Clinical Chemistry 46: 112-117, 2000;
(Clinical Chemistry. 2000;46:112-117.)
© 2000 American Association for Clinical Chemistry, Inc.
Enzyme Immunoassay for Autoantibodies to Human Liver-Type Arginase and Its Clinical Application
Masahiro Kimura1,
Ke-Ita Tatsumi1,
Hisato Tada1,
Masaki Ikemoto2,
Yoshihiro Fukuda2,
Akira Kaneko3,
Michio Kato3,
Yoh Hidaka1 and
Nobuyuki Amino1,a
1
Department of Laboratory Medicine, Osaka University Medical School, Osaka 565-0871, Japan.
2
College of Medical Technology, Kyoto University, Kyoto
606-8507, Japan.
3
Department of Gastroenterology, Osaka National Hospital,
Osaka 540-0006, Japan.
a Address correspondence to this author at: Department of Laboratory Medicine, Osaka University Medical School D2, Osaka 565-0871, Japan. Fax 81-6-6879-3239; e-mail namino{at}labo.med.osaka-u.ac.jp
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Abstract
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Background: Arginase is an enzyme of the urea cycle, and one of
the two isoenzymes is the liver-type enzyme. We examined serum
autoantibodies to this liver-type enzyme in patients with hepatitis.
Methods: Antibodies to recombinant human liver-type arginase were
measured by ELISA in 95 patients and 55 healthy controls.
Results: The mean absorbance values in the ELISA assays of
patients with definite autoimmune hepatitis (n = 11;
P <0.0001), probable autoimmune hepatitis (n = 31;
P <0.0001), and hepatitis C (HCV; n = 20;
P <0.01) were significantly different from those of
healthy controls, but the values in patients with hepatitis B (HBV;
n = 23) and other autoimmune diseases (n = 10) were not
significantly different from those of healthy controls. When the cutoff
was fixed at the upper 95th percentile of the absorbance value in
healthy controls, positive reactions were found in 18.2%, 32.3%,
20.0%, 13.0%, and 10.0% of patients with definite autoimmune
hepatitis, probable autoimmune hepatitis, HCV hepatitis, HBV hepatitis,
and other autoimmune diseases, respectively. All of these positive
reactions were abolished by inhibition of serum with recombinant
antigen. The specificity and sensitivity of this ELISA were 96% and
29%, respectively. The intraassay and interassay coefficients of
variation were 2.3–7.5% and 9.8–11%, respectively. There
was no relationship between these antibodies and anti-nuclear,
anti-smooth muscle, or anti-cytochrome P450IID6 antibodies.
Conclusions: The ELISA for anti-liver-type arginase autoantibody
improved the detectability of autoimmune hepatitis when compared with
established assays for liver-specific autoantibodies.
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Introduction
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Autoimmune hepatitis
(AIH)1
has been recognized for >30 years, but its diagnostic
criteria have only recently been codified (1). To support
the diagnosis of AIH, several laboratory tests for autoantibodies, such
as anti-nuclear antibodies, anti-smooth muscle antibodies, liver-kidney
microsome antibodies, anti-neutrophil cytoplasmic antibodies, and
antibodies to soluble liver antigen, have been used (2)(3)(4).
AIH has been subdivided into three types, based on the different
patterns of these antibodies (1)(5). However,
most of these antibodies are not organ specific (2) and are
unsatisfactory for routine use. Therefore, we searched for a
liver-specific antigen and tried to establish an ELISA for
liver-specific autoantibodies.
Arginase (EC 3.5.3.1; L-arginine amidinohydrolase)
catalyzes hydrolysis of arginine to urea and ornithine in the liver of
ureotelic animals. Two forms of arginase have been defined: liver-type
arginase (arginase 1) and extrahepatic type arginase (arginase 2)
(6)(7). Liver-type arginase differs from
extrahepatic type arginase in immunological properties and enzyme
kinetics, and it is quite abundant in the liver cytosol. cDNAs encoding
human liver-type arginase (8)(9) and
extrahepatic arginase (10) have been cloned from human liver
and kidney, respectively. Liver-type arginase mRNA is expressed
strongly in the liver, whereas extrahepatic arginase mRNA is
expressed strongly in the adult kidney and is present in lesser amounts
in other extrahepatic tissues (11)(12).
