title: Determination of the serum metallothionein (MT)1/2 concentration in patients with Wilson's disease and Menkes disease.
abstract: We have developed an easy and specific enzyme-linked immunoassay (ELISA) for the simultaneous determination of serum metallothinein-1 (MT-1) and 2 (MT-2) in both humans and experimental animals. A competitive ELISA was established using a specific polyclonal antibody against rat MT-2. The antibody used for this ELISA had exhibited the same cross-reactivity with MT in humans and experimental animals. The NH2 terminal peptide of MT containing acetylated methionine was shown to be the epitope of this antibody. The reactivity of this ELISA system with the liver, kidney and brain in MT1/2 knock-out mice was significantly low, but was normal in an MT-3 knock-out mouse. The lowest detection limit of this ELISA was 0.6ng/ml and the spiked MT-1was fully recovered from the plasma. We investigated the normal range of MT1/2 (25-75%tile) in 200 healthy human serum and found it to be 27-48ng/ml, and this was compared with the serum levels in various liver diseases. The serum MT1/2 levels in chronic hepatitis C (HCV) patients were significantly lower than healthy controls and also other liver diseases. In the chronic hepatitis cases, the MT1/I2 levels increased gradually, followed by the progression of the disease to liver cirrhosis and hepatocellular carcinoma. In particular, we found significantly elevated MT1/2 plasma levels in Wilson's disease patients, levels which were very similar to those in the Long-Evans Cinnamon (LEC) rat (model animal of Wilson's disease). Furthermore, a significantly elevated MT1/2 level was found in patients with Menkes disease, an inborn error of copper metabolism such as Wilson's disease.
authors.affiliation: Graduate School of Health Sciences, Gunma University, Maebashi, Gunma, Japan.
authors.forename: Kyoumi
authors.initials: K
authors.lastname: Nakazato
authors.affiliation: Graduate School of Health Sciences, Gunma University, Maebashi, Gunma, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Tomioka
authors.affiliation: Graduate School of Health Sciences, Gunma University, Maebashi, Gunma, Japan. Electronic address: nakajimak05@ybb.ne.jp.
authors.forename: Katsuyuki
authors.initials: K
authors.lastname: Nakajima
authors.affiliation: Frontier Institute, Ishikari, Hokkaido, Japan.
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Saito
authors.affiliation: Frontier Institute, Ishikari, Hokkaido, Japan.
authors.forename: Mihoko
authors.initials: M
authors.lastname: Kato
authors.affiliation: Frontier Institute, Ishikari, Hokkaido, Japan.
authors.forename: Tsukasa
authors.initials: T
authors.lastname: Kodaira
authors.affiliation: Diabetes and Metabolic Disease Research Center, Hidaka Hospital, Takasaki, Japan.
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Yatsuzuka
authors.affiliation: Diabetes and Metabolic Disease Research Center, Hidaka Hospital, Takasaki, Japan.
authors.forename: Younosuke
authors.initials: Y
authors.lastname: Shimomura
authors.affiliation: Diabetes and Metabolic Disease Research Center, Hidaka Hospital, Takasaki, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Hiroki
authors.affiliation: Department of Pediatrics, Teikyo University School of Medicine, Tokyo, Japan.
authors.forename: Kahoko
authors.initials: K
authors.lastname: Motoyama
authors.affiliation: Department of Pediatrics, Teikyo University School of Medicine, Tokyo, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Kodama
authors.affiliation: Graduate School of Health Sciences, Gunma University, Maebashi, Gunma, Japan.
authors.forename: Takeaki
authors.initials: T
authors.lastname: Nagamine
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006527:Hepatolenticular Degeneration / Q000097:blood*; D006801:Humans; D008297:Male; D007706:Menkes Kinky Hair Syndrome / Q000097:blood*; D008668:Metallothionein / Q000097:blood*; D000090023:Metallothionein 3; D018345:Mice, Knockout; D008875:Middle Aged; D055815:Young Adult
chemical_list: D000090023:Metallothionein 3; C000715430:Mt3 protein, mouse; D008668:Metallothionein
keywords: Liver disease; Long–Evans Cinnamon (LEC) rat; Menkes disease; Metallothionein 1/2; Wilson's disease