Wagner S D, Williams G T, Larson T, Neuberger M S, Kitamura D, Rajewsky K, Xian J, Brüggemann M
Medical Research Council Laboratory of Molecular Biology, Cambridge, UK.
Nucleic Acids Res. 1994 Apr 25;22(8):1389-93. doi: 10.1093/nar/22.8.1389.
One approach to the production of human monoclonal antibodies focusses on the creation of transgenic mice bearing human immunoglobulin gene miniloci. Whilst such loci undergo lymphoid-specific gene rearrangement, only a small proportion of mouse B cells express the human immunoglobulin chains; the miniloci thus contribute poorly to serum immunoglobulin. Attributing this poor performance to competition between the transgenic and endogenous immunoglobulin loci, we crossed mice bearing a human immunoglobulin heavy-chain (HulgH) minilocus with animals that had been rendered B cell-deficient by disruption of their endogenous heavy-chain locus. The results were dramatic: the human minilocus rescued B cell differentiation such that effectively all B cells now expressed human mu chains. The concentration of antibody in the mouse serum recognised by anti-human mu increased to a concentration about one sixth that in human serum. The HulgH antibodies are heterogenous with diversity being generated by both combinatorial and junctional processes. Following antigen challenge, specific antibody is elicited but at low titre.
生产人单克隆抗体的一种方法聚焦于创建携带人免疫球蛋白基因微基因座的转基因小鼠。虽然这些基因座会发生淋巴细胞特异性基因重排，但只有一小部分小鼠B细胞表达人免疫球蛋白链；因此，微基因座对血清免疫球蛋白的贡献很小。将这种不佳表现归因于转基因和内源性免疫球蛋白基因座之间的竞争，我们将携带人免疫球蛋白重链（HulgH）微基因座的小鼠与通过破坏其内源性重链基因座而导致B细胞缺陷的动物进行杂交。结果令人瞩目：人微基因座挽救了B细胞分化，使得实际上所有B细胞现在都表达人μ链。抗人μ识别的小鼠血清中抗体浓度增加到约为人血清中浓度的六分之一。HulgH抗体是异质的，其多样性由组合和连接过程产生。抗原刺激后，会产生特异性抗体，但滴度较低。