In 1985, Mafune et al. (13) reported that rabbit antisera
against rat liver arginase showed direct cytotoxic effects and
antibody-dependent cell-mediated cytotoxicity against primary cultured
hepatocytes of rat. They suggested that autoantibodies to liver-type
arginase may have an important role in the pathogenesis of AIH. In this
study, we developed an ELISA that uses recombinant human liver-type
arginase as an antigen for the detection of anti-arginase
autoantibodies in patients with AIH.
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Materials and Methods
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patients
Serum samples were obtained from 11 patients with definite AIH, 31
patients with probable AIH, 20 patients with hepatitis C (HCV), 23
patients with hepatitis B (HBV), 10 patients with other autoimmune
diseases, and 55 healthy controls. We obtained informed consent from
all patients. The mean ages and sex distribution are summarized in
Table 1
. The mean ages of the various patient groups were not
significantly different from those of the controls. Diagnosis of AIH
was made by the scoring system of the International Autoimmune
Hepatitis Group (1). Definite AIH patients had scores 
16,
and probable AIH patients had scores of 10–15 before treatment. Two
patients with definite AIH and two other patients with probable AIH had
been receiving steroid therapy at the time of serum sampling, but the
rest of the patients had received no immunosuppressive therapy before
venesection. Two of the 31 patients with probable AIH had HCV
infections. HBV infection was diagnosed by positive reaction in an
enzyme immunoassay (Abbott AxSYM HBsAg; Abbott Diagnostics) for
hepatitis B surface antigen. HCV hepatitis was diagnosed by
positive reaction for HCV antibody, measured by enzyme immunoassay
(Ortho HCV Ab ELISA Test III; Ortho Clinical Diagnostics). One of 20
patients with HCV hepatitis also had a HBV infection.
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Table 1. Age, sex, and anti-arginase antibodies in patients with
hepatitis and autoimmune diseases, and healthy
controls.
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preparation of recombinant human liver-type arginase
Escherichia coli K-12 strain KY1436 was transformed by
pTAA12 expression plasmid vector containing human liver-type arginase
cDNA. Expression of the recombinant human arginase was induced with
isopropyl ß-D-thiogalactopyranoside. The cells
were harvested by centrifugation and were resuspended in an adequate
volume of 10 mmol/L Tris-HCl buffer, pH 7.4. The suspended cells were
sonicated on ice in short bursts and centrifuged. To obtain purified
recombinant human arginase from the supernatant, we used CM-Sephadex
C-50 (Amersham Pharmacia Biotech), DEAE-cellulose (DE52;
Whatman), and Sephadex G-150 columns according to the previously
reported method (14).
elisa procedure
Microtiter plates (Maxisorp; Nunc) were coated overnight at
4 °C with recombinant human liver-type arginase dissolved in
coating buffer (0.1 mol/L
NaH2PO4-NaOH, 1 g/L
NaN3, pH 7.5) at 20 mg/L (100 µL per
well). The plates were subsequently washed twice with coating buffer
and incubated with 200 µL of blocking solution (coating buffer
containing 30 g/L bovine serum albumin) for 2 h at
37 °C. After the plates were washed twice with washing solution A
(20 mmol/L Tris-HCl, 500 mmol/L NaCl, 0.5 mL/L Tween 20, pH
7.5) and once with washing solution B (20 mmol/L Tris-HCl, 500 mmol/L
NaCl, pH 7.5), they were dried, sealed, and stored at 4 °C.
Each well of a coated microtiter plate was filled with 100 µL of
serum sample diluted 1:100 in reaction solution (washing solution A
containing 1 g/L bovine serum albumin) and incubated at 25 °C for
1 h. The plate was washed five times with washing solution A, and
each well was incubated with 100 µL of protein A coupled with
horseradish peroxidase (diluted 1:10 000; Amersham Pharmacia
Biotech). After another 1-h incubation at 25 °C, the plate was
washed three times with washing solution A and twice with washing
solution B. The horseradish peroxidase activity retained in the wells
was assayed by addition of 100 µL of 3,3',5,5'-tetramethylbenzidine
substrate solution (Kirkegaard & Perry Laboratories). After 30 min at
25 °C, the reaction was stopped by the addition of 100 µL of stop
solution containing 10 mL/L HCl (Kirkegaard & Perry) to each well, and
the absorbance was measured within 30 min at 450 nm by a microtiter
plate reader (MPR-A4; TOSOH). Nonspecific background absorbance
obtained with pooled serum from healthy subjects was subtracted
from each value.
