Original Paper

Transgenic Research

, Volume 20, Issue 3, pp 625-641

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

  • M. MendicinoAffiliated withRevivicor, Inc.
  • , J. RamsoondarAffiliated withRevivicor, Inc.
  • , C. PhelpsAffiliated withRevivicor, Inc.
  • , T. VaughtAffiliated withRevivicor, Inc.
  • , S. BallAffiliated withRevivicor, Inc.
  • , T. LeRoithAffiliated withDepartment of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine
  • , J. MonahanAffiliated withRevivicor, Inc.
  • , S. ChenAffiliated withRevivicor, Inc.
  • , A. DandroAffiliated withRevivicor, Inc.
    • , J. BooneAffiliated withRevivicor, Inc.
    • , P. JobstAffiliated withRevivicor, Inc.
    • , A. VanceAffiliated withRevivicor, Inc.
    • , N. WertzAffiliated withDepartment of Microbiology and Interdisciplinary Immunology Program, University of Iowa
    • , Z. BergmanAffiliated withDepartment of Microbiology and Interdisciplinary Immunology Program, University of Iowa
    • , X-Z. SunAffiliated withDepartment of Microbiology and Interdisciplinary Immunology Program, University of Iowa
    • , I. PolejaevaAffiliated withRevivicor, Inc.
    • , J. ButlerAffiliated withDepartment of Microbiology and Interdisciplinary Immunology Program, University of Iowa
    • , Y. DaiAffiliated withRevivicor, Inc.
    • , D. AyaresAffiliated withRevivicor, Inc. Email author 
    • , K. WellsAffiliated withRevivicor, Inc.Division of Animal Sciences, University of Missouri Email author 

Abstract

A poly(A)-trap gene targeting strategy was used to disrupt the single functional heavy chain (HC) joining region (JH) of swine in primary fibroblasts. Genetically modified piglets were then generated via somatic cell nuclear transfer (SCNT) and bred to yield litters comprising JH wild-type littermate (+/+), JH heterozygous knockout (±) and JH 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 JH −/− pigs. Histochemical and immunohistochemical (IHC) studies failed to detect lymph node (LN) follicles or CD79α+ B cells, respectively in JH −/− pigs. T cell receptor (TCR)β transcription and T cells were detected in JH −/− pigs. When reared conventionally, JH −/− 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.

Keywords

Genetically engineered animal model Immunoglobulin heavy chain locus B cell and antibody deficiency Somatic cell nuclear transfer Humoral versus cellular immunity Human polyclonal antibody