Proposal of a predictive model for advanced fibrosis containing Wisteria floribunda agglutinin‐positive Mac‐2‐binding protein in chronic hepatitis C

Aim

We aimed to construct a predictive model for advanced fibrosis containing Wisteria floribunda agglutinin‐positive Mac‐2‐binding protein (WFA⁺‐M2BP) level in patients with chronic hepatitis C (CHC) and to validate its accuracy in an independent cohort.

Methods

A total of 386 patients with CHC were retrospectively analyzed. For the purpose of this study, we formed a training set (n = 210) and a validation set (n = 176). In the training set, we investigated variables linked to the presence of advanced fibrosis using univariate and multivariate analyses. We constructed a formula for predicting advanced fibrosis and validated its accuracy in the validation cohort. Receiver operating characteristic curve (ROC) analysis was carried out for calculating the area under the ROC (AUROC).

Results

In multivariate analyses, WFA⁺‐M2BP (P = 0.029) and prothrombin time (PT) (P = 0.018) were found to be significant predictive factors linked to the presence of advanced fibrosis; platelet count (P = 0.098) and hyaluronic acid (P = 0.078) showed borderline statistical significance for the presence of advanced fibrosis. Using these four variables (with the initials MPPH), we constructed the following formula: MPPH score = −3.584 − (0.275 × WFA⁺‐M2BP) + (0.068 × platelet count) + (0.042 × PT) − (0.005 × hyaluronic acid). In the training and validation sets, MPPH score yielded the highest AUROCs (0.87 and 0.83) for predicting advanced fibrosis among eight serum liver fibrosis markers. Similarly, in the training and validation sets, MPPH score had the highest diagnostic accuracies for predicting advanced fibrosis among eight serum variables (81.4% and 74.4%).

Conclusion

Our proposed MPPH scoring system can be useful for predicting advanced fibrosis in patients with CHC.

     

Serum Wisteria floribunda agglutinin-positive Mac-2-binding protein expression predicts disease severity in chronic hepatitis C patients

Wisteria floribunda agglutinin-positive Mac-2 binding protein (WFA⁺-M2BP) has recently been developed as a promising liver fibrosis glyco biomarker. We assessed its efficacy in evaluating liver disease severity in chronic hepatitis C (CHC) in Taiwan. The association between WFA⁺-M2BP and histological features was evaluated among those CHC patients underwent liver biopsy. We also aimed to clarify the factors determining the performance of WFA⁺-M2BP in CHC. A total of 229 CHC patients were consecutively recruited. The mean value of WFA⁺-M2BP in patients from F0 to F4 was 1.68, 2.23, 3.45, 3.48, 3.77 respectively (linear trend P = 0.008). Linear regression analysis revealed that alanine aminotransferase (odds ratio [OR]: 0.03, 95% confidence intervals [CI]: 0.02–0.05, P < 0.001), AST (OR: ?0.1, 95% CI: ?0.02 to ?0.01, P < 0.001), and liver fibrosis (OR: 0.30, 95% CI: 0.01–0.59, P = 0.043) were the independent factors correlated to serum WFA⁺-M2BP level. The optimal cutoff values of WFA⁺-M2BP for fibrosis stages F1, F2, F3, and F4 were 1.42, 1.61, 1.42, and 2.67, respectively. Multivariate analysis revealed that the platelet count (OR/CI: ?0.009/0.986–0.996, P = <0.001), r-glutamyl transferase (OR/CI: 0.007/1.000–1.013, P = 0.036), and WFA⁺-M2BP (OR/CI: 0.187/1.057–1.374, P = 0.005). We concluded that WFA⁺-M2BP is a competent noninvasive marker for liver fibrosis assessment in CHC patients.     

Clinical significance of serum Wisteria floribunda agglutinin-positive Mac-2 binding protein in pancreatic ductal adenocarcinoma