inhibition study
Inhibition studies were performed according to the method of Ma et
al. (15) with slight modifications. In brief, strips of
polyvinylidene difluoride membrane (1.0 x 1.0 cm,
Hybond-P; Amersham Pharmacia Biotech) were impregnated with 50 µg of
recombinant human arginase. Residual nonspecific reactivity of the
strips was then blocked by incubation in blocking solution for 1 h
at room temperature. The strips were incubated in patient sera diluted
1:100 in reaction solution for 2 h at room temperature; the
procedure was performed in triplicate. As a control, nonimpregnated
strips were incubated with diluted serum. Sera preincubated with
impregnated or nonimpregnated strips were tested for reactivity against
recombinant human arginase by ELISA.
detection of anti-cytochrome p450iid6 antibody
Anti-cytochrome P450IID6 (CYP2D6) antibodies were detected by a
radioligand-binding assay, according to the method reported previously
(16)(17). In brief,
[35S]methionine-labeled recombinant human
CYP2D6 was produced by in vitro transcription and translation using a
TNT-coupled reticulocyte lysate system (Promega). Patient sera
(diluted 1:50) and labeled human CYP2D6 were incubated overnight at
4 °C. The [35S]methionine-labeled
recombinant human CYP2D6-antibody complexes were precipitated with
protein A-Sepharose 4FF (Amersham Pharmacia Biotech) for 45 min at room
temperature. The quantity of precipitated, labeled CYP2D6 was
determined by measuring the radioactivity with a 1450 Micro Beta TRILUX
apparatus (Amersham Pharmacia Biotech). The results were expressed as a
CYP2D6 index calculated as below:
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other assays
Anti-nuclear and anti-smooth muscle antibodies were measured by
indirect immunofluorescence assays using commercial kits: the
Fluorohepana Test (Medical and Biological Laboratories) for
anti-nuclear antibodies and the Fluoro AID-1 Test (Medical and
Biological Laboratories) for anti-smooth muscle antibodies.
statistical analysis
The Mann–Whitney U-test was used to compare absorbance
readings of the patient groups and the control group. P
<0.05 was considered significant. Pearson correlation coefficient
analysis was used to examine the correlation between the titers of
anti-arginase antibodies and other indicators.
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Results
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dilution curve
Sera obtained from three patients with AIH with positive
anti-arginase antibodies were diluted in reaction solution. Eight
dilutions were obtained by serial 1:1 dilution of a serum from 1:50.
Values are given as means of duplicate determinations. The dilution
curve was linear in the dilution range between 50- and 800-fold (Fig. 1
). Therefore, we decided to use a serum dilution of 1:100 in
subsequent experiments.
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Figure 1. Serum dilution curve in AIH patients with positive
anti-liver-type arginase antibodies (•) and in healthy controls
( ).
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precision
In the ELISA for anti-liver-specific arginase antibodies, the
within-run coefficient of variation (CV) was 2.3% (n = 7) for
sera with high antibody concentrations (mean absorbance, 1.82) and
7.5% (n = 7) for sera with medium antibody concentrations (mean
absorbance, 1.11). The between-run CV was 11% (n = 7) for
sera with high antibody concentrations and 9.8% (n = 7) for sera
with medium antibody concentrations.
inhibition study
To exclude the possibility of nonspecific reactions in the ELISA
method, an inhibition study was performed. The absorbance values at 450
nm of the serum samples containing positive antibodies were markedly
decreased (>45%) after inhibition with recombinant human liver-type
arginase (Fig. 2
). In contrast, absorbance values were unchanged after
inhibition with control membranes that were not impregnated with
arginase. Positive antibody reactions in the non-AIH patients were also
completely abolished by inhibition with liver-type arginase
antigen.
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Figure 2. Serum inhibition by recombinant human liver-type arginase
in the anti-arginase antibody ELISA.
Without, no recombinant human liver-type arginase;
With, recombinant human liver-type arginase added.
