concern/honors_college_theses/3197xp278

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  • Camelids are known for frequent occurrence of general ill-thrift. In other species, neutrophil dysfunction has been identified as a cause of ill-thrift, and linked to chronic fungal and bacterial infections. However, no reference values exist for neutrophil function testing in camelids, which inhibits accurate evaluation of neutrophil deficiency in this species. Neutrophil adhesion values were investigated in six healthy alpacas. Neutrophils were isolated from whole, EDTA-treated blood once per week for three weeks and suspended in RMPI 1560, supplemented with 1% heat-inactivated fetal bovine serum. Five hundred thousand cells were added to each well in a 96-well non-tissue culture treated plate and either untreated or stimulated with phorbol 12-myristate 13-acetate (PMA). Non-adherent cells were discarded, adhered cells were stained with crystal violet gram staining solution, and plates were washed with deionized water. Stain was solubilized with 1% SDS and percent adherence determined by absorbance at 570 nm. No significant difference was found in the percent neutrophil adherence between weeks (p = 0.59). Mean percent neutrophil adhesion was 12.4% ± 3.0% (9.4% – 15.4%). Given that results from individual alpacas appear to be relatively consistent at different time points, use of this adhesion assay may be a valuable tool in investigating neutrophil function in camelids.
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