Le Bihan, Sylvie, Recombinant Antibody Technology Ltd. [Cambridge, UK], Babraham Bioscience Technologies Ltd [Cambridge, UK], Centre de Recherche en Transplantation et Immunologie (U1064 Inserm - CRTI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), Open Monoclonal Technology, Inc., Ligand Pharmaceuticals, OMT., Biogenouest., Région Pays de la Loire, France.
المصدر:
Journal of Immunological Methods Journal of Immunological Methods, 2013, 400-401, pp.78-86. ⟨10.1016/j.jim.2013.10.007⟩
International audience; Expression of human antibody repertoires in transgenic animals has been accomplished by introducing large human Ig loci into mice and, more recently, a chimeric IgH locus into rats. With human V[H] , D and J[H] genes linked to the rat C-region antibody expression was significantly increased, similar to wild-type levels not found with fully human constructs. Here we compare four rat-lines containing the same human V[H]-region (comprising 22 V[H]s, all Ds and all J[H]s in natural configuration) but linked to different rat C[H]-genes and regulatory sequences. The endogenous IgH locus was silenced by zinc-finger nucleases. After breeding, all lines produced exclusively chimeric human H-chain with near normal IgM levels. However, in two lines poor IgG expression and inefficient immune responses were observed, implying that high expression, class-switching and hypermutation are linked to optimal enhancer function provided by the large regulatory region at the 3′ end of the IgH locus. Furthermore, exclusion of Cδ and its downstream interval region may assist recombination. Highly diverse IgG and immune responses similar to normal rats were identified in two strains carrying diverse and differently spaced C-genes.
