Production and characterization of transgenic pigs expressing porcine CTLA4-Ig.

Background: Inhibition of the T-cell-mediated immune response is a necessary component of preventing rejection following xenotransplantation with pig alpha1,3-galactosyltransferase gene-knockout (GTKO) organs. Cytotoxic T lymphocyte-associated antigen (CTLA4) is a co-stimulatory molecule that inhibits T-cell activity and may be useful in prolonging graft rejection.

Methods: An expression vector was built containing the extracellular coding region of porcine (p) CTLA4 fused to the hinge and CH2/CH3 regions of human IgG1 (pCTLA4-Ig).

Production of alpha 1,3-galactosyltransferase-knockout cloned pigs expressing human alpha 1,2-fucosylosyltransferase.

The production of genetically engineered pigs as xenotransplant donors aims to solve the severe shortage of organs for transplantation in humans. The first barrier to successful xenotransplantation is hyperacute rejection (HAR). HAR is a rapid and massive humoral immune response directed against the pig carbohydrate Galalpha 1,3-Gal epitope, which is synthesized by alpha 1,3-galactosyltransferase (alpha1,3-GT).

Na(+)/Ca(2+) exchanger in porcine oocytes.

The presence of the Na(+)/Ca(2+) exchange mechanism was investigated in porcine oocytes. Immature and in vitro-matured oocytes were loaded with the Ca(2+)-sensitive fluorescent dye fura 2 and changes in the intracellular free Ca(2+) concentration ([Ca(2+)](i)) were monitored after altering the Na(+) concentration gradient across the plasma membrane. Decreasing the extracellular Na(+) concentration induced an increase in [Ca(2+)](i) possibly by a Ca(2+) influx via the Na(+)/Ca(2+) exchanger.

The use of nuclear transfer to produce transgenic pigs.

Manipulation of the pig genome has the potential to improve pig production and offers powerful biomedical applications. Genetic manipulation of mammals has been possible for over two decades, but the technology available has proven both difficult and inefficient. The development of new techniques to enhance efficiency and overcome the complications of random insertion is of importance.

Capacitative calcium entry mechanism in porcine oocytes.

The presence of the capacitative Ca(2+) entry mechanism was investigated in porcine oocytes. In vitro-matured oocytes were treated with thapsigargin in Ca(2+)-free medium for 3 h to deplete intracellular calcium stores. After restoring extracellular calcium, a large calcium influx was measured by using the calcium indicator dye fura-2, indicating capacitative Ca(2+) entry.