Talbot M D, Jasani M K
J Immunol Methods. 1985 Nov 28;84(1-2):165-75. doi: 10.1016/0022-1759(85)90425-9.
Measurement of the activity of the enzyme DNA polymerase alpha has been investigated with regard to its potential usefulness as a method for the detection and quantification of lymphocyte activation in vivo. A modified enzyme assay was developed in order to optimise measurement of activity in crude homogenates of cells or tissues, thus allowing the convenient handling of multiple samples. Specificity of the assay for polymerase alpha was ensured by the inclusion in the assay mixture of dideoxythymidine triphosphate, an inhibitor of the other eukaryotic DNA polymerases. The activity of DNA polymerase alpha was found to be closely correlated with [3H]thymidine incorporation in a mitogen-stimulated in vitro system. The usefulness of the polymerase alpha method for the quantification of lymphocyte activation was validated in 3 different in vivo systems of either immune-mediated or drug-induced lymphoid cell response.
关于将DNA聚合酶α的活性测定作为一种在体内检测和定量淋巴细胞活化的方法的潜在实用性，已经进行了研究。开发了一种改良的酶测定法，以优化细胞或组织粗匀浆中活性的测量，从而便于处理多个样品。通过在测定混合物中加入双脱氧胸苷三磷酸（其他真核DNA聚合酶的抑制剂）来确保该测定对聚合酶α的特异性。在有丝分裂原刺激的体外系统中，发现DNA聚合酶α的活性与[3H]胸苷掺入密切相关。在3种不同的免疫介导或药物诱导的淋巴细胞反应的体内系统中，验证了聚合酶α方法用于定量淋巴细胞活化的实用性。