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anti-liver-type arginase antibodies in various diseases
Using this ELISA, we examined serum samples obtained from patients
and compared the results with those of healthy controls. As shown in
Table 1
, the mean absorbance at 450 nm was significantly higher in
patients with AIH, either definite or probable, than in healthy
controls; the absorbance at 450 nm was also higher in HCV hepatitis
patients, but not patients with HBV hepatitis or other autoimmune
diseases, than in healthy controls.
Individual absorbance values are shown in Fig. 3
. The upper 95th percentile limit of the absorbance value in
healthy controls was calculated by a parametric method after
square-root-power transformation of the absorbance value. Values
greater than this limit (0.640 at 450 nm) were considered
positive. As described above, these positive values became negative
after inhibition with liver-type arginase in all samples. Accordingly,
18.2%, 32.3%, 20.0%, 13.0%, and 10.0% of patients were positive
for definite AIH, probable AIH, HCV hepatitis, HBV hepatitis, and other
autoimmune diseases, respectively. Three of 55 healthy controls and 12
of 42 total patients with AIH had positive anti-arginase antibodies.
Thus, the specificity and sensitivity of this ELISA were 96% and 29%,
respectively.
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Figure 3. Individual absorbance values in the anti-human liver-type
arginase antibody ELISA of samples from patients with AIH, HCV
hepatitis, HBV hepatitis, and other autoimmune diseases.
The dashed line shows the upper 95th percentile limit of
the absorbance values (0.640 at 450 nm) in healthy controls
(Normal).
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relationship between anti-arginase antibodies and other
indicators
Anti-arginase antibody titers were compared with serum IgG
concentrations (Fig. 4
A), anti-nuclear antibody titers (Fig. 4B
), and anti-CYP2D6
antibody activity (Fig. 4C
) in patients with AIH. None of these
indicators was related to the anti-arginase antibody titer. Definitive
positive antibody reaction against CYP2D6 was observed in only 2 of 42
(4.8%) patients with AIH. The anti-arginase antibody activity was not
related to the presence of anti-smooth muscle antibodies or increased
serum concentrations of aspartate aminotransferase or alanine
aminotransferase. No relationship was observed between positive
anti-arginase antibodies and a family history of AIH or other
autoimmune diseases. There were no special clinical characteristics in
the patients positive for anti-arginase antibodies.
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Figure 4. Relationship between the anti-human liver-type arginase
antibody activity and the serum concentration of IgG
(A), the anti-nuclear antibody titer (B),
or the anti-CYP2D6 antibody activity (C) in patients
with AIH.
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Discussion
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Excellent laboratory tests for autoantibodies, such as
anti-thyroid microsomal antibody, anti-thyroid-stimulating hormone
receptor antibody, and anti-glutamic acid decarboxylase antibody,
enable us to screen for subclinical autoimmune diseases and to predict
the clinical onset of disease (18)(19). Thus, we
tried to establish a new autoantibody test for specific detection of
hepatic autoimmune abnormalities.
Among several autoantigens in the liver, human liver-type arginase
(arginase 1) is specifically expressed in liver cells. Our search of
the medical literature revealed no report on autoantibodies to human
liver-type arginase. Therefore, we tried to develop an ELISA for
anti-human liver-type arginase antibodies, and we succeeded in
establishing a methodologically satisfactory ELISA system.
Unexpectedly, however, the prevalence of positive reactions was not
high enough to detect definite AIH. However, one-third of patients with
probable AIH showed positive reactions. This result was not induced by
immunosuppressive therapy because only 4 of 42 patients with AIH (2
definitive, 2 probable) were receiving immunosuppressive therapy at the
time of serum sampling. In some autoimmune diseases, autoantibodies
gradually disappear according to the progress of disease
(20). Our data in this study suggest that anti-arginase
antibodies might be detectable in mild forms of AIH, although the
difference in antibody occurrence was not statistically significant
between the two groups.
Many studies have reported on non-organ-specific autoantibodies;
however, little is known about the frequency of autoantibodies to
liver-specific antigen(s) within patients with AIH. Only 4.8% of
patients with AIH in this study had definitive positive antibodies to
CYP2D6. Anti-CYP2D6 antibodies usually are found in patients with type
2 AIH, who constitute 5% of all AIH patients (21). In
previous studies, the prevalence of CYP2D6 antibodies in patients with
AIH was reported as <5% (22)(23).
