A sensitive gas chromatography-mass spectrometry assay reveals increased levels of monohydroxyeicosatetraenoic acid isomers in human plasma after extracorporeal photoimmunotherapy and under in vitro ultraviolet A exposure.
Extracorporeal photoimmunotherapy (photopheresis) is a highly effective therapy in the treatment of various disorders. Although extracorporeal photoimmunotherapy has been successfully used for more than 10 y, its mechanism of action is still unclear. The formation of reactive oxygen species have been implicated in extracorporeal photoimmunotherapy, but malonyl dialdehyde as a marker of systemic lipid peroxidation did not increase significantly during treatment. To investigate further the involvement of reactive oxygen species in extracorporeal photoimmunotherapy, we have introduced a highly sensitive negative ion gas chromatography-mass spectrometry based method for quantitating oxygenated arachidonic acid isomers (hydroxyeicosatetraenoic acids) in plasma samples of patients treated with extracorporeal photoimmunotherapy. In the plasma of healthy volunteers pmole amounts of 2-, 3-, 5-, 8-12-, and 15-hydroxyeicosatetraenoic acid were detected and we observed a dose-dependent augmentation in these metabolites when the blood was irradiated with increasing doses of ultraviolet A in the presence of the photosensitizer 8-methoxypsoralen. Analysis of plasma samples obtained from patients before and after extracorporeal photoimmunotherapy revealed a characteristic increase in total hydroxyeicosatetraenoic acid levels, particularly of 5-hydroxyeicosatetraenoic acid which contributed 80% to the sum of all hydroxyeicosatetraenoic acid isomers. Chiral phase high-performance liquid chromatography indicated almost equal amounts of 5S- and 5R-hydroxyeicosatetraenoic acid suggesting that the majority of lipid peroxidation products are formed via nonenzymatic oxidation reactions.
Wiswedel
I
I
Department of Dermatology and Venereology, Institute of Clinical Chemistry and Pathological Biochemistry, Otto-von-Guericke-University, Magdeburg, Germany. ingrid.wiswedel@medizin.uni-magdeburg.de
Bohne
M
M
Hirsch
D
D
Kühn
H
H
Augustin
W
W
Gollnick
H
H
D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000097:blood*; D007167:Immunotherapy / Q000379:methods*; D007536:Isomerism; D016410:Lymphoma, T-Cell, Cutaneous / Q000628:therapy; D010778:Photochemotherapy / Q000379:methods; D012595:Scleroderma, Systemic / Q000628:therapy; D014466:Ultraviolet Rays
D006893:Hydroxyeicosatetraenoic Acids