NDLI: Generation of antibody- and B cell-deficient pigs by targeted disruption of the J-region gene segment of the heavy chain locus

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Generation of antibody- and B cell-deficient pigs by targeted disruption of the J-region gene segment of the heavy chain locus

Content Provider	Springer Nature Link
Author	Mendici, M. Ramsoondar, J. Phelps, C. Vaught, T. Ball, S. LeRoith, T. Monahan, J. Chen, S. Dandro, A. Boone, J. Jobst, P. Vance, A. Wertz, N. Bergman, Z. Sun, X Z. Polejaeva, I. Butler, J. Dai, Y. Ayares, D. Wells, K.
Copyright Year	2010
Abstract	A poly(A)-trap gene targeting strategy was used to disrupt the single functional heavy chain (HC) joining region (J$_{H}$) of swine in primary fibroblasts. Genetically modified piglets were then generated via somatic cell nuclear transfer (SCNT) and bred to yield litters comprising J$_{H}$ wild-type littermate (+/+), J$_{H}$ heterozygous knockout (±) and J$_{H}$ homozygous knockout (−/−) piglets in the expected Mendelian ratio of 1:2:1. There are only two other targeted loci previously published in swine, and this is the first successful poly(A)-trap strategy ever published in a livestock species. In either blood or secondary lymphoid tissues, flow cytometry, RT-PCR and ELISA detected no circulating IgM$^{+}$ B cells, and no transcription or secretion of immunoglobulin (Ig) isotypes, respectively in J$_{H}$ −/− pigs. Histochemical and immunohistochemical (IHC) studies failed to detect lymph node (LN) follicles or CD79α$^{+}$ B cells, respectively in J$_{H}$ −/− pigs. T cell receptor (TCR)$_{β}$ transcription and T cells were detected in J$_{H}$ −/− pigs. When reared conventionally, J$_{H}$ −/− pigs succumbed to bacterial infections after weaning. These antibody (Ab)- and B cell-deficient pigs have significant value as models for both veterinary and human research to discriminate cellular and humoral protective immunity to infectious agents. Thus, these pigs may aid in vaccine development for infectious agents such as the pandemic porcine reproductive and respiratory syndrome virus (PRRSV) and H1N1 swine flu. These pigs are also a first significant step towards generating a pig that expresses fully human, antigen-specific polyclonal Ab to target numerous incurable infectious diseases with high unmet clinical need.
Starting Page	625
Ending Page	641
Page Count	17
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	20
Issue Number	3
e-ISSN	15739368
Language	English
Publisher	Springer Netherlands
Publisher Date	2010-09-26
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Genetically engineered animal model Immunoglobulin heavy chain locus B cell and antibody deficiency Somatic cell nuclear transfer Humoral versus cellular immunity Human polyclonal antibody Animal Genetics and Genomics Plant Sciences Molecular Medicine Plant Genetics & Genomics Human Genetics
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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