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1.
Clin Vaccine Immunol ; 20(9): 1466-72, 2013 Sep.
Article in English | MEDLINE | ID: mdl-23885030

ABSTRACT

Haemophilus parasuis causes Glässer's disease, a syndrome of polyserositis, meningitis, and arthritis in swine. Previous studies with H. parasuis have revealed virulence disparity among isolates and inconsistent heterologous protection. In this study, virulence, direct transmission, and heterologous protection of 4 isolates of H. parasuis (SW114, 12939, MN-H, and 29755) were evaluated using a highly susceptible pig model. In an initial experiment, isolates 12939, MN-H, and 29755 caused Glässer's disease, while strain SW114 failed to cause any clinical signs of disease. One pig from each group challenged with MN-H or 29755 failed to develop clinical disease but was able to transmit H. parasuis to noninfected pigs, which subsequently developed Glässer's disease. Pigs colonized with SW114, 29755, or MN-H that were free of clinical disease were protected from a subsequent challenge with isolate 12939. In a following experiment, pigs vaccinated with strain SW114 given as either a bacterin intramuscularly or a live intranasal vaccine were protected from subsequent challenge with isolate 12939; however, some pigs given live SW114 developed arthritis. Overall these studies demonstrated that pigs infected with virulent isolates of H. parasuis can remain healthy and serve as reservoirs for transmission to naive pigs and that heterologous protection among H. parasuis isolates is possible. In addition, further attenuation of strain SW114 is necessary if it is to be used as a live vaccine.


Subject(s)
Cross Protection , Haemophilus Infections/immunology , Haemophilus Infections/prevention & control , Haemophilus parasuis/immunology , Haemophilus parasuis/pathogenicity , Swine Diseases/immunology , Swine Diseases/prevention & control , Animals , Disease Models, Animal , Haemophilus Infections/transmission , Haemophilus Vaccines/administration & dosage , Haemophilus Vaccines/immunology , Haemophilus parasuis/isolation & purification , Swine , Swine Diseases/transmission
2.
Cloning Stem Cells ; 5(2): 117-21, 2003.
Article in English | MEDLINE | ID: mdl-12930623

ABSTRACT

Nuclear transfer technology allows for the reprogramming of somatic cells, and the production of embryonic stem cells and animals that are genetically identical in terms of nuclear DNA to the parental somatic cell. It is assumed that these products of nuclear transfer technology will be immunologically compatible to each other in spite of the fact that there are data that show differences in the expression patterns and phenotypes between animals produced by nuclear transfer. We have produced a series of cloned pigs from embryonic fibroblasts. Microsatellite analysis was used to confirm that the clones were genetically identical. Skin transplants were performed to assess immunological reactivity. Skin transplants between genetically identical cloned pigs were accepted, whereas third party grafts were rejected. Histological analysis of the grafts showed edema and mononuclear cell infiltrates in the recipient's skin in rejected grafts and not in grafts that were accepted. Our data supports the notion that genetically identical cloned pigs are immunologically compatible.


Subject(s)
Cloning, Organism , Skin Transplantation/immunology , Swine , Animals , Cell Line , Embryo, Mammalian , Fibroblasts , Graft Survival , Nuclear Transfer Techniques
3.
Transplantation ; 75(4): 430-6, 2003 Feb 27.
Article in English | MEDLINE | ID: mdl-12605105

ABSTRACT

BACKGROUND: The major antigen recognized on pig tissue by primate antibodies is a terminal galalpha1-3gal carbohydrate structure (gal antigen) present on glycolipids and glycoproteins. The production of animals from somatic cells allows for the inactivation of specific genes. It is anticipated that the complete inactivation of the gene encoding alpha1-3 galactosyltransferase, the enzyme that synthesizes the galalpha1-3gal linkage, will result in loss of that antigen from pig organs and tissue and will provide a survival benefit in pig-to-primate xenotransplants. METHODS: Positive-negative selection was used to produce fetal-pig fibroblasts that were a heterozygous knockout (+/-) of the alpha1-3 galactosyltransferase gene. Nuclear transfer of these cells generated pig embryos and live born pigs with the appropriate genotype. Using a novel selection method with cells from (+/-) embryos, we produced homozygous (-/-) fetal-pig fibroblast cells. RESULTS: Southern blot analysis of the alpha1-3 galactosyltransferase gene showed that we had produced (+/-) pig embryos, (+/-) live born pigs, and (-/-) pig-fetal fibroblast cells. Fluorescence-activated cell sorter (FACS) analysis with some, but not all, mouse anti-gal monoclonal antibodies and sensitized human serum showed that (-/-) cells still synthesized the gal antigen at 1 to 2% of the level of control heterozygous cells. CONCLUSIONS: Fetal-pig fibroblasts homozygous for the knockout of the alpha1-3 galactosyltransferase gene appear to express low but detectable levels of the gal antigen.


Subject(s)
Disaccharides/immunology , Disaccharides/metabolism , Fetal Tissue Transplantation/immunology , Fibroblasts/immunology , Galactosyltransferases/genetics , Animals , Antibodies, Monoclonal/immunology , Cells, Cultured , Epitopes , Fibroblasts/cytology , Flow Cytometry , Heterozygote , Swine
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