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Transforming growth factor-β1 (TGF-β1) in chronic hepatitis C
336A 917
AASLD
DEFECTIVE ANTMRAL CYTOKINES lN CHRONIC HEPATITIS C VIRUS-INFECTED LIVERS. AS Gaweco’. V Rust&. T Hassaneh?, L El-Asbmaw?. T Whiteside’. WJ Hofinann’, DH Van Tbiel’. HF Otto’. Inst. of Pathology’, Univ. of Heidelberg, Germany, INOVA Inst2, Oklahoma Transplant Inst.‘, & Dept. of Pathology4, Univ. of Pittsburgh, USA
9 18
The pathogenesis of chronic HCV infection remains elusive. Host viral immunity involves effector mechanisms critically regulated by elaborated cytokines, some of which exhibit potent antiviral properties. A systemic defect in the production of several antiviral cytokines have been described, however, the preferential compartmentalization of HCV-specific immune effector cells necessitates a tissue-specific description of cytokine dysregulation at the site of liver injury. Methods: Semi-quantitative RT-PCR was performed in liver biopsies of 24 chronic HCV-infected patients. The mRNA expression of IL-lb, IL-6, IL-8, TNF-a and IL-2R a-chain were analyzed and compared to 17 normal liver tissue samples. In siti hybridization using “S-labeled cDNA probes for IL-6 and TNFa was performed separately .on liver biopsies of 4 chronic HCV-infected and 3 control patients with normal histology and liver function. All chronic HCV patients were seropositive for anti-HCV and HCVRNA PCR and had persistently elevated ALT levels >6 months and chronic hepatitis histology. All control samples were HCV-RNA PCR negative.
chronic HCV-infected livers associated with a downregulated IL-2Ro. (p=O.O037) expression using RT-PCR, in contrast to its constitutive expression in 17 normal liver samples. In situ hybridization analysis of additional liver biopsies demonstrated conspicuous TNFa expression in 2/3 of control patients, whereas only few TNF-a mRNA expressing cells were detected in 2/4 of chronic HCV-infected patients. Conclusion: The defective gene activation of the monocyte/macropbage-associated antiviral cytoldnes demonstrates another immunological defect complementing known impaired 1FN-a in chronic HCV-infected livers, probably as a result of direct of mononuclear
cell function
and cytokine
inhibition
CHANGES
IN SERUM SOLUBLE INTERLEUKIN 2 RECEPTOR LEVELS WITH CHRONIC HEPATITIS C BY INTERFERON-a. T. Takeda. S. Nishiwchi. S. Nakaiima. S. Shiomi. T. Kuroki. K, Third Department of Internal Medicine, Osaka City Kobavashi. University Medical School, Osaka, Japan.
IN PATIENTS
known
hepatitis.
Serum soluble interleukin to increase in patients
We
examined
the
2 receptor
(sIL2R)
levels
with acute and chronic relationship between sIL2R
are
viral and
eff’icaqy of interferon (IFN) treatment for patients with chronic hepatitis C (CH-C). We examined the time course changes in sIL2R in sera of patients during and 12 months after treatment. Human lymphoblastoid IFN was injected (im) at a dose of 6 Mega U/day every day for 2 weeks, and three times per week for 10 to 22 weeks. Serum sIL2R levels were measured using an enzyme immunoassay. The ratio of serum sIL2R at each time point Lo that before treatment was used to examine changes in sIL2R levels.
Fifty-nine
patients
with
CH-C
were
studied.
They
included
29
patients whose serum continued to be positive for HCVRNA after treatment (NR group). And 30 patients in whom HCVRNA was eliminated from serum and continued 10 be negative (CR group). In the patients, slL2R levels were correlated with ALT levels (r=O.465, p
1995
QUANTITATIVE ANALYSIS OF INTERFERON o / p RECEPTOR mRNA IN THE LIVER OF PATIENTS WITH CHRONIC HEPATITIS C H.Yatsuhashi,MC.Parauet.K.Yama&i,F.Iio.T.Aritomi.M.Yamasaki, O.Inoue.M.Kopa.M.Yano. Institute for Clinical Research, Nagasaki Chuo National Hospital, Nagasaki ,Japan.
association
by the
and response
hepatitis C v$us.
