Matute C, Tigyi G J, Miledi R
Department of Psychobiology, University of California, Irvine 92717.
J Neurosci Res. 1991 May;29(1):77-86. doi: 10.1002/jnr.490290109.
A novel approach was developed to raise a panel of monoclonal antibodies (mAb) against brain antigens using Xenopus oocytes as immunological vectors. Xenopus oocytes were injected to express proteins encoded by brain-derived mRNA extracted from rat cerebral cortex. A crude membrane preparation from mRNA-injected oocytes was then used to immunize mice previously rendered immunotolerant to native oocyte membranes. mAb reacting with cryostat cut sections from rat brain were selected and further characterized by immunohistological and immunobiochemical techniques. Several mAb recognized brain specific antigens, including some that were cell type specific and others that revealed a regional binding pattern. A particular group of antibodies recognized an epitope localized exclusively to the cerebellar pinceau terminals. Although some of the hybridomas found in this panel may be products of natural autoreactive lymphocytes, the presence of a specific immune response to mRNA expression products is discussed. These results indicate that mRNA injected oocytes are useful tools to raise mAb to study the molecular diversity of the nervous system.
开发了一种新方法，利用非洲爪蟾卵母细胞作为免疫载体，制备针对脑抗原的一组单克隆抗体（mAb）。将非洲爪蟾卵母细胞注射以表达从大鼠大脑皮层提取的脑源性mRNA编码的蛋白质。然后，用注射了mRNA的卵母细胞的粗制膜制剂免疫先前对天然卵母细胞膜产生免疫耐受的小鼠。选择与大鼠脑冰冻切片反应的单克隆抗体，并通过免疫组织学和免疫生物化学技术进一步表征。几种单克隆抗体识别脑特异性抗原，包括一些细胞类型特异性抗原和其他显示区域结合模式的抗原。一组特定的抗体识别仅定位于小脑夹部终末的表位。虽然该组中发现的一些杂交瘤可能是天然自身反应性淋巴细胞的产物，但本文讨论了对mRNA表达产物存在特异性免疫反应的情况。这些结果表明，注射了mRNA的卵母细胞是制备单克隆抗体以研究神经系统分子多样性的有用工具。
