Mécheri S, Dannecker G, Dennig D, Poncet P, Hoffmann M K
Unité d'Immuno-Allergie, Institut Pasteur, Paris, France.
Mol Immunol. 1993 Apr;30(6):549-57. doi: 10.1016/0161-5890(93)90029-b.
In an attempt to induce an immune response against Mls-1a antigens by immunizing C57B1/6 mouse (Mls-1b) with purified B cells from DBA/2 mouse (Mls-1a), we generated a panel of monoclonal antibodies from which the 5B9.6 mAb, taken as a representative antibody, was thoroughly investigated. This antibody specifically reacts with B cells from all mouse strains studied including C57Bl/6 mice as shown by FACS analysis of double-antibody labelled spleen cells. Using enzyme immunoassays and immunoprecipitation techniques, 5B9.6 mAb was found to be specific for histones. Amino acid sequence analysis of a peptide derived from a 5B9.6-immunoprecipitated polypeptide from DBA/2 B cells showed a 100% homology with a sequence within H2B histones. Furthermore, 5B9.6 mAb was able to interact with the cell surface of 7OZ/3 cell line, known as a typical pre-B cell line. The presence of histones can be modulated on the surface of 7OZ/3 cells since this antigen was upregulated after exposure of these cells to a cocktail of IL-1 and cAMP. Finally, 5B9.6 mAb was shown to interact with freshly isolated B cells from human peripheral blood.
为了通过用来自DBA/2小鼠(Mls-1a)的纯化B细胞免疫C57B1/6小鼠(Mls-1b)来诱导针对Mls-1a抗原的免疫反应,我们产生了一组单克隆抗体,并对作为代表性抗体的5B9.6单克隆抗体进行了全面研究。如通过双抗体标记脾细胞的流式细胞术分析所示,该抗体与包括C57Bl/6小鼠在内的所有研究的小鼠品系的B细胞特异性反应。使用酶免疫测定和免疫沉淀技术,发现5B9.6单克隆抗体对组蛋白具有特异性。对从DBA/2 B细胞的5B9.6免疫沉淀多肽衍生的肽进行氨基酸序列分析,结果显示与H2B组蛋白内的序列具有100%的同源性。此外,5B9.6单克隆抗体能够与称为典型前B细胞系的7OZ/3细胞系的细胞表面相互作用。7OZ/3细胞表面的组蛋白存在可以被调节,因为这些细胞暴露于IL-1和cAMP的混合物后,这种抗原会上调。最后,5B9.6单克隆抗体显示与来自人外周血的新鲜分离的B细胞相互作用。