Chao-Shuang Lin

Amniotic-fluid-derived mesenchymal stem cells overexpressing interleukin-1 receptor antagonist improve fulminant hepatic failure.

Uncontrolled hepatic immunoactivation is regarded as the primary pathological mechanism of fulminant hepatic failure (FHF). The major acute-phase mediators associated with FHF, including IL-1β, IL-6, and TNF-α, impair the regeneration of liver cells and stem cell grafts. Amniotic-fluid–derived mesenchymal stem cells (AF-MSCs) have the capacity, under specific conditions, to differentiate into hepatocytes. Interleukin-1–receptor antagonist (IL-1Ra) plays an anti-inflammatory and anti-apoptotic role in acute and chronic inflammation, and has been used in many experimental and clinical applications. In the present study, we implanted IL-1Ra–expressing AF-MSCs into injured liver via the portal vein, using D-galactosamine–induced FHF in a rat model. IL-1Ra expression, hepatic injury, liver regeneration, cytokines (IL-1β, IL-6), and animal survival were assessed after cell transplantation. Our results showed that AF-MSCs over-expressing IL-1Ra prevented liver failure and reduced mortality in rats with FHF. These animals also exhibited improved liver function and increased survival rates after injection with these cells. Using green fluorescent protein as a marker, we demonstrated that the engrafted cells and their progeny were incorporated into injured livers and produced albumin. This study suggests that AF-MSCs genetically modified to over-express IL-1Ra can be implanted into the injured liver to provide a novel therapeutic approach to the treatment of FHF.

Rare inborn errors associated with chronic hepatitis B virus infection

Chronic hepatitis B (CHB) is a major global health issue. The role of rare genetic variants in CHB has not been elucidated. We aimed to identify rare allelic variants predisposing to CHB. We performed exome sequencing in 50 CHB patients who had no identifiable risk factors for CHB and 40 controls who were healthy and hepatitis B surface antibody‐positive, but had never received hepatitis B vaccination. We selected six rare variant alleles and followed up their association with disease status by Sanger sequencing in a case‐control study comprising 1,728 CHB patients and 1,636 healthy controls. The latter had either not been immunized with hepatitis B vaccine or had uncertain vaccination status. Our results showed that transmembrane protein 2 p.Ser1254Asn, interferon alpha 2 p.Ala120Thr, its regulator NLR family member X1 p.Arg707Cys, and complement component 2 p.Glu318Asp were associated with CHB, with P values of <1.0 × 10−7, 2.76 × 10−5, 5.08 × 10−5, 2.78 × 10−4 and odds ratios (ORs) of 2.45, 4.08, 2.34, and 1.97, respectively. The combined P value was <2.0 × 10−16. As there has been no indication of immunological functions for the associated gene, transmembrane protein 2, we further studied its expression by immunohistochemistry, real‐time polymerase chain reaction, and western blotting. Our results showed that it was strongly expressed by healthy hepatocytes, but its expression was reduced in liver tissues with CHB, hepatitis B viral (HBV) genome‐containing HepG2.2.15 cells, as compared with healthy liver tissues and non‐HBV genome‐containing HepG2 cells (P = 0.022 and 0.0036, respectively). Conclusion: We identified four missense mutations associated with CHB, our results providing evidence for rare inborn genetic defects that contribute to increased host susceptibility to CHB. (HEPATOLOGY 2012;56:1661–1670)

High level of IL-27 positively correlated with Th17 cells may indicate liver injury in patients infected with HBV

Interleukin‐6/IL‐12 family cytokines play a key role in inflammatory diseases via their effects on the differentiation or regulation of T helper cells.

