Zhan-Lian Huang

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.

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.

Efficacy of sequential use of telbivudine in hepatitis B e antigen-positive chronic hepatitis B patients with partial responses to pegylated interferon: a pilot study

The aim of this study was to investigate the efficacy of sequential use of telbivudine in hepatitis B e antigen (HBeAg)‐positive chronic hepatitis B patients with partial responses to pegylated interferon. Patients with partial responses to 48 weeks of pegylated interferon treatment were divided into two groups. In group A, patients stopped pegylated interferon directly without sequential treatment. In group B, patients received sequential treatment with telbivudine 600 mg/day. HBeAg, HBeAb, hepatitis B virus (HBV) DNA, alanine aminotransferase (ALT) and creatine kinase levels were determined at baseline and at weeks 12, 24, 36 and 48. Responses and safety were assessed after 48 weeks of telbivudine treatment. Thirty‐six patients were recruited. Eighteen of these patients stopped pegylated interferon without sequential treatment (group A). After 48 weeks of follow‐up, five patients (28%) had undergone HBeAg seroconversion, nine patients (50%) had undetectable levels of HBV DNA, and 11 patients (61%) achieved normal alanine aminotransferase (ALT) levels. The other 18 patients received sequential telbivudine treatment (group B). After 48 weeks of treatment, 11 patients (61%) had undergone HBeAg seroconversion, and all patients had undetectable levels of HBV DNA and normal ALT levels. All patients tolerated sequential telbivudine treatment, and only slightly elevated creatine kinase levels were observed. Switching to telbivudine therapy was efficient and safe in HBeAg‐positive chronic hepatitis B patients with partial responses to 48 weeks of pegylated interferon. Sequential treatment with telbivudine resulted in an HBeAg seroconversion rate of 61% and an HBV DNA loss rate of 100% after 48 weeks. This promising strategy warrants further investigation.

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.

Liver myofibroblasts up-regulate monocyte CD163 expression via PGE2 during hepatitis B induced liver failure

BackgroundAlthough patients with liver failure exhibit a generalized inflammatory-imbalance status, substantial evidence indicates that this immunosuppressive or anti-inflammatory state may be deleterious. Increased expression of CD163 (known to be involved in several anti-inflammatory functions of the immune system) in patients with liver failure is significantly correlated with a fatal outcome. However, little is known of the regulatory mechanisms that influence the expression of CD163.MethodsWe assessed the expression of CD163 on monocytes from both circulating cells and the liver tissues of patients with hepatitis B induced liver failure using flow cytometry and isolated the myofibroblasts from diseased livers. The ability of human liver myofibroblasts to regulate CD163 expression on monocytes was studied in vitro.ResultsWe showed that CD163+ monocytes were enriched primarily in diseased livers and that they were associated with liver myofibroblasts in the same area. Accordingly, liver myofibroblasts were significantly superior to normal skin fibroblasts in inducing the expression of CD163 on monocytes in vitro. Moreover, we found that liver myofibroblasts triggered the activation of monocytes by secreting PGE2. Inhibition of PGE2 production in liver myofibroblasts using NS-398 markedly reduced CD163 expression in vitro.ConclusionThese results suggest that liver myofibroblasts play a direct role in regulating the expression of CD163 on monocytes in human liver tissues and thereby may regulate monocyte function during hepatitis B induced liver failure.

BTLA identifies dysfunctional PD-1-expressing CD4(+) T cells in human hepatocellular carcinoma.

ABSTRACT Although immunotherapy targeting programmed cell death 1/programmed cell death ligand 1 (PD-1/PD-L1) pathway is being applied in clinic, the response outcomes are heterogeneous, suggesting existences of distinctive subsets within PD-1-expressing T cells that react differently to PD-1/PD-L1 blockade. However, markers to demarcate these subsets in human cancers remain unclear. Here, we found that both PD-1 and B and T lymphocyte attenuator (BTLA) were significantly upregulated on CD4+ T cells from tumor compared with those from paired non-tumor liver in hepatocellular carcinoma (HCC) patients. Interestingly, over 85% BTLA+ CD4+ T cells were PD-1-expressing cells and represented about 50% PD-1+ CD4+ T cells in tumors, and that level of BTLA+PD-1+ tumor CD4+ T cells were selectively associated with advanced stage HCC. BTLA+ identified highly dysfunctional PD-1-expressing CD4+ T cell subset, whereas BTLA− defined PD-1+ CD4+ T cells undergoing activation in HCC. Importantly, blockade of PD-L1 could restore the ability of IFNγ/TNF-α production in BTLA+PD-1+ tumor CD4+ T cells but partially suppressed the activation of BTLA−PD-1+ CD4+ T cells. Moreover, we provided evidence that BTLA signals also participated in suppressing CD4+ T cell function in HCC. In conclusion, BTLA could identify distinct function of PD-1 expressing CD4+ T cells in human cancer, which might not only advance our understanding of inhibitory receptor blockade, but also provide new targets for clinical predictors of response to these immunotherapies.

Intrahepatic NK cells function suppressed in advanced liver fibrosis

Although numerous epidemiological studies indicate that hepatitis B virus‐related liver fibrosis (HBV‐LF), particularly cirrhosis, represents the main risk factor for liver cancer development, the mechanisms determining the persistence of fibrosis and liver cancer pathogenesis are still poorly defined. Few studies have investigated the status of NK cells during different stages of HBV‐LF.

An albumin, collagen IV, and longitudinal diameter of spleen scoring system superior to APRI for assessing liver fibrosis in chronic hepatitis B patients

OBJECTIVES The aim of this study was to screen the non-invasive indexes correlated with liver fibrosis and establish a scoring system for the diagnosis of liver fibrosis in hepatitis B patients. METHODS Data of 34 non-invasive indexes were collected for 208 hepatitis B patients. Correlation analysis and stepwise discriminant analysis was used to screen out indexes useful for the diagnosis of liver fibrosis. Finally, a scoring system composed of indexes screened out by stepwise discriminant analysis was established for the assessment of liver fibrosis. RESULTS Twenty-one indexes correlating with liver fibrosis were screened out by correlation analysis; hyaluronic acid had the highest r-value, 0.456. A scoring system including albumin, collagen IV, and the longitudinal diameter of the spleen was established. The areas under the receiver operating characteristic curves (AUC) for this scoring system and the aspartate aminotransferase to platelet ratio index (APRI) in differentiating S3-4 from S0-2 were 0.79 (95% confidence interval (CI) 0.72-0.85) and 0.27 (95% CI 0.18-0.35), respectively. With a cut-off value of <3, the presence of significant fibrosis (S3-4) could be excluded by this scoring system with a negative predictive value of 86.1% and sensitivity of 86.8%. With a cut-off of >6, the presence of S3-4 fibrosis could be correctly identified with a positive predictive value of 73.6% and specificity of 87.6%. Using this scoring system, 53.4% of patients could be classified correctly and avoid liver biopsy. CONCLUSIONS The scoring system provides a simpler method to identify significant fibrosis (S3-4) in chronic hepatitis B patients.

The paradoxical changes of membrane and soluble herpes virus entry mediator in hepatocellular carcinoma patients

The herpes virus entry mediator (HVEM) network has become new directions in targeting novel checkpoint inhibitors for cancer therapy. However, the changes of membrane‐bound HVEM (mHVEM) and soluble HVEM (sHVEM) in hepatocellular carcinoma (HCC) are not fully understood. This study aims to study the changes of mHVEM and sHVEM in HCC patients.