Antibodies to soluble liver antigen were found in only 11% of patients
with AIH (22). Autoantibodies to asialoglycoprotein
receptor, which is expressed on the surface of hepatocytes, were found
in 50–80% of patients with AIH (24)(25).
However, positive antibodies were also found in 6.5–73.0% of patients
with active HBV hepatitis (24)(25). Thus, the
specificity of anti-asialoglycoprotein-receptor antibodies toward AIH
apparently is low (25). Among all AIH patients, therefore,
it is clear that this ELISA for anti-arginase antibodies is far better
than the established assay for anti-CYP2D6 antibody or soluble
liver antigen from the point of view of screening for autoimmune
hepatic abnormality.
It has been speculated that AIH may be triggered by infection with
hepatotropic viruses, and the relationship between HCV infection and
AIH is of interest (26). Several patients with HCV or HBV
infection had positive anti-arginase antibodies in this study. This may
not have been attributable to nonspecific reactions in the ELISA
because the positive reactions became negative after specific
inhibition with human liver-type arginase antigen. According to the
scoring system of the International Autoimmune Hepatitis Group, the
presence of viral infection subtracts two or three points from the
total score; thus, patients who had both AIH and viral infections might
easily be dropped from the group with AIH. Actually, in this study, no
patients with definite AIH had hepatitis B surface antigen or HCV
antibody. Further study is needed to clarify the clinical significance
of positive antibodies in patients with viral hepatitis.
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Acknowledgments
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This work was supported by grants-in-aid for Scientific Research
from the Ministry of Education, Science and Culture of Japan
(Grants-in-Aid 09307055 and 11357021 to N.A. and Grant-in-Aid 09772066
to H.T.). We greatly appreciate the valuable comments of Drs. Manabu
Masuzawa and Kiyoshi Ichihara. We also thank Akiko Izumi for
skillful assistance.
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Footnotes
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1 Nonstandard abbreviations: AIH, autoimmune hepatitis; HCV, hepatitis C virus; HBV, hepatitis B virus; and CYP2D6, cytochrome P450IID6. 
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References
|
|---|
-
Johnson PJ, McFarlane IG. Meeting report: International Autoimmune Hepatitis Group. Hepatology 1993;18:998-1005.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Krawitt EL. Autoimmune hepatitis. N Engl J Med 1996;334:897-903.
[Free Full Text]
-
Glass RD, Knox WE. Arginase isozymes of rat mammary gland, liver, and other tissues. J Biol Chem 1973;248:5785-5789.
[Abstract/Free Full Text]
-
Czaja AJ. The variant forms of autoimmune hepatitis. Ann Intern Med 1996;125:588-598.
[Abstract/Free Full Text]
-
van den Berg AP. Autoimmune hepatitis: pathogenesis, diagnosis and treatment. Scand J Gastroenterol 1998;33(Suppl 225):66-69.
-
Kaysen GA, Strecker HJ. Purification and properties of arginase of rat kidney. Biochem J 1973;133:779-788.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Spector EB, Rice SC, Cederbaum SD. Immunologic studies of arginase in tissues of normal human adult and arginase-deficient patients. Pediatr Res 1983;17:941-944.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Dizikes GJ, Grody WW, Kern RM, Cederbaum SD. Isolation of human liver arginase cDNA and demonstration of nonhomology between the two human arginase genes. Biochem Biophys Res Commun 1986;141:53-59.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Haraguchi Y, Takiguchi M, Amaya Y, Kawamoto S, Matsuda I, Mori M. Molecular cloning and nucleotide sequence of cDNA for human liver arginase. Proc Natl Acad Sci U S A 1987;84:412-415.
