Williams D L, Lo J L, Amborski G F
Vet Immunol Immunopathol. 1986 Feb;11(2):199-204. doi: 10.1016/0165-2427(86)90098-x.
A simple and efficient method to enrich bovine T lymphocytes from peripheral blood mononuclear cells (PBMC) by immuno-affinity depletion ("panning") has been developed. The PBMC were initially separated by density gradient centrifugation on Histopaque of density 1.077 g/ml. The T lymphocyte subset was then separated from PBMC by depletion of membrane immunoglobulin (Ig) bearing cells which had an affinity for anti-Ig antibodies bound to polystyrene tissue culture flasks. An average of 95% of the nonadherent "panned" cells were identified as T lymphocytes using a label of peanut agglutinin conjugated with fluorescein isothiocyanate (PNA-FITC). Two percent of the PNA negative cells were Ig bearing cells. The average yield was 50% of the original T lymphocytes found in the PBMC population, and the cell viability as assessed by trypan blue exclusion was greater than 95%. The separation took approximately 2 hours, and the total number of T lymphocytes recovered from 40 ml of blood was in the range of 20-40 X 10(6).
已开发出一种通过免疫亲和去除法（“淘选”）从外周血单核细胞（PBMC）中富集牛T淋巴细胞的简单高效方法。PBMC最初通过在密度为1.077 g/ml的Histopaque上进行密度梯度离心分离。然后通过去除与结合在聚苯乙烯组织培养瓶上的抗Ig抗体具有亲和力的膜免疫球蛋白（Ig）携带细胞，从PBMC中分离出T淋巴细胞亚群。使用与异硫氰酸荧光素偶联的花生凝集素（PNA-FITC）标记，平均95%的非贴壁“淘选”细胞被鉴定为T淋巴细胞。2%的PNA阴性细胞是Ig携带细胞。平均产量为PBMC群体中原始T淋巴细胞的50%，通过台盼蓝拒染法评估的细胞活力大于95%。分离过程约需2小时，从40 ml血液中回收的T淋巴细胞总数在20 - 40×10⁶范围内。