BackgroundWisteria floribunda agglutinin-positive mac-2 binding protein (WFA⁺-M2BP) is an excellent biomarker for predicting hepatic fibrosis. We hypothesized WFA⁺-M2BP might be a serum biomarker for the diagnosis of chronic pancreatitis (CP) and pancreatic ductal adenocarcinoma (PDAC) with dense fibrosis.MethodsIn this study, we included 16 CP and 24 PDAC patients. Serum levels of WFA⁺-M2BP (cut-off index [COI]) were compared between the 2 groups. To confirm the cellular production of WFA⁺-M2BP, we investigated the presence of WFA⁺-M2BP in HEK293 cells, 3 established human PDAC cell lines and a recently generated human PDAC cell line derived from a liver metastasis (MDA-PATC53). The bio-physiological effects of MDA-PATC53 supernatant were evaluated. Finally, the difference in the expression of glycosylation enzymes between MDA-PATC53 and Panc-1 were analyzed by cDNA microarray.ResultsWe found that the serum WFA⁺-M2BP level could distinguish the 2 groups. The median serum COI of WFA⁺-M2BP was 0.98 and 0.51 in PDAC and CP, respectively. Additionally, WFA⁺-M2BP positive PDACs were more frequently associated with metastatic lesions than the WFA⁺-M2BP negative PDACs (91.6% vs. 41.7%, P = 0.009). The MDA-PATC53 cells alone produced WFA⁺-M2BP. However, we found that MDA-PATC53 supernatant containing WFA⁺-M2BP (1.0 COI) did not alter the biological behavior of cancer cell lines. The results of cDNA microarray revealed that several glycosylation enzymes with pro-oncologic function were highly expressed in MDA-PATC53 compared to Panc-1.ConclusionsSerum WFA⁺-M2BP can be a useful biomarker for the diagnosis of PDAC and the prediction of disease progression since it potentially reflects altered pro-oncologic glycosylation enzymes.     

Case report of diet‐related improvements of non‐alcoholic steatohepatitis evaluated by four consecutive liver biopsies

We report the case of a 46‐year‐old man who achieved remarkable improvement of characteristic non‐alcoholic steatohepatitis by dieting. Initially, his body mass index was 40.7 kg/m². He proactively commenced a low calorie diet and his body mass index decreased to 28 kg/m² after 4 years. During the same period, we carried out liver biopsy four times. Liver fibrosis as well as inflammation, steatosis and ballooning improved, and the non‐alcoholic fatty liver disease activity score decreased from 4 to 0. The fat deposits of liver tissue changed from macrovesicular droplets to microvesicular, and finally disappeared. Along with improved histology, alanine aminotransferase, homeostasis model assessment‐insulin resistance, ferritin, leptin, high‐sensitivity C‐reactive protein and cytokeratin 18 decreased, and adiponectin increased linearly. However, no clear changes were observed in markers for Wisteria floribunda agglutinin‐positive Mac‐2 binding protein (WFA⁺M2BP), type IV collagen 7S, procollage III peptide and fibrosis‐4 index along with dieting. This is the first study to report gradual improvement of liver histology during four liver biopsies and fibrosis markers in a patient with non‐alcoholic steatohepatitis who achieved complete recovery solely by dieting.     

Altered serum N‐glycomics in chronic hepatitis B patients

Background: We previously reported on serum N‐glycans as markers for the diagnosis of cirrhosis in patients with chronic hepatitis C infection. Our present study aimed to evaluate the use of serum glycan markers for the diagnosis of liver fibrosis in patients with chronic hepatitis B infection. Methods: Patients with hepatitis B virus (HBV) infection (n=173) were diagnosed by clinical laboratory analysis and histological examination. Liver fibrosis was staged using Ishak score. N‐glycan profiles of serum proteins were determined by DNA sequencer‐based carbohydrate analytical profiling. Results: We found that in HBV patients, like in hepatitis C virus patients, several serum N‐glycans were altered during the development of liver fibrosis. We found higher levels of total agalactosylated biantennary glycans in fibrosis patients with HBV infection than in healthy controls. The biantennary (NA2) and the triantennary (NA3) N‐glycans decreased significantly (P<0.001) with increased severity of fibrosis. The diagnostic power of serum glycan marker (GlycoFibroTest) [area under the curve (AUC)=0.735) was similar to that of FibroTest (AUC=0.740) for discriminating between moderate and advanced fibrosis (F3–F6) from non‐ or mild fibrosis (F0–F2). However, GlycoFibroTest (AUC=0.740) was slightly better than FibroTest (AUC=0.696) for distinguishing fibrotic patients (F1 or more) from non‐fibrotic patients (F0). Conclusions: The assay for serum glycan profiling showed satisfactory reproducibility and is a non‐invasive blood test for the diagnosis of liver fibrosis. The changes of N‐glycan level in serum can be used to monitor or follow‐up the progress of fibrosis using specific N‐glycan markers.     

