Cloning of pig serine proteinase inhibitor 9 and its use in protecting against apoptosis.
The activity of granzyme B, a main effector molecule of natural killer (NK) cells and cytotoxic T lymphocytes, is regulated by the intracellular serine proteinase inhibitor 9 (PI-9). Pig PI-9 was first cloned, and the sequences that encode pig PI-9, including the start codon and stop codon, were identified. The cDNA was inserted into the cloning site of pCXN2 (chicken beta actin promoter and cytomegalovirus enhancer), transfected into pig endothelial cells (PEC), and several stable PEC clones were established. An NK cell-mediated cytolysis test was next applied to the PEC clones, using YT cells (an NK-like cell line). The PEC transfectants with pig PI-9 had a significant inhibitory effect on NK cell-mediated PEC lysis. The overexpression of the anti-apoptotic molecule, pig PI-9, has the potential for use in protecting graft cells from human NK cells.
Matsunami
Katsuyoshi
K
Division of Organ Transplantation, Department of Molecular Therapeutics, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Kondo
Akihiro
A
Nakatsu
Shino
S
Omori
Takeshi
T
Nakagawa
Takatoshi
T
Otsuka
Hideaki
H
Fukuzawa
Masahiro
M
Miyagawa
Shuji
S
D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D042783:Endothelial Cells / Q000502:physiology*; D006084:Graft Rejection / Q000503:physiopathology*; D006801:Humans; D007694:Killer Cells, Natural / Q000502:physiology*; D007770:L-Lactate Dehydrogenase; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015842:Serine Proteinase Inhibitors / Q000235:genetics / Q000502:physiology*; D013552:Swine; D014162:Transfection
D012333:RNA, Messenger; D015842:Serine Proteinase Inhibitors; D007770:L-Lactate Dehydrogenase