Analysis of Monocytic and Granulocytic Myeloid-Derived Suppressor Cells Subsets in Patients with Hepatitis C Virus Infection and Their Clinical Significance

Myeloid-derived suppressor cells (MDSCs) have been shown to inhibit T-cell responses in many diseases, but, in hepatitis C virus (HCV) infected patients, MDSCs are still poorly studied. In this assay, we investigated the phenotype and frequency of two new populations of MDSCs denoted as monocytic and granulocytic MDSCs (M-MDSCs and G-MDSCs) in HCV infected patients and analyzed their clinical significance in these patients respectively. We found that the frequency of CD14+HLA-DR−/low cells (M-MDSCs) from HCV infected patients (mean ± SE, 3.134% ± 0.340%) was significantly increased when compared to healthy controls (mean ± SE, 1.764% ± 0.461%) (Z = −2.438, P = 0.015), while there was no statistical difference between the frequency of HLA-DR−/lowCD33+CD11b+CD15+ (G-MDSCs) of HCV infected patients and healthy donors (0.201% ± 0.038% versus 0.096% ± 0.026%, P > 0.05), which suggested that HCV infection could cause the proliferation of M-MDSCs instead of G-MDSCs. Besides, we found that the frequency of M-MDSCs in HCV infected patients had certain relevance with age (r = 0.358, P = 0.003); patients older than 40 years old group (mean ± SE, 3.673% ± 0.456%) had a significantly higher frequency of M-MDSCs than that of age less than 40 years old group (mean ± SE, 2.363% ± 0.482%) (Z = −2.685, P = 0.007). The frequency of M-MDSCs, however, had no correlation with HCV RNA loads, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and the level of liver inflammation degree.

Peginterferon-α2a combined with response-guided short-term lamivudine improves response rate in hepatitis B e antigen-positive hepatitis B patients: a pilot study.

Aims The hepatitis B e antigen (HBeAg) seroconversion rate in HBeAg-positive chronic hepatitis B patients treated with peginterferon-&agr;2a (peg-IFN &agr;2a) is still low (about 30%). The aim of this study was to find a new combination therapy of peg-IFN &agr;2a with lamivudine to improve the efficacy in HBeAg-positive chronic hepatitis B patients. Patients and methods All patients started with peg-IFN &agr;2a treatment at a dose of 135 &mgr;g/week. If the concentration of hepatitis B virus (HBV) DNA was greater than 1.0×104 copies/ml and if the patient was positive for HBeAg at 12 weeks of treatment, lamivudine was included into the treatment for 12 weeks. Thereafter, the patients continued on peg-IFN &agr;2a alone for the full 52-week treatment course. Results Thirty-two patients were recruited, and eight of them achieved HBV DNA concentrations of less than 1.0×104 copies/ml or HBeAg loss at 12 weeks of treatment when lamivudine was not administered (group A). The other 24 patients received additional lamivudine, started from 12 weeks of treatment for 12 weeks (group B). At the end of treatment (EOT), in the peg-IFN &agr;2a monotherapy group (group A), eight patients (100%) had HBV DNA loss, six patients (75%) achieved HBeAg seroconversion, and eight patients (100%) achieved alanine aminotransferase (ALT) normalization. This level of response was sustained for 24 weeks after treatment in all patients with an early response. In the peg-IFN &agr;2a combined short-term lamivudine group (group B), 12 patients (50%) had HBV DNA loss, nine patients (38%) achieved HBeAg seroconversion, one patient (4%) achieved hepatitis B surface antigen loss, and 15 patients (63%) achieved ALT normalization at EOT. One patient had an HBV DNA rebound and an HBeAg reversion 24 weeks after treatment. The total HBV DNA loss rate, HBeAg seroconversion rate, hepatitis B surface antigen loss rate, and ALT normalization rate in all patients were 59, 47, 3, and 69%, respectively, at EOT and were 56, 44, 3, and 69% 24 weeks after treatment, respectively. Conclusion This study indicates that the response-guided approach resulted in an overall HBeAg seroconversion rate of 47% at EOT and 44% 24 weeks after treatment. This promising strategy to increase response rates with peg-IFN &agr;2a warrants further investigation.