Hepatitis C viraemia reversibly maintains subset of antigen‐specific T‐bet+ tissue‐like memory B cells

Background: Chronic antigen exposure and/or ageing increases the frequency of T‐box expressed in T cells (T‐bet)‐expressing B‐lymphocytes in mice. The frequency and significance of B‐cell T‐bet expression during chronic hepatitis C (HCV) infection in human subjects has never been described. Methods: Healthy controls, cirrhotic and noncirrhotic HCV‐infected patients, and non‐HCV patients with cirrhosis were recruited. Peripheral blood mononuclear cells were phenotyped for expression of T‐bet and related markers by flow cytometry. In a subset of patients who underwent antiviral therapy and were cured of HCV infection (sustained virological response), the dynamics of T‐bet expression in B cells was monitored. After cure, convalescent B cells were tested for T‐bet expression after re‐exposure to infected plasma or recombinant HCV proteins. Results: Forty‐nine patients including 11 healthy donors, 30 hepatitis C‐infected individuals (nine with liver cancer, 13 with cirrhosis, eight without cirrhosis) and eight patients with cirrhosis due to non‐HCV‐related cause were recruited. We found that B cells in patients with chronic HCV exhibited increased frequency of T‐bet+ B cells relative to noninfected individuals (median 11.5% v. 2.2%, P<.0001) but that there were no significant differences between noncirrhotic, cirrhotic and cancer‐bearing infected individuals. T‐Bet⁺ B cells expressed higher levels of CD95, CXCR3, CD11c, CD267 and FcRL5 compared to T‐bet^? B cells and predominantly exhibit a tissue‐like memory CD27^?CD21^? phenotype independent of HCV infection. T‐bet⁺ B cells in HCV‐infected patients were more frequently class‐switched IgD^?IgG⁺ (40.4% vs. 26.4%, P=.012). Resolution of HCV infection with direct‐acting antiviral (DAA) therapy leads to a marked reduction in the frequency of T‐bet⁺ B cells (median 14.1% pretreatment v. 6.7% end of treatment v. 6.1% SVR12, P≤.01). Re‐exposure of convalescent (cured) B cells to viremic plasma and recombinant HCV E2 protein led to re‐expression of T‐bet. Conclusion: Chronic antigenemia in chronic HCV infection induces and maintains an antigen‐specific T‐bet⁺ B cell. These B cells share markers with tissue‐like memory B cells. Antigen‐driven T‐bet expression may be a critical suppressor of B‐cell activation in chronic HCV infection.     

Serial measurement of <Emphasis Type="Italic">Wisteria floribunda agglutinin</Emphasis> positive Mac-2-binding protein is useful for predicting liver fibrosis and the development of hepatocellular carcinoma in chronic hepatitis C patients treated with IFN-based and IFN-free therapy

Aim

Wisteria floribunda agglutinin positive (WFA⁺) Mac-2-binding protein (M2BPGi) is a noninvasive glyco-marker for liver fibrosis. This study evaluated the utility of serial measurement of serum M2BPGi and total M2BP as a predictor of fibrosis and the development of hepatocellular carcinoma (HCC).

Methods

This study included 119 patients with chronic hepatitis C (CHC). Of these patients, 97 were treated with IFN-based therapy and 22 were treated with daclatasvir and asunaprevir. Serum M2BPGi values were measured prior to, at the end of, and at 24 weeks after the completion of treatment. As subanalysis, serum total M2BP levels were measured in patients treated with pegylated-interferon and ribavirin.

Results

In patients treated with IFN-based therapy, M2BPGi levels were elevated at the end of treatment but decreased afterwards. In contrast, M2BPGi levels in patients treated with IFN-free therapy decreased immediately after starting the treatment without transient elevation. Though pre-treatment M2BPGi levels significantly correlated with fibrosis in both patients with a sustained virological response (SVR) and non-SVR, post-treatment M2BPGi levels decreased regardless of the degree of fibrosis in patients with SVR. In multivariate analysis, non-SVR and HCC development were independent factors associated with M2BPGi level ≥2.2. In patients treated with pegylated-interferon and ribavirin, total M2BP levels were positively correlated with fibrosis and HCC development.

Conclusion

Real-time monitoring of the serum M2BPGi level after antiviral therapy for CHC patients could be a helpful screening tool for assessing the risk of HCC. M2BP and its glycan structure could be associated together with hepatocarcinogenesis.

