SIMULTANEOUS ESTABLISHMENT OF MONOCLONAL-ANTIBODIES SPECIFIC FOR EITHER CYCLOBUTANE PYRIMIDINE DIMER OR (6-4)PHOTOPRODUCT FROM THE SAME MOUSE IMMUNIZED WITH ULTRAVIOLET-IRRADIATED DNA
SIMULTANEOUS ESTABLISHMENT OF MONOCLONAL-ANTIBODIES SPECIFIC FOR EITHER CYCLOBUTANE PYRIMIDINE DIMER OR (6-4)PHOTOPRODUCT FROM THE SAME MOUSE IMMUNIZED WITH ULTRAVIOLET-IRRADIATED DNA
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DOI：
10.1111/j.1751-1097.1991.tb02010.x
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发表时间：
1991-08-01
影响因子：
3.3
通讯作者：
NIKAIDO, O
中科院分区：
文献类型：
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作者：
MORI, T;NAKANE, M;NIKAIDO, O
本文报道了六种新的抗紫外线（UV）损伤的单克隆抗体（TDM-2、TDM-3、64 M-2、64 M-3、64 M-4和64 M-5）。 在抗体表征实验中，发现两种TDM抗体显示与UV照射的DNA（UV-DNA）的剂量依赖性结合，在环丁烷嘧啶二聚体光复活后与UV-DNA的结合减少，与含有环丁烷胸腺嘧啶二聚体的DNA结合，并且在（6-4）光产物光异构化为杜瓦光产物后与UV-DNA的结合不变。 这些结果表明TDM单克隆抗体的表位是DNA中的环丁烷嘧啶二聚体。 另一方面，发现4种6 - 4 M抗体显示与UV-DNA的剂量依赖性结合，在环丁烷嘧啶二聚体光复活后与UV-DNA的结合不变，与含有胸腺嘧啶二聚体的DNA的结合不可检测，并且在（6-4）光产物的光异构化后与UV-DNA的结合减少。 这些结果表明，6 - 4 M抗体的表位是DNA中的（6-4）光产物。 这是第一次同时建立抗同一小鼠两种不同类型光损伤的单克隆抗体的报告。 通过使用这些单克隆抗体，我们已经成功地测量了环丁烷嘧啶二聚体和（6-4）光产物的DNA从人原代细胞照射生理紫外线剂量。
Six new monoclonal antibodies (TDM-2, TDM-3, 64M-2, 64M-3, 64M-4 and 64M-5) specific for ultraviolet (UV) induced DNA damage have been established. In the antibody characterization experiments, two TDM antibodies were found to show a dose-dependent binding to UV-irradiated DNA (UV-DNA), decrease of binding to UV-DNA after cyclobutane pyrimidine dimer photoreactivation, binding to DNA containing cyclobutane thymine dimers, and unchanged binding to UV-DNA after photoisomerization of (6-4)photoproducts to Dewar photoproducts. These results indicated that the epitope of TDM monoclonal antibodies was the cyclobutane pyrimidine dimer in DNA. On the other hand, four 64M antibodies were found to show a dose-dependent binding to UV-DNA, unchanged binding to UV-DNA after cyclobutane pyrimidine dimer photoreactivation, undetectable binding to DNA containing thymine dimers, and decrease of binding to UV-DNA after photoisomerization of (6-4)photoproducts. These results indicated that the epitope of 64M antibodies was the (6-4)photoproduct in DNA. This is the first report of the simultaneous establishment of monoclonal antibodies against the two different types of photolesions from the same mouse. By using these monoclonal antibodies, we have succeeded in measuring both cyclobutane pyrimidine dimers and (6-4)photoproducts in the DNA from human primary cells irradiated with physiological UV doses.