Abstract
A method was established to measure tryptophan and kynurenine in serum simultaneously. Tryptophan is converted to kynurenine by the action of the enzyme indoleamine 2,3-dioxygenase induced by interferon-γ (IFN-γ). Since IFN-γ is a Th1-cell derived cytokine, an increased tryptophan degradation rate via the kynurenine pathway can be found when the cellular immune system is activated as it is, e.g., in viral infections or in autoimmune diseases. Thus, the ratio kynurenine per tryptophan provides a possibility to estimate IFN-γ activity in vivo and furthermore reflects the degree of immune activation. The HPLC method requires 100μL serum. Protein is removed by trichloroacetic acid. An external albumin-based calibrator is applied, and analysis is referred to an internal standard, 3-nitro-L-tyrosine. Kynurenine and nitrotyrosine are detected via UV absorbance at 360 nm wavelength, and tryptophan is detected via its natural fluorescence at 285nm extinction and 365 nm emission. Representative normal values of kynurenine and tryptophan were measured in the sera of 49 healthy blood donors.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Brown, R.R., Ozaki, Y., Datta, S.P., Borden, E.C., Sondel, P.M., and Malone, D.G., 1991, Implications of interferon-induced tryptophan catabolism in cancer, autoimmune diseases and AIDS, Adv. Exp. Med. Biol. 294:425–435.
Byrne, G.I., Lehmann, L.K., Kirschbaum, J.G., Borden, E.C., Lee, C.C.M., and Brown, R.R., 1986, Induction of tryptophan degradation in vitro and in vivo: A gamma interferon stimulated activity, J. Interferon Res. 6:389–396.
Denz, H., Orth, B., Weiss, G., Herrmann, R., Huber, P., Wachter, H., and Fuchs, D., 1993, Weight loss in patients with hematological neoplasias is associated with immune system stimulation, Clin. Investig. 71:37–41.
Fernstrom, J.D. and Wurtman, R.J., 1972, Brain serotonin content: physiological regulation by plasma neutral amino acids, Science 178: 414–416.
Freese, A., Swartz, K.J., During, M.J., and Martin, J.B., 1990, Kynurenine metabolites of tryptophan: Implications for neurologic diseases, Neurology 40:691–695.
Fuchs, D., Möller, A.A., Reibnegger, G., Stöckle, E., Werner, E.R., and Wachter, H., 1990, Decreased serum tryptophan in patients with HIV-1 infection correlates with increased serum neopterin and with neurologic/psychiatric symptoms, J. Acquir. Immune Def. Syndr. 3:873–876.
Fuchs, D., Möller, A.A., Reibnegger, G., Werner, E.R., Werner-Felmayer, G., Dierich, M.P., and Wachter, H., 1991, Increased endogenous interferon-gamma and neopterin correlate with increased degradation of tryptophan in human immunodeficiency virus type 1 infection, Immunol. Lett. 28:207–211.
Gasse, T., Murr, C., Meyersbach, P., Schmutzhard, E., Wachter, H., and Fuchs, D, 1994, Neopterin production and tryptophan degradation in acute neuroborreliosis versus late lyme encephalopathy, Eur. J. Clin. Chem. Clin. Biochem. 32:685–689.
Gasse, T., Widner, B., Baier-Bitterlich, G., Sperner-Unterweger, B., Leblhuber, F., Wachter, H., and Fuchs, D., 1998, Abnormal tryptophan metabolism, neurologic/psychiatric disturbances and its relationship to immune activation, in: Progress in HPLC-HPCE: Vol. 7. Neurochemical markers of degenerative diseases and drug addiction (G A. Qureshi, H. Parvez, P. Caudy, and S. Parvez, eds.), pp. 351–382, VSP Press, Zeist.
Haddad, I.Y., Pataki, G., Hu, P., Beckman, J.S., and Matalon, S., 1994, Quantitation of nitrotyrosine levels in lung sections of patients and animals with acute lung injury, J. Clin. Invest. 94:2407–2413.
Heyes, M.P., Saito, K., Crowley, J.S., Davis, L.E., Demitrack, M.A., Dez, M., Diling, L.A., Elia, J., Kruesi, M.J., and Lackner, A., 1992, Quinolinic acid and kynurenine pathway metabolism in inflammatory and non-inflammatory neurological disease, Brain 115:1249–1273.
Kaur, H. and Halliwell, B., 1994, Evidence for nitric oxide-mediated oxidative damage in chronic inflammation. Nitrotyrosine in serum and synovial fluid from rheumatoid patients, FEBS Lett. 350:9–12.
Launay, J.M., Copel, L., Callebert, J., Corvaia, N, Lepage, E., Bricaire, F., Saal, F., and Peries, J., 1988, Decreased whole blood 5-hydroxytryptamine (serotonin) in AIDS patients, J. Acquir. Immune Def. Syndr. 1:324–325.
Maes, M., Schärpe, S., Meltzer, H.Y., Okayli, G., Bosmans, E., D’Hondt, P., Vanden-Bossche, B.V., and Cosyns, P., 1994, Increased neopterin and interferon-gamma secretion and lower availability of L-tryptophan in major depression: further evidence for an immune response, Psychiatry Res. 54:143–160.
Milton, W.T. and Gensheng, F., 1991, Relationship between interferon-γ, indoleamine 2,3-dioxygenase, and tryptophan catabolism, FASEB J., 5:2516–2522.
Sayama, S., Yoshida, R., Oku, T., Imanishi, J., Kishida, T., and Hayaishi, O., 1981, Inhibition of interferonmediated induction of indoleamine 2,3-dioxygenase in mouse lung inhibitors of prostaglandin biosynthesis, Proc. Natl. Acad. Sci. USA 78:7327–7330.
Schmitz, J.L., Carlin, J.M., Borden, E.C., and Byrne, G.I., 1989, Beta-Interferon inhibites Toxoplasma gondii growth in human monocyte-derived macrophages, Infect. Immun. 57:3254–3256.
Werner, E.R., Bitterlich, G., Fuchs, D., Hausen, A., Reibnegger, G., Szabo, G., Dierich, M.P., and Wachter, H., 1987, Human macrophages degrade tryptophan upon induction by interferon-gamma, Life Sci. 41:273–280.
Widner, B., Werner, E.R., Schennach, H., Wachter, H., and Fuchs, D, 1997, Simultaneous measurement of serum tryptophan and kynurenine by HPLC, Clin. Chem. 43:2424–2426.
Yoshida, R., Imanishi, J., Oku, T., Kishida, T., and Hayaishi, O., 1981, Induction of pulmonary indoleamine 2,3-dioxygenase by interferon, Proc. Natl. Acad. Sci. USA 78:129–132.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Springer Science+Business Media New York
About this chapter
Cite this chapter
Widner, B., Werner, E.R., Schennach, H., Fuchs, D. (1999). An Hplc Method to Determine Tryptophan and Kynurenine in Serum Simultaneously. In: Huether, G., Kochen, W., Simat, T.J., Steinhart, H. (eds) Tryptophan, Serotonin, and Melatonin. Advances in Experimental Medicine and Biology, vol 467. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-4709-9_105
Download citation
DOI: https://doi.org/10.1007/978-1-4615-4709-9_105
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-7133-5
Online ISBN: 978-1-4615-4709-9
eBook Packages: Springer Book Archive