Serum complement C4a and its relation to liver fibrosis in children with chronic hepatitis C.

AIM: To evaluate serum complement C4a and its relation to liver fibrosis in children with chronic hepatitis C virus (HCV) infection. METHODS: The study included 30 children with chronic HCV infection before receiving antiviral therapy. Chronic HCV infection was defined by positive anti-HCV, a positive polymerase chain reaction for HCV-RNA for more than 6 mo with absence of any associated liver disease. A second group of 30 age- and sex-matched healthy children served as controls. Serum C4a levels were measured by enzyme-linked immunosorbent assay. Liver fibrosis stage and inflammatory grade were assessed using Ishak scoring system. Serum C4a levels were compared according to different clinical, laboratory and histopathological parameters. Statistical significance for quantitative data was tested by Mann-Whitney U non-parametric tests. For qualitative data, significance between groups was tested by χ(2) test. Correlation was tested by Spearman's test. Results were considered significant if P value ≤ 0.05. RESULTS: The age of the patients ranged from 3.5 to 18 years and that of controls ranged from 4 to 17 years. C4a mean levels were merely lower in patients (153.67 ± 18.69 mg/L) than that in the controls (157.25 ± 11.40 mg/L) with no statistical significance (P = 0.378). It did not differ significantly in patients with elevated vs those with normal transaminases (152.25 ± 16.62 vs 155.36 ± 21.33; P = 0.868) or with different HCV viremia (P = 0.561). Furthermore, there was no statistical significant difference in serum levels between those with no/mild fibrosis and those with moderate fibrosis (154.65 ± 20.59 vs 152.97 ± 17.72; P = 0.786) or minimal and mild activity (155.1 ± 21.93 vs 152.99 ± 17.43; P = 0.809). Though statistically not significant, C4a was highest in fibrosis score 0 (F0), decreasing in F1 and F2 to be the lowest in F3. When comparing significant fibrosis (Ishak score ≥ 3) vs other stages, C4a was significantly lower in F3 compared to other fibrosis scores (143.55 ± 2.33 mg/L vs 155.26 ± 19.64 mg/L; P = 0.047) and at a cutoff value of less than 144.01 mg/L, C4a could discriminate F3 with 76.9% sensitivity and 75% specificity from other stages of fibrosis. CONCLUSION: Serum complement C4a did not correlate with any of transaminases, HCV viremia or with the histopathological scores. Although C4a decreased with higher stages of fibrosis, this change was not significant enough to predict individual stages of fibrosis. Yet, it could predict significant fibrosis with acceptable clinical performance.

Diagnostic value of hepatic intercellular adhesion molecule-1 expression in Egyptian infants with biliary atresia and other forms of neonatal cholestasis.

AIM: The diagnosis of biliary atresia (BA) is challenging as no single preoperative test is 100% accurate, especially for distinguishing it from other causes of neonatal cholestasis (NC). Intercellular adhesion molecule (ICAM) elevation was reported in BA as a part of the immune-mediated inflammatory process. The use of ICAM-1 as a discriminative tool between BA and other causes of NC has never been addressed before. This study was to evaluate the diagnostic potentials of ICAM-1 in BA versus other forms of NC. METHODS: For this purpose, serum ICAM-1 (sICAM-1) and ICAM-1 expression, in liver biopsy using immunohistochemistry, were estimated in 30 patients with BA and compared to that in 20 patients with other forms of NC. sICAM-1 levels were compared to that in 20 healthy controls. RESULTS: sICAM-1 levels were significantly higher in BA (1055.9 ± 230.2 ng/mL) than that in cholestasis (604.8 ± 194.8 ng/mL) and the control groups (158.9 ± 78.7 ng/mL) (P < 0.0001). A cut-off value of 793.8 ng/mL had 86.7% sensitivity and 95% specificity in discriminating the BA from the cholestasis group. The biliary expression score of ICAM-1 at a cut-off value of 110 could discriminate between BA and other causes of NC with 100% sensitivity and specificity. Neither serum levels nor liver expression of ICAM-1 scores correlated with disease severity or with fibrosis stage. CONCLUSION: These results suggest that ICAM-1 has a diagnostic value in patients with BA and would be a promising helpful tool when investigating patients with NC.

Obstructive jaundice promotes intestinal-barrier dysfunction and bacterial translocation: experimental study.

BACKGROUND: Although clinical and experimental studies have demonstrated a correlation between obstructive jaundice and the development of sepsis, the mechanism has not been fully elucidated. PURPOSE: The aim of this study was to investigate the influence of biliary obstruction on bacterial translocation as a possible source of infection in cases of obstructive jaundice. MATERIAL AND METHODS: Two groups of 12 Wistar rats were examined: rats subjected to common bile duct (CBD) ligation (group A) and rats subjected to a sham operation (group B). After 7 days, blood samples were taken and liver, spleen, and mesenteric lymph nodes (MLNs) from the ileocecal area were removed, divided into small pieces, and cultured. Quantitative culture results were determined by the number of colony-forming units (CFU) per milliliter of homogenate. Bacterial translocation was defined as the presence of a positive culture of MLNs, blood, liver, and/or spleen. Samples for histopathological examination were taken from the mucosa of the ileum and the colon and evaluated for inflammatory and destructive changes. RESULTS: There was no evidence of bacterial translocation to MLNs, blood, spleen, or liver detected in any of the 12 sham-operated control rats. In contrast, bacterial translocation was demonstrated in 8 of the 12 CBD-ligated rats (P < 0.01). In all eight cases in which translocation occurred, Escherichia coli were cultured from the MLNs. There were no histological changes in the mucosal samples of the control animals. In the CBD-ligated rats, hyperemia, vacuolization, reduction of goblet cells, decreased mitotic activity, and infiltration by lymphocytes and polymorphonuclear leukocytes (PMNLs) were detected. Cases in which translocation occurred were significantly associated with decreased mitotic activity in the colon (r = -0.5, P < 0.01) and higher infiltration by PMNLs in the ileum (r = -0.62, P < 0.05). CONCLUSION: Obstructive jaundice in a rat model predisposes to bacterial translocation. This suggests a mechanism whereby jaundiced patients are susceptible to septic complication.

