Piccioli P, Ruberti F, Biocca S, Di Luzio A, Werge T M, Bradbury A, Cattaneo A
Consiglio Nazionale delle Ricerche, Istituto di Neurobiologia, Rome, Italy.
Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5611-5. doi: 10.1073/pnas.88.13.5611.
We present a strategy to study functional and/or developmental processes occurring in the nervous system, as well as in other systems, of mice. This strategy is based on the local expression of specific monoclonal antibodies (mAbs) by cells of the nervous system. As an application of this strategy, we report the cloning of the anti-substance P rat mAb NC1/34HL. Functional substance P-binding antibodies were reconstituted from the cloned variable domains by using vectors for expression in myeloma cells. With these and other vectors a general system for the cloning and expression of mAbs under a series of promoters (of the rat VGF8a gene, the neurofilament light-chain gene, and the methallothionein gene) has been created. The activity of these plasmids was confirmed by expressing the recombinant NC1/34HL mAb in GH3 pituitary cells, PC12 pheochromocytoma cells, and COS cells. DNA from the described constructs can be used to target the expression of the NC1/34HL mAb to the central nervous system of transgenic mice. This procedure will allow us to perturb substance P activity in a controlled way in order to dissect its multiple roles.
我们提出了一种策略，用于研究小鼠神经系统以及其他系统中发生的功能和/或发育过程。该策略基于神经系统细胞对特定单克隆抗体（mAb）的局部表达。作为此策略的应用，我们报告了抗P物质大鼠单克隆抗体NC1/34HL的克隆。通过使用在骨髓瘤细胞中表达的载体，从克隆的可变域中重建了具有功能的P物质结合抗体。利用这些载体和其他载体，创建了一个在一系列启动子（大鼠VGF8a基因、神经丝轻链基因和金属硫蛋白基因的启动子）控制下克隆和表达单克隆抗体的通用系统。通过在GH3垂体细胞、PC12嗜铬细胞瘤细胞和COS细胞中表达重组NC1/34HL单克隆抗体，证实了这些质粒的活性。来自所述构建体的DNA可用于将NC1/34HL单克隆抗体的表达靶向转基因小鼠的中枢神经系统。这一过程将使我们能够以可控方式干扰P物质的活性，以便剖析其多种作用。
