Interleukin-2 (IL-2) expression in livers of patients with chronic hepatitis C virus (HCV) infection.

The studies performed till now have pointed to an increased serum levels of interleukin 2 (IL-2) in infection with hepatitis C virus (HCV). The present study was aimed at examining intrahepatic expression of IL-2 in children (n=15) and in adults (n=11) with chronic hepatitis C as well as its correlations with histological lesions and selected clinical data. The immunocytochemical techniques and in situ hybridization method were applied at light and electron microscopy level. Under the light microscope, expression of IL-2 was analysed semiquantitatively. As compared to the control material, in livers of both groups of chronic hepatitis C patients augmented expression of IL-2 was demonstrated. The reaction product was localized mainly in the cytoplasm of hepatocytes which was confirmed by hybridocytochemistry. The mean proportion of cells with positive reaction for IL-2 mRNA was significantly lower than the proportion of cells positive for the respective protein. No correlation was disclosed between IL-2 expression on one hand and grading or staging, alanine aminotransferase (ALT) and HCV RNA levels in serum on the other. At the ultrastructural level, IL-2 in hepatocytes was present mainly in the endoplasmic reticulum and mitochondria. Our studies have confirmed augmented expression of IL-2 in livers of patients with chronic hepatitis C and have demonstrated that hepatocytes represent the principal source of the cytokine in HCV in vivo infection. Moreover, expression of IL-2 in the infection was examined for the first time at the ultrastructural level. Mitochondrial localization of IL-2 suggests a direct involvement of the cytokine in disturbed function of the organelles

p53 immunocytochemistry and TP53 gene mutations in patients with chronic hepatitis C virus (HCV) infection.

Chronic infection with hepatitis C virus (HCV) is regarded as a risk factor for hepatocellular carcinoma (HCC), mostly in patients with liver cirrhosis. Present study aimed at evaluation of cellular expression of p53 protein, genetic TP53 changes in liver samples and anti-p53 in serum of patients with chronic hepatitis C virus infection. The expression of p53 protein were analysed by immunocytochemistry in liver biopsies from adult patients with chronic, long-lasting hepatitis C. In order to detect TP53 mutations, PCR/SSCP and sequencing were performed. Antibodies against p53 in serum were determined using enzyme immunoassay (ELISA).In two out of 14 examined patients TP53 point mutations were detected in the liver samples. In the first patient, a substitution of C to T was demonstrated in position 1 of the codon 250, resulting in substitution of proline by serine. The other patient carried a substitution of C to G in position 13274 of the intron 6. The patient carrying mutation in the codon 250 demonstrated morphological traits of liver cirrhosis and had high number of p53-immunoreactive cell nuclei in tissue. None of the patients manifested elevated titres of serum anti-p53. In the liver, significant positive correlations were disclosed between expression of p53 on one hand and grading and staging on the other. A negative correlation was disclosed between cellular expression of p53 and duration time of infection. In conclusions, genetic changes in TP53 can be detected also in non-neoplastic lesions linked to chronic HCV infection

Intracellular expression of the proliferative marker Ki-67 and viral proteins (NS3, NS5A and C) in chronic, long lasting hepatitis C virus (HCV) infection.

Hepatitis C virus (HCV) continues to represent the main causative agent of the hepatitis, which leads to chronic transformation of the process in 60-80% individuals. It remains unclear how far cellular expression of HCV proteins in vivo may represent an index of progression of the disease and of proliferative activity in the liver in chronic hepatitis C. Aim of the studies included detection and subcellular localization of three HCV proteins (NS3, NS5A and C) in liver biopsies from adults (n=19) with chronic, long lasting hepatitis C as related to hepatocyte proliferative activity. The immunocytochemical ABC (avidin biotin-peroxidase complex) technique was applied, alone or associated with the ImmunoMax technique. Results of the immunocytochemical tests were compared to histological alterations in liver biopsies, proliferation index and with selected clinical data. A significantly higher expression of NS3 protein was noted, as compared to expressions of NS5A and C proteins. In all the patients, cytoplasmic localization of all proteins dominated over nuclear localization (p0.05). At the level of electron microscopy, protein localization in endoplasmic reticulum (ER) membranes, mitochondria, perinuclear region and/or in hepatocyte cell nucleus was observed. No direct relationships could be demonstrated between expressions of HCV proteins and of Ki-67 antigen. No correlations could also be demonstrated between cellular expression of any HCV protein on one hand and grading or staging, alanine transaminase (ALT), serum level of HCV RNA or alpha-fetoprotein (AFP) on the other. However, positive correlations were disclosed between proliferative activity of hepatocytes on one hand and patient's age, grading and staging on the other. Advanced hepatic fibrosis correlated also with serum levels of AFP. The studies were supplemented with data on subcellular localization of HCV proteins. Moreover, they indicated that in HCV infection grading and staging, proliferative activity of hepatocytes and serum AFP level represent more valuable indices of the disease progress than those provided by cellular expression of three potentially oncogenic HCV proteins in vivo