Monoclonal antibodies to DNA and RNA from NZB/NZW F1 mice: antigenic specificities and NH2 terminal amino acid sequences.
Monoclonal antibodies to DNA and RNA from NZB/NZW F1 mice: antigenic specificities and NH2 terminal amino acid sequences.
复制标题
针对 NZB/NZW F1 小鼠 DNA 和 RNA 的单克隆抗体：抗原特异性和 NH2 末端氨基酸序列。
DOI：
10.4049/jimmunol.133.1.489
复制
发表时间：
1984
影响因子：
4.4
通讯作者：
S. Rudikoff
中科院分区：
文献类型：
--
作者：
D. Eilat;M. Hochberg;J. Pumphrey;S. Rudikoff
将未免疫的NZB/NZW F1雌性小鼠脾细胞与BALB/c骨髓瘤细胞融合，制备了两株抗DNA杂交瘤自身抗体（A52、D42）。将单克隆抗体纯化至同质，并分析其抗原结合特异性。这两种抗DNA抗体结合单链、双链和超螺旋DNA，对单链构象具有明显的偏好。用合成的多核苷酸进行的竞争实验以及链重建实验表明，核酸的糖-磷酸骨架和杂环碱基对于抗体识别是必不可少的。A52和两个RNA结合杂交瘤蛋白的氨基末端序列分析显示，来自所有三个的重链是VHII亚组的成员，并且A52轻链与VK8亚组同源。发现D42重链类似于VHIII亚组的磷酸胆碱结合杂交瘤。
Two anti-DNA hybridoma autoantibodies ( A52 , D42 ) were prepared by fusing spleen cells from unimmunized NZB/NZW F1 female mice with BALB/c myeloma cells. The monoclonal antibodies were purified to homogeneity and were analyzed for their antigen-binding specificities. The two anti-DNA antibodies bound single-stranded, double-stranded, and supercoiled DNA, with a marked preference for the single-stranded conformation. Competition experiments performed with synthetic polynucleotides, as well as chain reconstitution experiments, indicated that both the sugar-phosphate backbone and the heterocyclic bases of the nucleic acid are essential for antibody recognition. Amino terminal sequence analysis of A52 and two RNA-binding hybridoma proteins revealed that the heavy chains from all three were members of the VHII subgroup and that the A52 light chain was homologous to the VK8 subgroup. The D42 heavy chain was found to be similar to a phosphocholine-binding hybridoma of the VHIII subgroup.