     

Liver‐infiltrating CD56 positive T lymphocytes in hepatitis C virus infection

Abstract: Aim: Hepatitis C virus (HCV) is a major cause of post‐transfusional and sporadic hepatitis, and leads to chronic liver disease. It has been suggested that virus‐specific cytotoxic T lymphocytes are responsible for liver injuries that occur in HCV‐infected patients. However, the detailed characteristics of these lymphocytes have not yet been defined. We have previously reported that CD56⁺ T lymphocytes, as intermediates between natural killer cell and T lymphocytes, predominantly infiltrated the liver and were increased in patients with chronic hepatitis related to HCV (CH‐C). Material and Methods: We obtained peripheral blood and liver tissues from 32 patients diagnosed as having CH‐C, and 10 other liver disease patients (5 chronic hepatitis related to HBV, 5 alcoholics), and analyzed peripheral blood and liver‐infiltrating lymphocytes using flow cytometric and immunohistochemical techniques. Results: The CD56⁺ T lymphocyte ratio in the liver of patients with a high histology activity index (HAI) score for chronic hepatitis was higher than that of patients with a low HAI score and patients with other liver diseases. In addition, T lymphocytes from patients with chronic hepatitis with a high HAI score carried mostly γδ‐TCR. There was a correlation between the ratio of CH‐C and serum alanine aminotransferase, category I (periportal inflammation and necrosis), and IV (fibrosis) of the HAI scoring system. The ratio was highest in zone 1 of the hepatic lobules. Conclusion: The correlation between CD56⁺ T lymphocyte ratios and hepatocellular damage was examined. These findings suggest strongly that liver‐infiltrating CD56⁺ T lymphocytes play an important pathologic role in hepatocellular injury in CH‐C.     

Compartmentalization and its implication for peripheral immunologically‐competent cells to the liver in patients with HBV‐related acute‐on‐chronic liver failure

Aims: This study attempts to characterize the feature of immunologically competent cells (ICCs) and evaluate its clinical implication in patients with acute‐on‐chronic liver failure (ACLF) in relation to chronic hepatitis B virus (HBV) infection. Methods: Circulating ICCs were examined in ACLF patients (n = 75), as well as in patients with hepatitis B (CHB, n = 31), CHB‐related liver cirrhosis (LC, n = 36), and normal controls (NC, n = 30). Intrahepatic ICCs in some patients were further analyzed via immunohistochemical and flow cytometric assays. Results: Total lymphocytes, CD4⁺ T cells, CD8⁺ T cells, and NK cells in circulation were numerically lower in the ACLF and LC groups compared to the CHB and NC groups. Importantly, the number of these cells was significantly lower in non‐surviving ACLF patients compared with surviving ACLF patients. In comparison to NC, ACLF patients displayed a significantly higher ratio of liver‐infiltrating CD4⁺ T‐cell frequency than its circulating counterpart, suggesting that the possiblility of the ICCs compartmentalization from the peripheral blood into the liver in ACLF. Immunohistochemical analysis showed that intrahepatic CD4⁺ cells, CD8⁺ cells, and CD56⁺ cells were significantly higher in the ACLF group compared with the other three groups, suggesting a stronger cellular immune response‐mediated inflammation in ACLF group than other patient groups. Conclusions: The abnormal prevalence of circulating and intrahepatic ICCs possibly acts as an important factor that may drive the progression of HBV‐related ACLF.     

Impact of oral silymarin on virus‐ and non‐virus‐specific T‐cell responses in chronic hepatitis C infection

Silymarin displays anti‐inflammatory effects on T lymphocytes in vitro. The immunomodulatory properties of oral silymarin in vivo in humans with chronic hepatitis C have not previously been characterized. We hypothesized that silymarin would suppress T‐cell proliferation and pro‐inflammatory cytokine production of virus‐ and non‐virus‐specific T cells while increasing anti‐inflammatory IL‐10 production in vivo. Patients from one site of the SyNCH‐HCV double‐masked, placebo‐controlled study of oral silymarin in prior interferon nonresponders with chronic hepatitis C provided blood samples at baseline and treatment week 20. Mononuclear cells were stimulated with recombinant HCV proteins and controls in ³H‐thymidine proliferation assays, IFNγ ELISPOT and IL‐10 ELISPOT. The frequency of CD4⁺CD25^hi and CD4⁺foxp3⁺ regulatory T cells, serum cytokine levels, serum IP‐10 and lymphocyte interferon‐stimulated gene expression were also quantified at baseline and week 20. Thirty‐two patients were recruited (10; placebo, 11; 420 mg three times a day, 11; 700 mg three times a day). Serum ALT and HCV RNA titres did not change in any group. HCV‐specific CD4⁺ T‐cell proliferation and the frequency of IFNγ‐ and IL‐10‐producing T cells were not significantly changed in silymarin‐treated subjects. However, C. albicans‐induced T‐cell IFNγ and phytohaemagglutinin‐induced T‐cell proliferation were suppressed by silymarin therapy. A trend towards augmentation of interferon‐induced ISG15 expression was present in the high‐dose silymarin group. While no effect on HCV‐specific T cells was identified, these data confirm that high‐dose oral silymarin exerts modest nonspecific immunomodulatory effects in vivo. The impact of this anti‐inflammatory effect on long‐term liver health in chronic hepatitis C merits future clinical investigation.     

