Yang T J, Rabinovsky E D
Vet Immunol Immunopathol. 1987 Jan;14(1):77-84. doi: 10.1016/0165-2427(87)90076-6.
Various methods for separation of lymphocyte populations have been modified and adapted for use in isolating and identifying bovine lymphocytes. Ficoll diatrizoate (F-D) with a specific density of 1.084 was found to be superior to those with densities of 1.080 and 1.077 which were developed originally for the mouse and human mononuclear cells, respectively. F-D with a density of 1.084 attained a lymphocyte (absolute number) recovery rate of 92% whereas those with densities of 1.080 and 1.077 yielded 81% and 71% recovery rate of lymphocytes, respectively. Subsequent separation of T lymphocytes was achieved best by nylon wool column whereas separation of B lymphocytes was attained best by complement-mediated depletion of T lymphocytes with the T lymphocyte specific monoclonal antibody (MAb), BLT-1. The former yielded 95 +/- 3% T lymphocytes with 47 +/- 9% recovery rate, and the latter gave 96 +/- 3% B lymphocytes with 71 +/- 9% recovery rate. In comparison, direct panning of F-D gradient separated mononuclear cells with goat anti-bovine IgG coated plates yielded 80% B lymphocytes with 31% recovery rate and indirect panning of MAb BLT-1 treated F-D gradient-separated mononuclear cells with goat anti-mouse IgG coated plates yielded 89% T lymphocytes with 35% recovery rate.
多种分离淋巴细胞群体的方法已被改进并适用于分离和鉴定牛淋巴细胞。发现特定密度为1.084的泛影葡胺（F-D）优于最初分别为小鼠和人单核细胞开发的密度为1.080和1.077的泛影葡胺。密度为1.084的F-D的淋巴细胞（绝对数量）回收率达到92%，而密度为1.080和1.077的泛影葡胺的淋巴细胞回收率分别为81%和71%。随后，用尼龙毛柱能最好地实现T淋巴细胞的分离，而用T淋巴细胞特异性单克隆抗体（MAb）BLT-1通过补体介导的T淋巴细胞耗竭能最好地实现B淋巴细胞的分离。前者产生了95±3%的T淋巴细胞，回收率为47±9%，后者产生了96±3%的B淋巴细胞，回收率为71±9%。相比之下，用包被有山羊抗牛IgG的平板对F-D梯度分离的单核细胞进行直接淘选产生了80%的B淋巴细胞，回收率为31%，用包被有山羊抗小鼠IgG的平板对经MAb BLT-1处理的F-D梯度分离的单核细胞进行间接淘选产生了89%的T淋巴细胞，回收率为35%。
