Generation of recombination activating gene-1-deficient neonatal piglets: a model of T and B cell deficient severe combined immune deficiency.
Generation of recombination activating gene-1-deficient neonatal piglets: a model of T and B cell deficient severe combined immune deficiency.
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DOI：
10.1371/journal.pone.0113833
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Nakauchi H
中科院分区：
文献类型：
--
作者：
Ito T;Sendai Y;Yamazaki S;Seki-Soma M;Hirose K;Watanabe M;Fukawa K;Nakauchi H
严重联合免疫缺陷（SCID）是一种非常重要的小鼠研究模型，并且SCID小鼠被广泛应用，但关于SCID猪模型的报道却很少。因此，需要在这一领域进行进一步的研究。在本研究中，我们采用体细胞核移植（SCNT）技术，利用荧光活化细胞分选（FACS）和组织学技术对重组激活基因-1 （Rag-1）缺陷的新生仔猪进行了基因靶向和分析。我们构建了猪Rag-1基因外显子2区的靶向载体，并利用SCNT获得了Rag-1基因杂合/纯合敲除细胞菌落。我们培育了两只缺乏拉格-1的新生仔猪，并与野生型新生仔猪进行了比较。FACS分析显示，Rag-1的破坏导致外周血单核细胞中免疫球蛋白m阳性B细胞和cd3阳性T细胞的缺乏。与FACS分析一致，组织学分析显示rag -1缺陷仔猪的脾脏、肠系膜淋巴结（MLNs）和胸腺中存在结构缺陷和成熟淋巴细胞缺失。这些结果证实了Rag-1对于猪淋巴细胞的产生是必需的，并且Rag-1缺陷仔猪表现出与啮齿动物和人类相似的T和B细胞缺陷SCID （T-B-SCID）表型。本研究中产生的rag1缺乏症T-B-SCID猪可以作为实验室、转化和生物医学研究的合适的通用模型，包括人源化模型的开发和多能干细胞的评估。
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.
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影响因子：
3.7
作者：
Akkina R
通讯作者：
Akkina R
影响因子：
64.5
作者：
MCBLANE, JF;VANGENT, DC;OETTINGER, MA
通讯作者：
OETTINGER, MA
具有严重联合免疫缺陷的 RAG1/2 基因敲除猪
影响因子：
4.4
作者：
Huang, Jiao;Guo, Xiaogang;Lai, Liangxue
通讯作者：
Lai, Liangxue
影响因子：
2.5
作者：
Hwang, Kyu-Chan;Cho, Scong-Keun;Kim, Jin-Hoi
通讯作者：
Kim, Jin-Hoi
影响因子：
20.3
作者：
Kollet, O;Peled, A;Lapidot, T
通讯作者：
Lapidot, T