Increased levels of IL‐21 responses are associated with the severity of liver injury in patients with chronic active hepatitis B

Interleukin‐21 (IL‐21) participates in tissue damage in various immune‐mediated diseases. Its role in the pathogenesis of chronic active hepatitis B (CAHB) has not been clarified. The frequency of circulating IL‐21⁺ T cells and the levels of serum and intrahepatic IL‐21 have been characterized in 70 CAHB patients, 32 inactive carrier (IC), 18 chronic hepatitis C (CHC) and 20 healthy controls (HC). Their potential association with liver injury was analysed. The percentages of IL‐21⁺CD3⁺CD8⁻ and IL‐21⁺CD3⁺CD8⁺ T cells and the levels of serum IL‐21 in CAHB patients were significantly higher than that in the IC, CHC patients and HC (P < 0.001) and were correlated positively with the levels of serum alanine aminotransferase (ALT, r = 0.424, P < 0.001; r = 0.392, P = 0.001) and aspartate aminotransferase (AST, r = 0.388, P = 0.001; r = 0.329, P = 0.005) in CAHB patients, respectively. The levels of IL‐21 expression in the liver tissues were associated significantly with increased degrees of inflammation and fibrosis in CAHB patients (P < 0.01 or P < 0.05). Our findings suggest that aberrant IL‐21 responses may be associated with the progression of CHB.     

The gamma‐glutamyl transpeptidase‐to‐albumin ratio predicts significant fibrosis and cirrhosis in chronic hepatitis B patients

Background/Aim Simple, inexpensive and clinically available noninvasive liver fibrosis tests are highly needed. We aimed to develop a novel noninvasive index for predicting significant fibrosis and cirrhosis in chronic hepatitis B (CHB) patients. Methods Using liver histology as gold standard, we developed a novel index to predict significant fibrosis and cirrhosis in CHB patients and then compared the diagnostic accuracy of the novel index, aspartate transaminase‐to‐platelet ratio index (APRI), and fibrosis index based on four factors (FIB‐4) in a training set (606 patients) and a validation set (216 patients) from the same patient catchment area. Results Of 606 CHB patients in the training set, 33.2% had significant fibrosis and 11.4% had cirrhosis. In multivariable analysis, gamma‐glutamyl transpeptidase (GGT) (OR=1.032, p<0.001) and albumin (OR=0.953, p=0.048) were independent predictors of significant fibrosis. Consequently, a GGT‐to‐albumin ratio (GAR) was developed. In the training set, the area under the receiver operating characteristic curve (AUROC) of GAR was significantly higher than that of APRI and FIB‐4 to predict ≥F2 (0.82, 0.70, and 0.68, respectively), ≥F3 (0.86, 0.76, and 0.75, respectively), and F4 (0.88, 0.75, and 0.73, respectively), respectively. In the validation set, the AUROC of GAR was also better than APRI and FIB‐4 for predicting ≥F2 (0.81, 0.63 and 0.61, respectively), ≥F3 (0.88, 0.78, and 0.76, respectively) and F4 (0.92, 0.85, and 0.78, respectively), respectively. Conclusions GAR is a more accurate noninvasive index than APRI and FIB‐4 to stage significant fibrosis and cirrhosis in CHB patients and represents a novel noninvasive alternative to liver biopsy.     

