15-deoxy-delta 12,14-prostaglandin J2. A prostaglandin D2 metabolite generated during inflammatory processes.
Prostaglandin D(2) (PGD(2)), a major cyclooxygenase product in a variety of tissues, readily undergoes dehydration to yield the cyclopentenone-type PGs of the J(2) series, such as 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ(2)), which have been suggested to exert anti-inflammatory effects in vivo. Meanwhile, the mechanism of these effects is not well understood and the natural site and the extent of its production in vivo remain unclear. In the present study, we raised a monoclonal antibody specific to 15d-PGJ(2) and determined its production in inflammation-related events. The monoclonal antibody (mAb11G2) was raised against the 15d-PGJ(2)-keyhole limpet hemocyanin conjugate and was found to recognize free 15d-PGJ(2) specifically. The presence of 15d-PGJ(2) in vivo was immunohistochemically verified in the cytoplasm of most of the foamy macrophages in human atherosclerotic plaques. In addition, the immunostaining of lipopolysaccharide-stimulated RAW264.7 macrophages with mAb11G2 demonstrated an enhanced intracellular accumulation of 15d-PGJ(2), suggesting that the PGD(2) metabolic pathway, generating the anti-inflammatory PGs, is indeed utilized in the cells during inflammation. The activation of macrophages also resulted in the extracellular production of PGD(2), which was associated with a significant increase in the extracellular 15d-PGJ(2) levels, and the extracellular 15d-PGJ(2) production was reproduced by incubating PGD(2) in a cell-free medium and in phosphate-buffered saline. Moreover, using a chiral high performance liquid chromatography method for separation of PGD(2) metabolites, we established a novel metabolic pathway, in which PGD(2) is converted to 15d-PGJ(2) via an albumin-independent mechanism.
Shibata
Takahiro
T
Laboratory of Food and Biodynamics, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan.
Kondo
Mitsuhiro
M
Osawa
Toshihiko
T
Shibata
Noriyuki
N
Kobayashi
Makio
M
Uchida
Koji
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001161:Arteriosclerosis; D002460:Cell Line; D002474:Cell-Free System; D002851:Chromatography, High Pressure Liquid; D051546:Cyclooxygenase 2; D003593:Cytoplasm / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007155:Immunologic Factors / Q000378:metabolism*; D007249:Inflammation; D007527:Isoenzymes / Q000378:metabolism; D008264:Macrophages / Q000276:immunology / Q000378:metabolism*; D008565:Membrane Proteins; D051379:Mice; D008954:Models, Biological; D008956:Models, Chemical; D010710:Phosphates / Q000378:metabolism; D015230:Prostaglandin D2 / Q000031:analogs & derivatives* / Q000378:metabolism*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D012709:Serum Albumin / Q000378:metabolism; D013997:Time Factors
C097240:15-deoxy-delta(12,14)-prostaglandin J2; D000911:Antibodies, Monoclonal; D007155:Immunologic Factors; D007527:Isoenzymes; D008565:Membrane Proteins; D010710:Phosphates; D012709:Serum Albumin; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D015230:Prostaglandin D2