Buenafe A C, Makowski F F, Rittenberg M B
Department of Microbiology and Immunology, Oregon Health Sciences University, Portland 97201.
J Immunol. 1989 Jul 15;143(2):539-45.
We have identified four fine specificity groups reactive with the organophosphorus hapten Soman among 46 hybridomas generated in specific response to immunization with Soman-keyhole limpet hemocyanin (KLH). The different fine specificity groups do not appear to correlate with the use of particular V genes. Molecular analysis of VH genes demonstrates predominant use of VH J558 family members in hybridomas of all fine specificity groups although several different VH genes within this family as well as others are able to contribute. Diversity of VH gene usage was also apparent in primary IgM-producing hybridomas. In contrast, there appears to be restricted L chain usage; a large number (18/46) used the V kappa 1 family. Interestingly, the V kappa 1 family also plays an important role in the memory response to phosphocholine (PC)-KLH, a related organophosphate hapten which shares several structural features with Soman, particularly when coupled to protein carriers. The V kappa 1 C gene appears to predominate in the PC-KLH response. Restriction analysis of DNA from the V kappa 1-positive Soman-KLH-specific hybridomas suggests that a single V kappa 1 gene may be utilized by 17/18 but that this gene is different from V kappa 1 C and may be V kappa 1 A. We propose that members of the V kappa 1 family contribute favorably in generating combining sites that recognize all or part of the structural features shared by the two haptenic structures Soman and PC when they are coupled to protein. This most likely involves recognition of the phenyl linker moiety as the dominant feature. It appears that the L chain rather than the H chain may play a more significant role in forming the phenyl-Soman-specific combining site and perhaps the combining sites for phenyl or ring structures in general.
在针对梭曼-钥孔戚血蓝蛋白（KLH）免疫产生特异性反应而生成的46个杂交瘤中，我们鉴定出了4个与有机磷半抗原梭曼反应的精细特异性组。不同的精细特异性组似乎与特定V基因的使用无关。VH基因的分子分析表明，在所有精细特异性组的杂交瘤中，VH J558家族成员占主导地位，尽管该家族内的几个不同VH基因以及其他基因也有贡献。VH基因使用的多样性在初级产生IgM的杂交瘤中也很明显。相比之下，轻链的使用似乎受到限制；大量（18/46）使用了Vκ1家族。有趣的是，Vκ1家族在对磷胆碱（PC）-KLH的记忆反应中也起重要作用，PC-KLH是一种相关的有机磷酸半抗原，与梭曼有几个共同的结构特征，特别是与蛋白质载体偶联时。Vκ1 C基因似乎在PC-KLH反应中占主导地位。对Vκ1阳性的梭曼-KLH特异性杂交瘤的DNA进行限制性分析表明，17/18个杂交瘤可能使用单个Vκ1基因，但该基因与Vκ1 C不同，可能是Vκ1 A。我们提出，当Vκ1家族成员与蛋白质偶联时，它们在产生识别梭曼和PC这两种半抗原结构的全部或部分共同结构特征的结合位点方面发挥了有利作用。这很可能涉及将苯基连接基团识别为主要特征。似乎轻链而非重链在形成苯基-梭曼特异性结合位点以及可能一般的苯基或环结构结合位点方面可能发挥更重要的作用。