Possible involvement of chemokine C‐C receptor 7−programmed cell death‐1+ follicular helper T‐cell subset in the pathogenesis of autoimmune hepatitis

Background and Aim

Recent studies have demonstrated that B cells and follicular helper T (Tfh) cells, which are central regulators of humoral immune response, contribute to the development and progression of autoimmune diseases. Because Tfh cells can be divided into several subsets with distinct functional properties, this study aimed to examine the roles of different subsets of circulating Tfh cells in the immune pathogenesis of autoimmune hepatitis (AIH).

Methods

Thirty‐five patients with AIH, 28 patients with primary biliary cholangitis, 22 patients with chronic hepatitis B (CHB), and 44 health controls (HC) were enrolled. The frequencies of different Tfh subsets in the blood and liver were examined by flow cytometry and immunohistochemical staining. The function of circulating Tfh subsets was examined after in vitro stimulation.

Results

In newly diagnosed AIH patients, the frequency of circulating chemokine C‐C receptor 7^?programmed cell death‐1⁺ Tfh subset was significantly increased compared with that in CHB patients and HC, significantly correlated with clinical parameters, including serum IgG, prothrombin time and albumin levels, and significantly decreased after corticosteroid treatment. In the liver of AIH patients, the frequencies of activated Tfh subsets were significantly increased and positively correlated with those in the blood. Moreover, the ability to produce interleukin‐21 and interleukin‐17 from circulating Tfh cells was significantly increased in AIH patients compared with HC.

Conclusions

These results significantly extend our understanding of Tfh subsets in AIH and suggest a potential role of dysregulated chemokine C‐C receptor 7^?programmed cell death‐1⁺ Tfh subset in the pathogenesis and disease progression of AIH.     

Non‐invasive assessment of liver fibrosis by stiffness measurement in patients with chronic hepatitis B

Background: The need for new non‐invasive tools to assess liver fibrosis in chronic liver diseases has been largely advocated. Liver stiffness measurement (LSM) using transient elastography (FibroScan^®, Echosens^?) has been shown to be correlated to liver fibrosis in various chronic liver diseases. This study aims to assess its diagnosis accuracy in patients with chronic hepatitis B. Patients and methods: We prospectively enrolled 202 patients with chronic hepatitis B in a multicentre study. Patients underwent liver biopsy (LB) and LSM. METAVIR and Ishak liver fibrosis stages were assessed by two pathologists. Results: LSM or LB was considered unreliable in 29 patients. Statistical analysis was conducted in 173 patients. LSM was significantly (P<0.001) correlated with METAVIR (r=0.65) and Ishak fibrosis stage (0.65). The area under receiver‐operating characteristic curves were 0.81 (95% confidence intervals, 0.73–0.86) for F≥2, 0.93 (0.88–0.96) for F≥3 and 0.93 (0.82–0.98) for F=4. Optimal LSM cut‐off values were 7.2 and 11.0 kPa for F≥2 and F=4, respectively, by maximizing the sum D of sensitivity and specificity, and 7.2 and 18.2 kPa by maximizing the diagnosis accuracy. Conclusion: In conclusion, LSM appears to be reliable for detection of significant fibrosis or cirrhosis in HBV patients and cut‐off values are only slightly different from those observed in HCV patients.     

Circulating myeloid‐derived suppressor cells correlate with clinical outcome in Hodgkin Lymphoma patients treated up‐front with a risk‐adapted strategy

In the attempt to find a peripheral blood biological marker that could mirror the dysregulated microenvironment of Hodgkin Lymphoma (HL), we analysed the amount of myeloid‐derived suppressor cells (MDSC), including the three main sub‐types (monocytic, granulocytic and CD34 + fraction). The absolute MDSC count was investigated in 60 consecutive newly diagnosed HL patients and correlated with clinical variables at diagnosis and outcome. Patients received standard‐of‐care chemotherapy with the exception of interim fluorodeoxyglucose positron emission tomography (PET‐2)‐positive patients, who were switched early to a salvage regimen. All MDSC subsets were increased in HL patients compared to normal subjects (P < 0·0001) and were higher in non‐responders. However, a strong prognostic significance was limited to immature (CD34⁺) MDSC. A cut‐off level of 0·0045 × 10⁹/l for CD34⁺MDSC resulted in 89% (95% confidence interval [CI] 52–99%) sensitivity and 92% (95% CI 81–98%) specificity. The positive predictive value to predict progression‐free survival was 0·90 for PET‐2 and 0·98 for CD34⁺MDSC count; the negative predictive value was 0·57 for PET‐2 and 0·73 for CD34⁺MDSC. PFS was significantly shorter in patients with more than 0·0045 × 10⁹ CD34⁺MDSC cells/l at diagnosis and/or PET‐2 positivity (P < 0·0001). In conclusion, all circulating MDSC subsets are increased in HL; CD34⁺MDSC predict short PFS, similarly to PET‐2 but with the advantage of being available at diagnosis.     