Tumoral indoleamine 2, 3-dioxygenase 1 is regulated by monocytes and T lymphocytes collaboration in hepatocellular carcinoma

Indoleamine 2, 3-Dioxygenase 1 (IDO1) in cancer cells plays a critical role in tumor immunosuppression. However, the precise mechanisms regulating tumoral IDO1 expression in tumor milieus remain unclear. Here, we reported that IDO1 expression in tumor cells of hepatocelluar carcinomas (HCC), displayed a discrete rather than uniform pattern. In vitro culture, human hepatoma cell lines did not constitutively express IDO1. Interestingly, co-culture with peripheral blood mononuclear cells (PBMC) significantly induced and maintained IDO1 expression in these tumor cells, predominantly through IFN-γ. Mechanistically, we showed that IDO1 expression in tumor cells was only induced when co-cultured with both T lymphocytes and monocytes. Moreover, the cooperation between T lymphocytes and monocytes played an indispensable role on the tumoral IDO1 expression in immunocompromised mice. Taken together, our data supported the notion that IDO1 expression in tumor cells might serve as a counter-regulatory mechanism regulated by immune system, and provided new insights into the collaborative action of different inflammatory cells in tumor immunosuppression.

Prevalence of IL-28B and ITPA genotypes in Chinese Han population infected persistently with hepatitis C virus genotype 6 or HCV-1

The geographic distribution, demographics, epidemiology, host factors, and clinical characteristics of persistent HCV‐6 infection in China need further characterization. This multicenter study enrolled 63 patients with persistent HCV‐6 infection and 63 patients with persistent HCV‐1 infection as controls. Blood biochemistry, quantitation of HCV RNA levels, and identification of host IL‐28B genotypes (rs12979860, rs8099917, and rs12980275) and ITPA genotype (rs1127354) were performed to estimate potential variability in host factors that may affect response to treatment. The mean HCV‐6 RNA level (3.8E6 IU/ml) was significantly higher than that in patients infected with HCV‐1 (1.7E6 IU/ml; P < 0.001). Patients persistently infected with HCV‐6 had a high prevalence of IL‐28B rs12979860 CC genotype (92.1%), rs8099917 TT genotype (93.7%), and rs12980275 AA genotype (90.5%). Their prevalence in patients infected with HCV‐1 was only modestly lower (82.5%, 84.1%, and 82.5%, respectively; P > 0.05). The inosine triphosphate pyrophosphatase (ITPA) SNP rs1127354 CC genotype was present in 66.7% of patients infected with HCV‐6, comparable to that of patients infected with HCV‐1 (65.1%; P > 0.05). There were no differences in the liver function, proportion of hepatic cirrhosis patients or patients with increased serum glucose between these two groups. Persistent HCV‐6 infection in Chinese Han is found mainly in the southern China. Chinese Han with chronic HCV‐1 or HCV‐6 infection have IL‐28B genotypes, suggesting responsiveness to interferon‐based pharmacotherapy. Most patients (67%) possess the ITPA genotype associated with susceptibility to ribavirin‐induced hemolysis. The routes of transmission for HCV‐6 genotype were more diversified than HCV‐1 genotype. The outbreak of HCV‐6 infection through blood transfusion progressed faster than HCV‐1. J. Med. Virol. 85:1163–1169, 2013.

Dynamic Changes of the Frequency of Classic and Inflammatory Monocytes Subsets and Natural Killer Cells in Chronic Hepatitis C Patients Treated by Direct-Acting Antiviral Agents

Objective Up to now, little was known about the immunological changes of chronic hepatitis C (CHC) patients treated with direct-acting antiviral agents (DAAs); we try to explore the effect of DAAs on the frequency of monocytes, NK cells, and cytokines that promote their activation. Methods 15 treatment-naive CHC patients and 10 healthy controls were recruited. Patients were examined before DAAs therapy (0 w) and at week 4 (4 w) and week 12 (12 w) of therapy. Percentage of monocytes and NK cells of the peripheral blood was analyzed by flow cytometry. Serum cytokines IL-12, IL-18, CXCL10, CXCL11, sCD14, and sCD163 were measured by enzyme linked immunosorbent assay. Results The frequency of CD3–CD16+CD56+ NK cells and classic CD14++CD16− monocytes decreased, while CD14+CD16+ monocytes and cytokines IL-12, IL-18, CXCL10, CXCL11, sCD14, and sCD163 increased at 0 w compared to healthy controls. During DAAs treatment, the decreased NK cells and classic monocytes gradually increased to normal levels; the increased inflammatory monocytes and cytokines IL-12 and CXCL11 decreased to normal levels, but the increased cytokines IL-18, CXCL10, sCD14, and sCD163 still remained at high levels at 12 w though they decreased rapidly from 0 w. Conclusion Our results showed that DAAs treatment attenuated the activation of monocytes and NK cells in CHC patients. Trial registration number is NCT03063723.

