Xenopus oocytes as immunological vectors to produce monoclonal antibodies to rat brain antigens.
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.
Matute
C
C
Department of Psychobiology, University of California, Irvine 92717.
Tigyi
G J
GJ
Miledi
R
R
D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000276:immunology*; D001921:Brain / Q000276:immunology*; D002459:Cell Fusion; D003001:Cloning, Molecular; D003429:Cross Reactions; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009865:Oocytes / Q000276:immunology*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000378:metabolism; D014981:Xenopus
D000911:Antibodies, Monoclonal; D000941:Antigens; D012333:RNA, Messenger