| Background and Objectives:According to the world health organization’s report, there were about2billion people had been affected by hepatitis B virus (hepatitis B virus, HBV) all over the world. Among them,350million people were with chronic HBV infection and each year about1million people died from chronic liver failure, posthepatitic cirrhosis and primary hepatocellular carcinoma which was related with chronic HBV infection. Prevalent intensity of HBV infection has a great difference among different areas of the world, our country has been reduced to the medium endemic area. The national epidemiological survey of2006showed that positive rate of hepatitis B surface antigen in the general population of our residents from1to59years was about7.18%, while that of children under the age of five was only1%. Therefore, it is predicted that there are about93million existing chronic HBV infection cases in our country, about20million of whom are chronic hepatitis B patients. Chronic hepatitis B infection has been a serious threat to human health.Most of the HBV infection presents a subclinical infection process; a small part performs acute viral hepatitis, the virus will soon be cleared; very small part develops into fulminant liver hepatitis; another small part changes into chronic hepatitis, recurs repeatedly and last for many years, some of whom eventually turns into hepatitis cirrhosis or primary hepatocellular carcinoma. The mechanism of causing chronic HBV infection to different outcomes is not clear. Researches show that among HBV genome coded proteins, only hepatitis B x protein(HBx) has a variety of regulation and control function, which determines the complex function of HBV X gene and its expressive product HBx in immunologic mechanism of HBV infection. HBx has complex biological activities, such as trans-active function to a variety of genes and closely related to integration of virus gene and mutation of host cell gene. A wide range of influences might be accomplished through a variety of regulation and control of X protein over replication and proliferation of the virus as well as process of apoptosis and canceration of host cell directly or indirectly. HBx encoded by HBV X gene contains six areas (A～F), with a total of154amino acids (amino acid, aa). HBx protein encoded by HBV genome exerts a variety of regulations and controls, which include activating the transcription of host cells and viral gene itself, regulating apoptosis, inhibiting circumscribed repairing reaction of damaged DNA of cell, enhancing transcription activity of the original cancer gene, activating telomerase reverse transcriptase and pathways of cell signal transduction, promoting liver fibrosis etc. HBx Antigen(HBxAg) presents a high expressive state in hepatocellular tissues, which may trans-activate abnormal activation of liver cell proto-oncogene, and promote cell apoptosis and canceration process. HBV X gene and HBx protein have become a hot research topic at home and abroad recently.Although most hepatitis B viruses live in liver cells, some may infringe upon the host immune system, activate the immune cells, and induce inflammation and antibody production. The antibody induced by HBxAg is called HBxAb(also known as anti-HBx). Park EH et. al. reported that the HBxAg was detected in liver tissues of chronic HBV infection, liver cirrhosis and hepatocellular carcinoma(HCC). Horiike N etc. found HBxAg existed in liver tissue of patients with chronic viral hepatitis B generally and had significant correlation with alanine transaminase while serum markers of HBV replication had no relevance.Huang Y, etc. found that the quantity of HBxAg and HBeAg were similar in serum of patients with chronic hepatitis; In patients with the serum HBeAg, HBVDNA and liver tissue HBcAg positive, HBxAg has a high positive rate.The detection of expression of HBV X gene might be helpful to the early diagnosis and prognosis judgment of chronic hepatitis and the generated liver cirrhosis as well as cancer.In regard to the antibody (anti-HBx) against HBxAg, some scholars think that it is a general and early marker and appears shortly in patients with acute viral hepatitis B, but exists continually in chronic viral hepatitis B. Amount of anti-HBx relates with the duration and intensity of replication of hepatitis B virus and has no correlation with the severity of the liver illness and its malignant progress. Moriarty, etc. found anti-HBx was closely related with carcinoma related to HBV infection, and might be used as a predictor for liver cancer. Serum anti-HBx was also considered to be the mark of HBV replication, especially for cirrhosis and liver cancer related to hepatitis b virus, not a protective antibody.Some scholars applied targeting treatment in2patients with hepatocellular carcinoma with rat monoclonal anti-HBx as a carrier to carry iodine131, their AFP reduced significantly and the symptoms improved obviously after the therapy. Kim JW., etc. thought the existence of HBxAg/anti-HBx in process of chronic hepatitis b virus infection, were