919
October
Background and methods:IFN a /,l3 receptor identified by Novick (1994) binds and responds effectively to IFN /3 and to several IFN (Y subtypes. But there has been no report on IFN (Y/P receptor associated with liver disease.We used RT-PCR method for the relative quantification of gene expression using a simultaneously amplified sequence of /3-a& mRNA as a internal control for the target sequence of IFN a /,L? receptor mRNA in the liver of 29 patients with chronic hepatitis C and in 6 controls with fatty liver.The PCR product of the internal control was reduced by delaying the addition of the primers for p-a&n. The cycle differences between the logarithmic phase of the curves for target sequence and for the p actin(&ycle) measured with a densitometry indicated the level of IFN (I /,8 receptor mRNA. Results:IFN a / p receptor mRNA in all patients and conlrols were detected. In chronic hepatitis C the mean concentration of IFN a / ,8 receptor mRNA was 2.0 fold higher than in the controls. There was no correlation between mRNA levels and ALT levels or histological findings in chronic hepatitis C. There was a significant reverse correlation between mRNA levels and serum HCV-RNA levels measured by b-DNA probe assay.(r=0.5589 P
Results: mRNA transcripts for IL-Q3 @=0.00079), IL-6 (p=O.O0017), IL-8 (p=O.O017), and TNFa @=0.00014) were detected in low numbers in 24
impairment
HEPATOLOGY
ABSTRACTS
920
with
biological
activity
of IFN
for IFN treatment in chronic
to reduce
viral replication
hepatitis C .
TRANSFORMING GROWTH FACTOR431 (TGFM) IN CHRONIC HEPATITIS C Nelson DR. Marousis CG. Oian RP. Xu Y. Gonzalez-Per&a RP. Davis CL, Lau JYN. Section of Hepatobiiary Diseases, University of Florida, Gainesville, FL. Baclmrouod: TGFal has been implicated in mediating hepatic tibrogenesis. TGP t31 is also known to have. negative regulatory effects on the immune system. Serum Th2 cytokines are elevated in chronic HCV infection. Hvuothesis: TGF-Bl production is increased ia chronic HCV infection. Sweitic Alms: (1) To determine the serum levels of TGFDl (total and biologically active) in chronic HCV infection and their clinical implications. (2) To study the relationship of TGFal with Th2 cytokiues (IL4, IL-10).&&&z Serum samples were prospectively collected on 88 chronic HCV patients (M:F 49:39, age 23-68) and 22 healthy controls (M:F U:9 age 19-63). Total (using acid activation) and biologically active TGFa 1, IL4, and IL10 were measured by ELISA. HCV RNA levels were quantitated by bDNA (Chiron) and genotypes were determined by RFLP based oa SUTR. Histological diagnosis was available in.87 patients (54 sections were available for complete analysis for disease activity). Liver sections from 80 other HCV patients were evaluated for hepatic expression of TGFal using immuaohistochemistry. m: HCV patients had a higher level of TGFal, both total (817f 464 rig/ml) and the biologically active form (52Or 370 pg/ml, 0=88), compared to controls (n=U, total TGFal: 183i 105 w/ml. n
AASLD
DEFECTIVE ANTMRAL CYTOKINES lN CHRONIC HEPATITIS C VIRUS-INFECTED LIVERS. AS Gaweco’. V Rust&. T Hassaneh?, L El-Asbmaw?. T Whiteside’. WJ Hofinann’, DH Van Tbiel’. HF Otto’. Inst. of Pathology’, Univ. of Heidelberg, Germany, INOVA Inst2, Oklahoma Transplant Inst.‘, & Dept. of Pathology4, Univ. of Pittsburgh, USA
9 18
The pathogenesis of chronic HCV infection remains elusive. Host viral immunity involves effector mechanisms critically regulated by elaborated cytokines, some of which exhibit potent antiviral properties. A systemic defect in the production of several antiviral cytokines have been described, however, the preferential compartmentalization of HCV-specific immune effector cells necessitates a tissue-specific description of cytokine dysregulation at the site of liver injury. Methods: Semi-quantitative RT-PCR was performed in liver biopsies of 24 chronic HCV-infected patients. The mRNA expression of IL-lb, IL-6, IL-8, TNF-a and IL-2R a-chain were analyzed and compared to 17 normal liver tissue samples. In siti hybridization using “S-labeled cDNA probes for IL-6 and TNFa was performed separately .on liver biopsies of 4 chronic HCV-infected and 3 control patients with normal histology and liver function. All chronic HCV patients were seropositive for anti-HCV and HCVRNA PCR and had persistently elevated ALT levels >6 months and chronic hepatitis histology. All control samples were HCV-RNA PCR negative.
chronic HCV-infected livers associated with a downregulated IL-2Ro. (p=O.O037) expression using RT-PCR, in contrast to its constitutive expression in 17 normal liver samples. In situ hybridization analysis of additional liver biopsies demonstrated conspicuous TNFa expression in 2/3 of control patients, whereas only few TNF-a mRNA expressing cells were detected in 2/4 of chronic HCV-infected patients. Conclusion: The defective gene activation of the monocyte/macropbage-associated antiviral cytoldnes demonstrates another immunological defect complementing known impaired 1FN-a in chronic HCV-infected livers, probably as a result of direct of mononuclear
cell function
and cytokine
inhibition
CHANGES
IN SERUM SOLUBLE INTERLEUKIN 2 RECEPTOR LEVELS WITH CHRONIC HEPATITIS C BY INTERFERON-a. T. Takeda. S. Nishiwchi. S. Nakaiima. S. Shiomi. T. Kuroki. K, Third Department of Internal Medicine, Osaka City Kobavashi. University Medical School, Osaka, Japan.