A prospective study of T‐ and B‐lymphocyte subpopulations,CD81 expression levels on B cells and regulatory CD4+CD25+CD127low/−FoxP3+ T cells in patients with chronic HCV infection during pegylated interferon‐alpha2a plus ribavirin treatment

Summary. Resolution of hepatitis C virus (HCV) infection requires a complex interplay between innate and adaptative immune responses. The role of lymphocyte subpopulations during combined antiviral treatment remains to be defined. This study was conducted to assess the effect of pegylated interferon‐alpha2a (pegIFN‐α2a) and ribavirin treatment on peripheral blood lymphocytes, mainly on CD81 expression on B cells and CD4⁺CD25⁺CD127^low/?FoxP3⁺ regulatory T cells (Tregs) in patients with chronic HCV infection. Thirty‐five patients with chronic HCV infection who started pegIFN‐α2a and ribavirin treatment were enrolled. Peripheral blood mononuclear cells (PBMC) were obtained at baseline before treatment (BT), mid‐treatment (MT), the end of treatment (ET) and 24 weeks post‐treatment (PT). During combined antiviral treatment, a significant decrease in the percentage of CD3⁺, CD8⁺, CD3⁺gamma/delta (γδ)⁺, CD19⁺ lymphocyte subpopulations and Tregs was observed. There was also a significant increase in the percentage of the CD4⁺ lymphocyte subpopulation and in CD81 expression levels on CD19⁺ B cells when BT was compared with ET (all P < 0.05). Seventeen patients were nonresponders (NR) and 18 had a sustained virological response (SVR). At baseline, NR patients had higher CD81 expression levels on CD19⁺ B cells (P = 0.017) and a higher Tregs percentage (P = 0.025) than SVR patients. Our results suggest that immunomodulation fluctuates during antiviral treatment and that percentage CD81 expression levels on B cells and Tregs might be useful as an immunological prognostic factor for pegIFN‐α2a and ribavirin treatment response in chronic HCV infection.     

Clinical,virological and histopathological features: long‐term follow‐up in patients with chronic hepatitis C co‐infected with S. mansoni

Abstract: Background/Aims: Infection with Schistosoma mansoni is endemic in Egypt leading to hepatic schistosomiasis and eventually portal hypertension. The prevalence of antibodies against hepatitis virus C among Egyptians is 14–51%. The aim of the present study was to investigate the influence of schistosomiasis on chronic hepatitis C with respect to the natural course of the disease, immunology, virology and histology. Patients and Methods: One hundred and twenty‐six Egyptian patients classified into three groups: group A: chronic hepatitis C (n=33); group B: chronic schistosomiasis (n=30) and group C: chronic hepatitis C and chronic schistosomiasis (n=63) were enrolled and prospectively followed for 62.7±22 months. Patients infected with other hepatic viruses and/or parasites were excluded. Detailed history, clinical examination, CD4⁺ and CD8⁺ lymphocyte counts in blood, hematological and blood chemical values, abdominal ultrasonography, upper endoscopy, HCV RNA titer by RT/PCR, genotype and histological activity index in the liver biopsy were determined. Results: Thirty patients (48%) with HCV and schistosomiasis had liver cirrhosis and Child‐Pugh class C vs. five (15%) in HCV patients and none in the schistosomal group. HCV RNA levels ranged between 0.07 and 13×10⁵ copies/ml in group A, and between 1 and 25×10⁵ copies/ml in group C. HCV genotype 4 was detected in 58 patients with co‐infection (92%) and 21 patients with HCV alone (64%). Patients with coinfection showed higher grading and staging scores in their liver biopsies. Hepatocellular carcinoma was detected only in patients with coinfection. During follow‐up, the mortality rate was 12%, 3% and 48% in group A, B and C, respectively. Conclusions: Patients with concomitant HCV and schistosomiasis infection were characterized by more advanced liver disease, higher HCV RNA titers, predominance of HCV genotype 4, higher histologic activity, higher incidence of cirrhosis and hepatocellular carcinoma as well as a much higher mortality rate.     

