The major source of endogenous prostaglandin D2 production is likely antigen-presenting cells. Localization of glutathione-requiring prostaglandin D synthetase in histiocytes, dendritic, and Kupffer cells in various rat tissues.
The cellular localization of glutathione-requiring PGD synthetase, which catalyzes the predominant formation of PGD2 in various peripheral tissues, was investigated in adult rats by immunoperoxidase-staining with a polyclonal antibody specific for this enzyme. Although the 25 N-terminal amino acid residues of synthetase are 56% identical and 76% similar to those of several rat glutathione S-transferase subunits, the antibody cross-reacted only with synthetase in dot blotting and was nearly completely inactive with all transferase isozymes thus far purified. In Western blotting after SDS-PAGE of crude extracts of rat spleen, the antibody showed a single positive band at the same position as that of the purified enzyme (Mr = 26,000). The positive immunocytochemical stain was found in a number of histiocytes and/or dendritic cells in spleen, thymus, and Peyer's patch of intestine. The immunostain was also observed in such cells in lamina propria of the villus in small intestine and colon, in submucosal layer of stomach, and in Kupffer cells in liver. Immunoelectron microscopy confirmed that immunoreactivity of this enzyme was distributed in cytoplasm of those cells. Such immunoreactive cells were not observed in brain, spinal cord, kidney, heart, testis, and skeletal muscle. These observations suggest that PGD2 is produced by glutathione-requiring PGD synthetase localized in these types of APC in various tissues and may play a critical role in dictating the progression of immune responses.
Urade
Y
Y
Department of Enzymes and Metabolism, Osaka Bioscience Institute, Japan.
Ujihara
M
M
Horiguchi
Y
Y
Ikai
K
K
Hayaishi
O
O
D000595:Amino Acid Sequence; D000818:Animals; D000938:Antigen-Presenting Cells / Q000378:metabolism*; D015153:Blotting, Western; D003713:Dendritic Cells / Q000201:enzymology*; D005978:Glutathione / Q000378:metabolism; D006644:Histiocytes / Q000201:enzymology*; D007124:Immunoenzyme Techniques; D007421:Intestine, Small / Q000201:enzymology; D007728:Kupffer Cells / Q000201:enzymology*; D008969:Molecular Sequence Data; D015230:Prostaglandin D2 / Q000096:biosynthesis*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism*; D051381:Rats; D013154:Spleen / Q000201:enzymology; D013950:Thymus Gland / Q000201:enzymology
D011451:Prostaglandin-Endoperoxide Synthases; D005978:Glutathione; D015230:Prostaglandin D2