IN PATIENTS
known
hepatitis.
Serum soluble interleukin to increase in patients
We
examined
the
2 receptor
(sIL2R)
levels
with acute and chronic relationship between sIL2R
are
viral and
eff’icaqy of interferon (IFN) treatment for patients with chronic hepatitis C (CH-C). We examined the time course changes in sIL2R in sera of patients during and 12 months after treatment. Human lymphoblastoid IFN was injected (im) at a dose of 6 Mega U/day every day for 2 weeks, and three times per week for 10 to 22 weeks. Serum sIL2R levels were measured using an enzyme immunoassay. The ratio of serum sIL2R at each time point Lo that before treatment was used to examine changes in sIL2R levels.
Fifty-nine
patients
with
CH-C
were
studied.
They
included
29
patients whose serum continued to be positive for HCVRNA after treatment (NR group). And 30 patients in whom HCVRNA was eliminated from serum and continued 10 be negative (CR group). In the patients, slL2R levels were correlated with ALT levels (r=O.465, p
1995
QUANTITATIVE ANALYSIS OF INTERFERON o / p RECEPTOR mRNA IN THE LIVER OF PATIENTS WITH CHRONIC HEPATITIS C H.Yatsuhashi,MC.Parauet.K.Yama&i,F.Iio.T.Aritomi.M.Yamasaki, O.Inoue.M.Kopa.M.Yano. Institute for Clinical Research, Nagasaki Chuo National Hospital, Nagasaki ,Japan.
association
by the
and response
hepatitis C v$us.
919
October
Background and methods:IFN a /,l3 receptor identified by Novick (1994) binds and responds effectively to IFN /3 and to several IFN (Y subtypes. But there has been no report on IFN (Y/P receptor associated with liver disease.We used RT-PCR method for the relative quantification of gene expression using a simultaneously amplified sequence of /3-a& mRNA as a internal control for the target sequence of IFN a /,L? receptor mRNA in the liver of 29 patients with chronic hepatitis C and in 6 controls with fatty liver.The PCR product of the internal control was reduced by delaying the addition of the primers for p-a&n. The cycle differences between the logarithmic phase of the curves for target sequence and for the p actin(&ycle) measured with a densitometry indicated the level of IFN (I /,8 receptor mRNA. Results:IFN a / p receptor mRNA in all patients and conlrols were detected. In chronic hepatitis C the mean concentration of IFN a / ,8 receptor mRNA was 2.0 fold higher than in the controls. There was no correlation between mRNA levels and ALT levels or histological findings in chronic hepatitis C. There was a significant reverse correlation between mRNA levels and serum HCV-RNA levels measured by b-DNA probe assay.(r=0.5589 P
Results: mRNA transcripts for IL-Q3 @=0.00079), IL-6 (p=O.O0017), IL-8 (p=O.O017), and TNFa @=0.00014) were detected in low numbers in 24
impairment
HEPATOLOGY
ABSTRACTS
920
with
biological
activity
of IFN
for IFN treatment in chronic
to reduce
viral replication
hepatitis C .
TRANSFORMING GROWTH FACTOR431 (TGFM) IN CHRONIC HEPATITIS C Nelson DR. Marousis CG. Oian RP. Xu Y. Gonzalez-Per&a RP. Davis CL, Lau JYN. Section of Hepatobiiary Diseases, University of Florida, Gainesville, FL. Baclmrouod: TGFal has been implicated in mediating hepatic tibrogenesis. TGP t31 is also known to have. negative regulatory effects on the immune system. Serum Th2 cytokines are elevated in chronic HCV infection. Hvuothesis: TGF-Bl production is increased ia chronic HCV infection. Sweitic Alms: (1) To determine the serum levels of TGFDl (total and biologically active) in chronic HCV infection and their clinical implications. (2) To study the relationship of TGFal with Th2 cytokiues (IL4, IL-10).&&&z Serum samples were prospectively collected on 88 chronic HCV patients (M:F 49:39, age 23-68) and 22 healthy controls (M:F U:9 age 19-63). Total (using acid activation) and biologically active TGFa 1, IL4, and IL10 were measured by ELISA. HCV RNA levels were quantitated by bDNA (Chiron) and genotypes were determined by RFLP based oa SUTR. Histological diagnosis was available in.87 patients (54 sections were available for complete analysis for disease activity). Liver sections from 80 other HCV patients were evaluated for hepatic expression of TGFal using immuaohistochemistry. m: HCV patients had a higher level of TGFal, both total (817f 464 rig/ml) and the biologically active form (52Or 370 pg/ml, 0=88), compared to controls (n=U, total TGFal: 183i 105 w/ml. n