Prevalence and clinical characterization of patients with acute hepatitis B induced by lamivudine‐resistant strains

Background and Aims: Acute hepatitis caused by lamivudine (LMV)‐resistant strains has not been reported, and the clinical impact of LMV‐resistant strains on acute hepatitis is not known. The aim of this study was to investigate the molecular and clinical characteristics of patients with acute hepatitis B caused by LMV‐resistant strains. Methods: Forty‐five patients with acute hepatitis B were studied. Hepatitis B virus (HBV) subgenotypes and LMV‐resistance mutations were determined by direct sequencing of the preS and polymerase regions, respectively. Results: HBV subgenotypes A2 (n = 18), B1 (n = 1), B2 (n = 3), B3 (n = 2), C1 (n = 1), C2 (n = 19) and C6 (n = 1) were detected in patients with acute hepatitis. LMV‐resistance mutations were detected in two patients. LMV‐resistance mutations (L180M, M204I) were detected in a patient with subgenotype C2 who had acute self‐limited hepatitis. The other patient with LMV‐resistance mutations (L180M, M204V) was infected with subgenotype A2 and had severe hepatitis. Conclusion: LMV‐resistant strains are rare, but they are starting to be found in patients with acute hepatitis B. Surveillance for detecting drug‐resistant HBV strains would be important for clinical practice.     

Evolution of fibrosis during HCV recurrence after liver transplantation – influence of IL‐28B SNP and response to peg‐IFN and ribavirin treatment

The IL‐28 gene is associated with sustained viral response (SVR) after treatment with peg‐IFN and ribavirin in liver transplant recipients with chronic hepatitis C genotype 1 infection. We analysed the importance of recipient and donor IL‐28B genotype for response to treatment and fibrosis progression in 54 liver transplant recipients. Fibrosis stage (F) was defined as mild when F ≤ 2 and severe when F ≥ 3 in a liver biopsy or according to liver elasticity analysis. We found a significantly lower prevalence of IL‐28B SNP CC in the recipients (22%) than in the donors (67%), P < 0.0001. SVR was seen in 61% of the recipients with mild and 27% with severe fibrosis pretreatment, P = 0.01. Recipients with IL‐28 CC and non‐CC had mild fibrosis in 64% and 38% prior to treatment, P = 0.13. At follow‐up, after treatment, significantly more recipients with CC had mild fibrosis than non‐CC recipients (75% versus 32%, P = 0.0072), and all with CC and SVR had mild fibrosis. The strongest baseline factor predicting SVR was genotype. Hence, 13/19 (68%) genotype non‐1 patients reached SVR versus only 9/35 (26%) genotype 1 patients, P = 0.0022. In summary, we found that liver transplant recipients with IL‐28B CC tended to have less advanced fibrosis prior to and significantly less after SOC treatment and that all recipients with IL‐28B CC who achieved SVR had mild fibrosis at follow‐up. A significantly higher SVR rate was achieved in recipients with mild than severe fibrosis pretreatment and with genotype non‐1 than 1 infection. Our findings indicate that treatment for post‐transplant HCV recurrence should be offered before advanced fibrosis is seen in the recipient.     

High rate of long‐term virological response after a 1‐year course of interferon ± ribavirin in chronic hepatitis C relapsers. Results of a 191 patients randomized trial

We investigated the long‐term efficacy of a 12‐month course of interferon (IFN)+ribavirin in chronic hepatitis C relapsers. We randomized 191 relapsers with a 2:1 ratio to receive 3 million units three times a week of interferon alpha (IFN α)‐2b+ribavirin (1–1.2 g/day) (group A=127 patients) or IFN α‐2b (group B=60 patients) of same dosage and duration for 1 year. General and hepatitis C data of group A and B patients were similar. The main goal of the study was to determine the rate of sustained virological response evaluated 1 year after treatment. Results: Virological sustained response (SR) was 61% and 12% for groups A and B, respectively (P<0.001). A significant histological improvement was observed in both treatment groups. The Metavir activity score became significantly lower in the IFN+ribavirin group than in the IFN group (P<0/0001). The Metavir fibrosis scores remained unchanged. Also, at the end of the treatment, the virological response was 69% (88/127) for group A and 33% (20/60) for group B (P<0.001). Conclusion: One‐year retreatment of relapsers with the combination of IFN+ribavirin led to 61% of virological SR and to a significant improvement of histological activity. Therefore, the therapeutic schedule presented here can be considered of particular interest for the retreatment of relapsers.