[Abstract/Free Full Text]
-
Gotoh T, Sonoki T, Nagasaki A, Terada K, Takiguchi M, Mori M. Molecular cloning of cDNA for nonhepatic mitochondrial arginase (arginase II) and comparison of its induction with nitric oxide synthase in a murine macrophage-like cell line. FEBS Lett 1996;395:119-122.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Gotoh T, Araki M, Mori M. Chromosomal localization of the human arginase II gene and tissue distribution of its mRNA. Biochem Biophys Res Commun 1997;233:487-491.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Morris SM, Jr, Bhamidipati D, Kepka-Lenhart D. Human type II arginase: sequence analysis and tissue-specific expression. Gene 1997;193:157-161.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Mafune N, Ideta N, Watabe H, Nagura H, Kobayashi K. Occurrence of cytotoxic autoantibody in rabbits by immunization with heterologous liver arginase: a possible implication in the mechanism of the autoimmune liver diseases. Clin Exp Immunol 1985;59:123-131.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Ikemoto M, Tabata M, Miyake T, Kono T, Mori M, Totani M, et al. Expression of human liver arginase in Escherichia coli. Purification and properties of the product. Biochem J 1990;270:697-703.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Ma Y, Gaken J, McFarlane BM, Foss Y, Farzaneh F, McFarlane IG, et al. Alcohol dehydrogenase: a target of humoral autoimmune response in liver disease. Gastroenterology 1997;112:483-492.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Ma Y, Gregorio G, Gaken J, Muratori L, Bianchi FB, Mieli-Vergani G, et al. Establishment of a novel radioligand assay using eukaryotically expressed cytochrome P4502D6 for the measurement of liver kidney microsomal type antibody in patients with autoimmune hepatitis and hepatitis C virus infection. J Hepatol 1997;26:1396-1402.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Yamamoto AM, Johanet C, Duclos-Vallee JC, Bustarret FA, Alvarez F, Homberg JC, et al. A new approach to cytochrome CYP2D6 antibody detection in autoimmune hepatitis type-2 (AIH-2) and chronic hepatitis C virus (HCV) infection: a sensitive and quantitative radioligand assay. Clin Exp Immunol 1997;108:396-400.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Hidaka Y, Tamaki H, Iwatani Y, Tada H, Mitsuda N, Amino N. Prediction of post-partum Graves’ thyrotoxicosis by measurement of thyroid stimulating antibody in early pregnancy. Clin Endocrinol 1994;41:15-20.
[Medline]
[Order article via Infotrieve]
-
Tuomilehto J, Zimmet P, Mackay I, Koskela P, Vidgren G, Toivanen L, et al. Antibodies to glutamic acid decarboxylase as predictors of insulin-dependent diabetes mellitus before clinical onset of disease. Lancet 1994;343:1383-1385.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Ohta M, Obayashi H, Takahashi K, Kitagawa Y, Nakano K, Matsuo S, et al. Radioimmunoprecipitation assay for glutamic acid decarboxylase antibodies evaluated clinically with sera from patients with insulin-dependent diabetes mellitus. Clin Chem 1996;42:1975-1978.
[Abstract/Free Full Text]
-
Meyer zum Buschenfelde KH, Dienes HP. Autoimmune hepatitis. Virchows Arch 1996;429:1-12.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Czaja AJ, Carpenter HA, Manns MP. Antibodies to soluble liver antigen, P450IID6, and mitochondrial complexes in chronic hepatitis. Gastroenterology 1993;105:1522-1528.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Nishioka M, Morshed SA, Kono K, Himoto T, Parveen S, Arima K, et al. Frequency and significance of antibodies to P450IID6 protein in Japanese patients with chronic hepatitis C. J Hepatol 1997;26:992-1000.
[Web of Science][Medline]
[Order article via Infotrieve]
-
McFarlance BM, McSorley CG, Vergani D, McFarlane IG, Williams R. Serum autoantibodies reacting with the hepatic asialoglycoprotein receptor protein (hepatic lectin) in acute and chronic liver disorders. J Hepatol 1986;3:196-205.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Treichel U, Poralla T, Hess G, Manns M, Meyer zum Buschenfelde KH. Autoantibodies to human asialoglycoprotein receptor in autoimmune-type chronic hepatitis. Hepatology 1990;11:606-612.
[Web of Science][Medline]
[Order article via Infotrieve]
-
Mackie FD, Peakman M, Yun M, Sallie R, Smith H, Davis ET, et al. Primary and secondary liver/kidney microsomal autoantibody response following infection with hepatitis C virus. Gastroenterology 1994;106:1672-1675.
[Web of Science][Medline]
[Order article via Infotrieve]
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Top
Abstract
Introduction
