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

Autor: A. Vance, Todd Vaught, Jeff A. Monahan, Irina A. Polejaeva, Peter M. Jobst, Y. Dai, Jeremy Boone, M. Mendicino, Carol Phelps, Kevin D. Wells, David Ayares, LeRoith T, J. Ramsoondar, Xiu-Zhu Sun, Dandro A, Suyapa Ball, John E. Butler, Nancy Wertz, Bergman Z, S. Chen
Rok vydání: 2010
Předmět:
Swine
Somatic cell
T-Lymphocytes
Transfection
Immunoglobulin E
Antibodies
Flow cytometry
Human polyclonal antibody
Genetics
medicine
Animals
Humans
B cell and antibody deficiency
Cells
Cultured
B cell
Somatic cell nuclear transfer
Original Paper
B-Lymphocytes
biology
medicine.diagnostic_test
Humoral versus cellular immunity
T-cell receptor
Bacterial Infections
Fibroblasts
Immunoglobulin heavy chain locus
Porcine reproductive and respiratory syndrome virus
biology.organism_classification
Immunohistochemistry
Molecular biology
Immunoglobulin Isotypes
Disease Models
Animal
medicine.anatomical_structure
Animals
Newborn
Polyclonal antibodies
Gene Targeting
biology.protein
Genetically engineered animal model
Animal Science and Zoology
Antibody
Genetic Engineering
Immunoglobulin Heavy Chains
Poly A
Agronomy and Crop Science
Biotechnology
Zdroj: Transgenic Research
ISSN: 1573-9368
0962-8819
DOI: 10.1007/s11248-010-9444-z
Popis: 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.
Databáze: OpenAIRE
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