Autoantibodies in chronic liver disease.

Some hepatotropic viruses (HBV and HCV) are capable of triggering autoimmune phenomena and manifest the features of autoimmune hepatitis (AIH) in the course of the disease. Careful attention is required to differentiate between AIH and chronic viral hepatitis (CVH) before the selection of treatment. This study was performed to assess the prevalence of rheumatoid factor (RF), antinuclear antibodies (ANA), anti-smooth muscle antibodies (ASMA), anti-mitochondrial antibodies (AMA), anti-parietal cell antibodies (APCA), anti-liver/kidney microsomal antibodies type I (ALKMA1) and anti-neutrophil cytoplasmic antibodies (ANCA) among patients with chronic liver diseases (CLD), and to assess the diagnostic value of these autoantibodies and their relation to HCV viral load and genotype and treatment with interferon-alpha (IFN-alpha). Five groups of patients with CLD (HCV, HBV, HCC, AIH and schistosomal hepatic fibrosis {SHF}) as well as a group of age- and gender-matched healthy controls were enrolled in the study. All the studied persons were subjected to full clinical assessment and laboratory investigations, including liver function tests, hepatitis markers, and HCV RNA by PCR. Detection of ANA, ASMA, APCA, AMA and ALKMA-1 was done by indirect immunofluorescence technique, while ANCA and RF were detected by EIA and latex agglutination test respectively. Results showed a significantly higher prevalence of RF, ASMA and ANCA among patients with CHC, RF and ASMA among HCC patients and ASMA and ALKMA1, among AIH patients as compared to the control group. Patients with HBV and those with SHF had a non-significantly higher prevalence of RF, ASMA and ANCA compared to controls. However, AMA was not detected in this study, and APCA showed no significant difference between the studied groups. The occurrence of these autoantibodies was not significantly related to HCV viral load, HCV genotype or treatment with IFN-alpha. There was a significant association between the occurrence of RF, ANA, ASMA, and ALKMA1 and high ALT levels, and between the occurrence of ANA, ASMA and ALKMA-1 and high AST and ALP levels. In conclusion, autoantibodies are commonly found among patients with HCV infection. The co-existence of HCV infection and autoimmune hepatitis should be considered in patients who are positive for both viral markers and autoantibodies and thorough evaluation of patients must be performed before selection of treatment. Testing for RF, ASMA and ANCA may have a good diagnostic value, however, AMA is the least useful in diagnosis.

Circulating anti-HLA antibodies in patients with chronic hepatitis C: relation to disease activity.

The human leukocyte antigens (HLA) may influence host immune to infection. In the mean time chronic hepatitis C (CHC) results in the appearance of a variety of autoantibodies. We investigated the frequency of circulating anti-HLA antibodies and none organ specific autoantibodies in patients with chronic hepatitis C at different stages of disease activity. Sixty-seven untreated male patients with CHC (anti-HCV antibody and HCV RNA positive), in whom 38 had elevated serum alanine aminotransferase (ALT) levels and 29 persistently normal serum ALT values, and 23 age-matched normal male subjects were studied. None of them had a history of blood transfusion. Sera were analyzed for immunoglobulin G-anti-HLA class I and class II antibodies by enzyme-linked immunosorbant assay, and for non-organ-specific autoantibodies (antinuclear, anti-smooth muscle, anti-mitochondrial and anti-liver/kidney microsomes type 1 antibodies) using indirect immunofluorescence technique. Circulating anti-HLA class I and class II antibodies were detected in 15/67 (22.4 %) and 11/67 (16.4 %) respectively, while none of normal controls had detectable anti-HLA antibodies in the serum. The frequency of detecting anti-HLA antibodies was significantly higher in patients with elevated serum ALT than persistently normal serum ALT values (31.6 % vs 10.3 %; P = 0.039) and was associated with non-organ-specific serum autoantibodies in 11/15 (73.3 %) patients. Those with circulating anti-HLA antibodies had significantly higher levels of serum aminotransferases, gamma-glutamyl transpeptidase, viral load and necroinflammatory and fibrosis scores in liver biopsies than patients with negative anti-HLA antibody (P < 0.001). In conclusions, the presence of circulating antibodies against HLA class I and class II molecules in HCV antibodies may represent an autoimmune response to HLA antigens and may play a pathogenetic role in the induction of the HCV-related chronic liver disease.