Ubiquitous LEA29Y Expression Blocks T Cell Co-Stimulation but Permits Sexual Reproduction in Genetically Modified Pigs

We have successfully established and characterized a genetically modified pig line with ubiquitous expression of LEA29Y, a human CTLA4-Ig derivate. LEA29Y binds human B7.1/CD80 and B7.2/CD86 with high affinity and is thus a potent inhibitor of T cell co-stimulation via this pathway. We have characterized the expression pattern and the biological function of the transgene as well as its impact on the porcine immune system and have evaluated the potential of these transgenic pigs to propagate via assisted breeding methods. The analysis of LEA29Y expression in serum and multiple organs of CAG-LEA transgenic pigs revealed that these animals produce a biologically active transgenic product at a considerable level. They present with an immune system affected by transgene expression, but can be maintained until sexual maturity and propagated by assisted reproduction techniques. Based on previous experience with pancreatic islets expressing LEA29Y, tissues from CAG-LEA29Y transgenic pigs should be protected against rejection by human T cells. Furthermore, their immune-compromised phenotype makes CAG-LEA29Y transgenic pigs an interesting large animal model for testing human cell therapies and will provide an important tool for further clarifying the LEA29Y mode of action

A review of the human vs. porcine female genital tract and associated immune system in the perspective of using minipigs as a model of human genital Chlamydia infection

International audienceAbstractSexually transmitted diseases constitute major health issues and their prevention and treatment continue to challenge the health care systems worldwide. Animal models are essential for a deeper understanding of the diseases and the development of safe and protective vaccines. Currently a good predictive non-rodent model is needed for the study of genital chlamydia in women. The pig has become an increasingly popular model for human diseases due to its close similarities to humans. The aim of this review is to compare the porcine and human female genital tract and associated immune system in the perspective of genital Chlamydia infection. The comparison of women and sows has shown that despite some gross anatomical differences, the structures and proportion of layers undergoing cyclic alterations are very similar. Reproductive hormonal cycles are closely related, only showing a slight difference in cycle length and source of luteolysing hormone. The epithelium and functional layers of the endometrium show similar cyclic changes. The immune system in pigs is very similar to that of humans, even though pigs have a higher percentage of CD4+/CD8+ double positive T cells. The genital immune system is also very similar in terms of the cyclic fluctuations in the mucosal antibody levels, but differs slightly regarding immune cell infiltration in the genital mucosa - predominantly due to the influx of neutrophils in the porcine endometrium during estrus. The vaginal flora in Göttingen Minipigs is not dominated by lactobacilli as in humans. The vaginal pH is around 7 in Göttingen Minipigs, compared to the more acidic vaginal pH around 3.5–5 in women. This review reveals important similarities between the human and porcine female reproductive tracts and proposes the pig as an advantageous supplementary model of human genital Chlamydia infection

Molecular cloning of porcine T cell receptor alpha, beta, gamma and delta chains using polymerase chain reaction fragments of the constant regions.

Fragments of the constant regions of porcine alpha, beta, delta and two types of gamma T cell receptor (TcR) chains were obtained by reverse transcription and polymerase chain reaction (PCR). Screening of a porcine peripheral T cell cDNA library with these PCR fragments led to the isolation of porcine TcR alpha, beta, gamma and delta chain clones. Sequence analysis of these clones and the respective PCR fragments demonstrated the existence of one alpha, one beta, three gamma and one delta chain isotype. Comparisons of the deduced amino acid sequences of the constant region with other species revealed a significant homology. For two of the three identified porcine gamma chain insertions of 38 and 40 amino acids were found within the hinge region. In addition, our sequence data demonstrate a high variability in the cytoplasmic C gamma domain among the three porcine TcR gamma isotypes as well as between species, which might be of structural and/or functional significance. Comparison with biochemical data indicate the existence of four porcine TcR gamma isotypes

Porcine T-cell receptors: molecular and biochemical characterization.

Two subclasses of CD3 associated T-cell receptors (TcR) have been described so far, consisting of either an alpha and beta chain (TcR alpha beta) or a gamma and delta chain (TcR gamma delta). Of the two subclasses, the TcR alpha beta is the one predominantly expressed on peripheral T lymphocytes of humans and rodents. TcR gamma delta T lymphocytes represent only a minor subset in these species. Among all mammalian species studied so far, swine showed the most diversified composition of the T-lymphocyte population characterized by the expression of CD4 and CD8 differentiation antigens. Besides CD4+CD8- and CD4-CD8+ T lymphocytes, CD4+CD8+ and CD4-CD8- T lymphocytes are prominent in the extrathymic T-lymphocyte compartment. Because of the lack of specific monoclonal antibodies (mAb), to date the porcine TcR can only be characterized with biochemical and molecular biological methods. TcR on porcine peripheral blood T lymphocytes with the phenotype CD4+ and/or CD8+ are characterized as 46-48 kDaR heterodimers which were supposed to represent the porcine TcR alpha beta. Biochemical analyses of the CD4-CD8- T lymphocytes revealed three distinct TcR gamma delta; all are characterized by a 40 kDa delta chain but differed in their gamma chains. One gamma chain with a molecular mass of 38 kDaR is preferentially expressed on CD4-CD8- T lymphocytes derived from peripheral blood; another chain with molecular mass of 37 kDaR is evenly distributed between CD4-CD8- T lymphocytes from blood and lymphoid tissues.(ABSTRACT TRUNCATED AT 250 WORDS