Amino acid use due to activation of chicken lymphocyte subsets

Abstract

Previous research has shown that an appropriate balance of amino acids is required for cell proliferation and secretion of effector molecules by lymphocytes. However, quantitative changes in the requirements of lymphocytes for amino acids due to activation and differences between lymphocyte subsets are not known. Thus we conducted a series of experiments to identify the changes in protein and amino acid levels in resting and activated lymphocytes. Lymphocytes from adult male chickens were isolated from peripheral blood using a slow speed centrifugation followed by magnetic bead separation of subsets. Lymphocyte subsets were cultured in RPMI 1640 media supplemented with 0.1% chicken serum and stimulated with 15 μg/ml pokeweed mitogen (PWM) or no mitogen. After 72 hrs, cells were collected and total protein and amino acid levels were determined. Cell proliferation was confirmed by BrdU incorporation after 24 hrs. Results showed a significant increase in total protein per cell in stimulated CD4 cells compared to non‐stimulated cells (P<0.05). In addition, nonessential and essential amino acid levels were significantly increased in stimulated CD4 cells compared to non‐stimulated cells (P<0.05). However, Bu‐1 cells showed no significant difference in total protein per cell or levels due to stimulation. These data indicate amino acid use changes upon lymphocyte activation and is subset dependent. Supported by USDA Regional Research project 1013.