[Induction of anti-hepatocellular carcinoma immunity by Hyper-IL-6 gene in vivo].

AIM: To prepare the mouse hepatocellular carcinoma (HCC) cells with the stable expression of Hyper-IL-6 and explore the possibility of inducing active anti-HCC immune response by Hyper-IL-6 gene. METHODS: Hyper-IL-6 gene was transfected into mouse HCC cells MM45T.Li using Lipofectamine(TM);2000. G418-resistant clones named MM45T-HIL-6 were selected and detected for the expression of Hyper-IL-6 gene by RT-PCR and ELISA. Mouse HCC cells transfected with pEGFP-C1, named MM45T-mock, were prepared as controls. Tumor models were established by injecting subcutaneously 5×10(5); cells of MM45T.Li, MM45T- mock and MM45T-HIL-6 on the right anterior limb of BALB/c mouse, respectively. In vivo experiments were performed to observe the tumorigenicity of MM45T.Li, MM45T-mock and MM45T-HIL-6. The levels of CD4(+); and CD8(+); T cells in mouse peripheral blood were detected by flow cytometry (FCM). RESULTS: RT-PCR and ELISA showed that Hyper-IL-6 gene was expressed in the MM45T-HIL-6 cells, but not in the control cells. We observed that the tumorigenicity of MM45T-HIL-6 decreased compared with control cells after they were inoculated subcutaneously into mice. FCM results indicated that the levels of CD4(+); and CD8(+); T cells in the peripheral blood significantly increased in the mice inoculated with MM45T-HIL-6 compared with the ones inoculated with MM45T.Li and MM45T-mock cells (P<0.05). CONCLUSION: The mouse HCC cells with the stable expression of Hyper-IL-6 can induce active anti-HCC immune response after inoculated subcutaneously into mice.

[Evaluation of effective doses of hepatitis B immunoglobulin to eliminate hepatitis B surface antigen from infected neonates].

OBJECTIVE: To determine the effective dose of hepatitis B immunoglobulin (HBIG) for clearing maternally-transmitted hepatitis B virus (HBV) from a newborn. METHODS: Full-term neonates born to HBV-infected mothers were tested for hepatitis B surface antigen (HBsAg) and HBV DNA in venous blood, Individuals with positive results within two hours after birth were selected for study, and divided among two treatment groups: research group receiving HBIG continually adjusted to quantitative levels of neonatal HBsAg and HBV DNA levels; control group receiving standard HBIG 200IU dose. All neonates were also treated with 10 micrograms of recombinant vaccine. The decreases in HBsAg and HBV DNA over 12 months were comparatively analyzed between the two treatment groups. RESULTS: The two treatment groups (HBIG adjusted vs. standard) were statistically similar in Apgar score (9.38+/-0.49 vs. 9.37+/-0.48), neonate body weight (3458.67+/-374.93 vs. 3558.61+/-322.85 g), maternal age (26.33+/-3.63 vs. 25.33+/-3.03), and initial HBsAg and HBV DNA levels (rank sum test Z = 1.381, and Z = 0.700, respectively) (all, P more than 0.05). Successful clearance of HBV infection within 12 months was achieved in significantly more neonates in the HBIG adjusted therapy group than in the standard therapy group (82.8% vs. 57.4%; x2 = 9.696, P less than 0.05). CONCLUSION: Adjusting the neonatal HBIG dose according to HBsAg and HBV DNA levels can improve the success rate of clearing maternally-transmitted HBV.

[The correlation of HBeAg expression and HBV-DNA in serum or peripheral blood mononuclear cells in patients with chronic hepatitis B].

OBJECTIVE: To study the relationship between HBeAg expression and HBV-DNA in serum and peripheral blood mononuclear cells (PBMCs). METHODS: 208 patients with chronic hepatitis B were included in this present study. HBV-DNA in the PBMCs were performed by polymerase chain reaction (PCR), with the serum HBV-DNA level being determined by the way of fluoresces quantities PCR (FQ-PCR). Meanwhile, HBV-GM was also detected via enzyme-linked immunosorbent assay (ELISA). RESULTS: There were 106 patients for positivity in the HBV-DNA level of PBMCs with 102 for negativity, in which the HBV-DNA high levels (HBV DNA load > or = 1.0E5) in serum were 91.5%, 45.1% (chi2=52.12, P>0.01) respectively, with 76.4% and 50.9% (chi2=21.55, P>0.01) for the positive percentage of HBeAg expression. CONCLUSION: A significantly positive correlation was found between HBV-DNA in PBMCs and serum HBV-DNA along with the positive percentage of HBeAg, indicating that obvious PBMCs' increase infected by HBV in patients with positivity of HBeAg and high level of serum HBV-DNA.

[Relationship between the HBV core gene mutation and the cellular immunity in host].

OBJECTIVES: To study the relationship between the mutation of Leu60Val in HBV core region and the cellular immunity in patients with chronic hepatitis B (CHB). METHODS: HBV DNA C gene mutation was confirmed by polymerase chain reaction (PCR) and sequencing the products directly. The cytokines (IFN-gamma, TNF-alpha and IL-2) levels in serum were measured by enzyme linked immunosorbent assay (ELISA). The distribution of T-lymphocyte subpopulations in peripheral blood was detected by flow cytometry (FCM). RESULTS: The mutation of Leu60Val was found in 19 out of the 91 CHB patients. With the CHB severity, the mutation rate was getting higher, especially in the severe hepatitis group. The IFN-gamma and TNF-alpha levels were much higher in mutant strain group than those in wild strain group (t=2.584, 4.766, P<0.01), so was the ratio of CD4+/CD8+ (t=2.275, P<0.05). CONCLUSION: The mutant strain of 60Val may increase affinity to HLA-I molecule, or up-regulate the expression of HLA-I molecule, resulting in the activation of CTL to release the cytokines and cause immune response in liver.