Pretreatment HBsAg level and an early decrease in MELD score predict prognosis to lamivudine treatment for HBeAg-negative acute-on-chronic liver failure.

BACKGROUND AND OBJECTIVE Few data are available about the predictability of HBsAg quantification to nucleos(t)ide analogues treatment in acute-on-chronic liver failure (ACLF). The aim of this study was to investigate HBsAg level combined with the model for end-stage liver disease (MELD) score for predicting prognosis to lamivudine monotherapy in HBeAg-negative ACLF. METHODS Fifty-seven nucleoside-naïve patients with HBeAg-negative ACLF were treated with 100mg of lamivudine daily. Serum levels of HBsAg, HBV DNA and biochemical items were detected at baseline, before death (patients died within 3months) or month 3 meanwhile MELD score was calculated. Dynamic of these items and 3-month mortality were analyzed. RESULTS HBV DNA level significantly decreased while HBsAg level did not after treatment. Twenty-six patients died within 3months and the others survived. Regardless pre- or post-treatment, HBsAg level of survival group was significantly higher than that of dead group meanwhile MELD scores of the former were significantly lower than those of the latter (all P<0.05). Post-treatment MELD scores of 32 patients with pretreatment HBsAg levels above 4000 COI were significantly lower than those of 25 patients below to it (t=-2.116, P=0.044) and the 3-month mortality of the formers was significantly lower than that of the latter (34.3% [11/32] vs 64.0% [16/25], χ(2)=4.941, P=0.026). CONCLUSIONS In HBeAg-negative ACLF, patient with higher pretreatment HBsAg levels and early decrease in MELD score has lower 3-month mortality than one without it during lamivudine monotherapy.

A Novel prognostic scoring system to predict 3-month mortality risk in patients with acute-on-chronic liver failure in hepatitis B: a retrospective cohort study

PurposeThe present study was done to establish an objective, sensitive prognostic scoring system and to determine the applicability of this model in predicting the 3-month mortality of patients with acute-on-chronic liver failure in hepatitis B (ACLFB).MethodsWe developed a novel prognostic scoring system, calculated from six clinical indices including serum total bilirubin, prothrombin activity, serum creatinine, hepatic encephalopathy, infections, and the depth of ascites from 499 patients with ACLFB. Differences in the sensitivity, specificity, and practicality of a Novel prognostic scoring system and the model of end-stage liver disease (MELD) were analyzed.ResultsThe areas under the receiver operating characteristic curve (ROC) for the Novel scoring systems and MELD scoring systems were 0.967 (95% CI, 0.956–0.977) and 0.900 (95% CI, 0.878–0.922), respectively. The analysis of the ROC curve indicated that the Novel scoring systems were an exact, pertinent, and objective prognostic model with greater accuracy than the MELD. In the Novel scoring systems, the survival rate of these patients whose scores ranged from 2 to 6 was 98.80%, while for those whose scores point at 7 and 15, the mortality rates were 8.70% (2/23) and 95.45% (21/22), respectively, and the mortality rate of these patients whose scores were 16 and above was 100.00%. However, in the MELD prognostic scoring systems, there were no score ranges with 100.00% survival rate.ConclusionsWe developed an objective, pertinent, and sensitive prognostic scoring system that predicted the 3-month mortality of patients with ACLFB with greater accuracy than the MELD.