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Antibody Repertoire Development in Fetal and Neonatal Pigs. VII. Characterization of the Preimmune Light Chain Repertoire¹

John E. Butler^2,*, Nancy Wertz^*, Jishan Sun^*, Huang Wang^*, Patrick Chardon, Francois Piumi and Kevin Wells

^* Department of Microbiology and Interdisciplinary Immunology Program, University of Iowa, Iowa City, IA 52242; Institut National de la Recherche Agronomique, Institute of Molecular Biology, Jouy-en Josas, France; and Revivicor, Blacksburg, VA 24060

Combinatorial diversity is highly restricted in the preimmune porcine H chain repertoire compared with that in humans and mice. This raised the question of whether similar restriction characterized the preimmune L chain repertoire. In this study we present evidence that >90% of all expressed V genes in the porcine preimmune repertoire belong to three subfamilies of V genes that share 87% sequence similarity with human IGKV2. This porcine V family also shares sequence similarity with some, but not all, V genes from sheep. Hybridization with sperm DNA and sequence analyses of polynucleotides from overlapping bacterial artificial chromosome clones suggest swine possess 60 IGVK2 genes. The latter method also revealed that certain IGKV2 subfamilies are not expressed in the preimmune repertoire. Six members of an IGVK1 family were also expressed as part of the preimmune repertoire, and these shared 87% sequence similarity with human IGVK1. Five J segments, complete with recombination signal sequences and separated by 300 nt, were identified 3 kb upstream of a single C. Surprisingly, J2 accounted for >90% of all framework region 4 sequences in the preimmune repertoire. These findings show that swine use 10 IGVK2 genes from three of six subfamilies and preferentially one J segment to generate their preimmune repertoire. These studies, like those of porcine Ig constant regions and MHC genes, also indicate unexpected high sequence similarity with their human counterparts despite differences in phylogeny and the mechanism of repertoire diversification.