linked with serological mark of continuous duplication of hepatitis B virus.Many researches were published in the past decade on the relationship between hepatitis B virus X gene and hepatocellular carcinoma. Some were focused on studying host immune state and various cytokines in different developmental stages of chronic hepatitis B virus infection. Nevertheless, no reports was noticed on a direct comparison between anti-HBx and cytokines, and the relationship between anti-HBx and cytokines as well as the significance of the occurrence of anti-HBx in serum of cases with chronic hepatitis B virus infection was not clear. Whether the occurrence of anti-HBx affects the expression of immune factors of the host remains indistinct. In the present study, we detected anti-HBx, soluble Fas (Fas), IL-10and IL-12in serum of90patients in various developmental stages of chronic hepatitis B virus infection, including cases in immune-tolerant phase of HBV infection(i.e. asymptomatic carriers, ASC), immune clearance period (i.e. chronic hepatitis B, CHB), hepatitis B infection related liver cirrhosis(LC) and hepatocellular carcinoma (HCC), with the aim of exploring the mutual relations between anti-HBx and other immune index, clarify relations of changes in anti-HBx and prognosis of the disease, enrich hepatitis B pathogenic theory and provide basis of looking for the strategy, new targets and opportunity in anti-HB V treatment.Methods:Ninety cases with chronic HBV infections from Shandong province of China were randomly selected, including10of asymptomatic carriers,28of chronic hepatitis B,26of liver cirrhosis and26of hepatocellular carcinoma. Fasting blood was exsanguinated and serum was routinely prepared. Alanine transaminase (ALT), bilirubin (TBIL), and other liver function indexes were tested with automatic biochemical analyzer, double antibody sandwich enzyme-linked immunosorbent assays were applied for detection of IL-10, IL-12and soluble Fas(Fas), and the anti-HBx in serum was determined by the ELISA method established in our previous study.Results:1. IL-10, IL-12and Fas levels in peripheral blood trended to increase gradually from the asymptomatic carriers to chronic hepatitis B, liver cirrhosis and liver cancer patients; test results of the4groups of patients were13.93±14.40ng/L,39.38±20.77ng/L,69.06±46.37ng/L,62.82±23.42ng/L for IL-10;15.64±23.04ng/L,68.50±23.14ng/L,76.83±12.82ng/L,8383.74±24.88ng/L for IL-12; and58.17±77.42ng/L,179.88±104.36ng/L, 249.22±107.80ng/L, and252.98±87.65ng/L for Fas respectively. There was a significant difference over the above three items between asymptomatic carriers and other three groups (P<0.001); the group of liver cirrhosis and liver cancer were also higher than that of chronic hepatitis B respectively, IL-10and Fas level presented a significant difference (P<0.05), IL-12level between chronic hepatitis B and cirrhosis group had no statistically significant difference (P=0.154), and that between chronic hepatitis B group and liver cancer group had a significant difference (P=0.01). A comparison between liver cirrhosis group and liver cancer group showed no significant difference over the3items (P>0.05), however, both IL-12and Fas had a higher tendency in the latter, while IL-10had a little bit higher tendency in the former.2. Twenty-seven of90patients were anti-HBx positive, with1case of asymptomatic carriers (ASC),4cases of chronic hepatitis B (CHB),12cases of liver cirrhosis (LC) and10cases of hepatocellular carcinoma (HCC), and the positive ratio are10%,14.28%,46.15%and38.46%respectively.3. According to the qualitative result of anti-HBx, patients were grouped into anti-HBx negative and positive groups. The three cell factors, IL-10, IL-12and Fas levels of anti-HBx positive group(238.25±118.33ng/L,65.73±43.42ng/L and80.23±29.69ng/L respectively) were higher than that of the negative group(194.21±111.64ng/L,45.98±29.54ng/L and64.81±27.57ng/L respectively). IL-10and IL-12between2groups had a significant difference (P<0.05), while Fas results between them has no statistical significance (P=0.094).Conclusion:1. IL-10and IL-12levels in peripheral blood trend to increase gradually from the asymptomatic carriers to chronic hepatitis B, liver cirrhosis and liver cancer patients. However, an obviously increased secretion of IL-10over IL-12(i.e., IL-12being relative shortage) indicate that promotion of IL-10level is closely related to disease progression.2. Serum level of Fas maybe play a very important role in the process of the liver cirrhosis and liver cancer development, and measurement of serum Fas may be used as one of evaluation indexes for damage degree of liver tissue of patients with chronic hepatitis B infection.3. Serum anti-HBx is not protective antibody. IL-10, IL-12and Fas levels in anti-HBx positive patients are higher than that of the negative group and consistent with progressive degree of the disease. Anti-HBx in serum may be another serum indicator for the development from chronic hepatitis B to liver cirrhosis or liver cancer. |
PDF Full Text Request