title: Generation of recombination activating gene-1-deficient neonatal piglets: a model of T and B cell deficient severe combined immune deficiency.
abstract: Although severe combined immune deficiency (SCID) is a very important research model for mice and SCID mice are widely used, there are only few reports describing the SCID pig models. Therefore, additional research in this area is needed. In this study, we describe the generation of Recombination activating gene-1 (Rag-1)-deficient neonatal piglets in Duroc breed using somatic cell nuclear transfer (SCNT) with gene targeting and analysis using fluorescence-activated cell sorting (FACS) and histology. We constructed porcine Rag-1 gene targeting vectors for the Exon 2 region and obtained heterozygous/homozygous Rag-1 knockout cell colonies using SCNT. We generated two Rag-1-deficient neonatal piglets and compared them with wild-type neonatal piglets. FACS analysis showed that Rag-1 disruption causes a lack of Immunoglobulin M-positive B cells and CD3-positive T cells in peripheral blood mononuclear cells. Consistent with FACS analysis, histological analysis revealed structural defects and an absence of mature lymphocytes in the spleen, mesenteric lymph node (MLNs), and thymus in Rag-1-deficient piglets. These results confirm that Rag-1 is necessary for the generation of lymphocytes in pigs, and Rag-1-deficient piglets exhibit a T and B cell deficient SCID (T-B-SCID) phenotype similar to that of rodents and humans. The T-B-SCID pigs with Rag-1 deficiency generated in this study could be a suitably versatile model for laboratory, translational, and biomedical research, including the development of a humanized model and assessment of pluripotent stem cells.
authors.affiliation: Pig Breeding Laboratory, Central Research Institute for Feed and Livestock, ZEN-NOH (National Federation of Agricultural Co-operative Associations), Hokkaido, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Ito
authors.affiliation: Biological Sciences Section, Central Research Institute for Feed and Livestock, ZEN-NOH (National Federation of Agricultural Co-operative Associations), Ibaraki, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Sendai
authors.affiliation: Division of Stem Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan; Japan Science and Technology Agency, Exploratory Research for Advanced Technology (ERATO), Nakauchi Stem Cell and Organ Regeneration Project, Tokyo, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yamazaki
authors.affiliation: Zen-noh Institute of Animal Health, ZEN-NOH (National Federation of Agricultural Co-operative Associations), Chiba, Japan.
authors.forename: Marie
authors.initials: M
authors.lastname: Seki-Soma
authors.affiliation: Pig Breeding Laboratory, Central Research Institute for Feed and Livestock, ZEN-NOH (National Federation of Agricultural Co-operative Associations), Hokkaido, Japan.
authors.forename: Kensuke
authors.initials: K
authors.lastname: Hirose
authors.affiliation: Division of Stem Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan; Japan Science and Technology Agency, Exploratory Research for Advanced Technology (ERATO), Nakauchi Stem Cell and Organ Regeneration Project, Tokyo, Japan.
authors.forename: Motoo
authors.initials: M
authors.lastname: Watanabe
authors.affiliation: Pig Breeding Laboratory, Central Research Institute for Feed and Livestock, ZEN-NOH (National Federation of Agricultural Co-operative Associations), Hokkaido, Japan.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Fukawa
authors.affiliation: Division of Stem Cell Therapy, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan; Japan Science and Technology Agency, Exploratory Research for Advanced Technology (ERATO), Nakauchi Stem Cell and Organ Regeneration Project, Tokyo, Japan.
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Nakauchi
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001402:B-Lymphocytes / Q000378:metabolism*; D002478:Cells, Cultured; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000166:cytology; D055786:Gene Knockout Techniques; D018398:Homeodomain Proteins / Q000235:genetics*; D053652:Nuclear Transfer Techniques; D016511:Severe Combined Immunodeficiency / Q000235:genetics* / Q000276:immunology / Q000473:pathology; D013552:Swine; D013601:T-Lymphocytes / Q000378:metabolism*
chemical_list: D018398:Homeodomain Proteins; C064658:RAG-1 protein