Immune Netw.  2009 Feb;9(1):20-26. 10.4110/in.2009.9.1.20.

Increase of Plasma IL-12/p40 Ratio Induced by the Combined Therapy of DNA Vaccine and Lamivudine Correlates with Sustained Viremia Control in CHB Carriers

• Im SJ
• Yang SH
• Yoon SK
• Sung YC

• KMID: 2167993
• DOI: http://doi.org/10.4110/in.2009.9.1.20

Abstract

BACKGROUND: We previously reported that IFN-gamma producing T cell responses induced by the combined therapy of DNA vaccine and lamivudine for one year are important for the induction of sustained virological response (SVR). However, IFN-gamma production is not sufficient to predict sustained viremia control in chronic hepatitis B (CHB) carriers treated.
METHODS
Twelve CHB carriers were intramuscularly immunized 12 times at a 4-week interval with 8 mg of HBV DNA vaccine during the standard lamivudine treatment (100 mg/daily/1 year). The level of cytokines during and after the combined therapy in plasma of all 12 CHB carriers treated was determined by each ELISA kit. Six out of 12 CHB carriers revisited the clinic, and their HBV DNA levels were examined.
RESULTS
The combined therapy increased plasma IL-12 and IL-12/p40 ratio during the treatment (baseline vs. peak level: 41.8+/-8.3 vs. 163.1+/-29.2 pg/ml; p<0.01 and 0.96+/-0.25 vs. 3.58+/-0.86; p<0.01, espectively), and the peak level of plasma IL-12 and IL-12/p40 ratio was evoked at 6 to 10 months during the combined therapy. In particular, CHB carriers with SVR had two and three-fold higher level of the peak plasma IL-12 and plasma IL-12/p40 ratio than non-virological responders (NVRs), respectively (218.0+/-41.4 vs. 108.1+/-28.6 pg/ml; p=0.09 and 5.35+/-1.38 vs. 1.80+/-0.29; p<0.05, respectively), while p40 level was consistent during the combined therapy. In addition, there was no significant temporal correlation between the peak IL-12/p40 ratio and the elevation of serum alanine aminotransferase (ALT) in this study, contrast to IFN-alpha therapy which induced peak IL-12 level following ALT flares.
CONCLUSION
Our results indicate that the combined therapy induces the increase of plasma IL-12 and IL-12/p40 ratio, which are associated with long-term SVR in CHB carriers.

Keyword

hepatitis B virus; DNA vaccine; interleukin-12; IL-12/p40 ratio; sustained viremia control

MeSH Terms

Alanine Transaminase
Corynebacterium
Cytokines
DNA
Enzyme-Linked Immunosorbent Assay
Hepatitis B virus
Hepatitis B, Chronic
Interleukin-12
Lamivudine
Plasma
Viremia
Alanine Transaminase
Corynebacterium
Cytokines
DNA
Interleukin-12
Lamivudine

Figure

• 

  Figure 1 Experimental schedule of the pilot-clinical study. Lamivudine treatment for 8 weeks had been preceded before the first HB-100 injection at T1 except for three subjects; V#118 at week 4, and V#105 and V#106 at week 12. The vaccinees were intramuscularly administrated 12 times with 8 mg of HB-100 every 4 weeks as indicated by the arrows in combination with 100 mg of LAM treatment daily for 52 weeks (gray rectangles) and another 52 weeks were followed (white rectangles). Plasma samples were obtained every 4 weeks (black circles) and the level of cytokines was evaluated by ELISA assay.

• 

  Figure 2 (A, B) Serial analysis of the average level of plasma IL-12, p40 (A), and IL-12/p40 ratio (B) (mean±SEM) in CHB carriers during and after the combined therapy of DNA vaccine plus lamivudine. The level of plasma IL-12 and p40 was determined by a specific ELISA. (*p<0.05, **p<0.01 by Wilcoxon signed rank test) (C-E) The level of plasma IL-12 (C), p40 (D), and IL-12/p40 ratio (E) in each subject was shown before the treatment (pre, T0), the peak level detected during the combined therapy (peak, usually T6 to T10), and at the end of treatment (end, T12).

• 

  Figure 3 The level of peak plasma IL-12, p40 (A), and peak IL-12/p40 ratio (B) between virological responders (VRs) and nonvirological responders (NVRs) after combined therapy. Peak plasma IL-12 and peak IL-12/p40 were mostly detected from 6 (T6) to 10 (T10) months in 12 CHB carriers treated with combined therapy. The statistical analysis was done by Mann-Whitney U test.

• 

  Figure 4 The longitudinal analysis of the level of serum ATL and IL-12/p40 ratio in virological responders during the combined therapy. The kinetics of serum ALT levels (closed circles) and IL-12/p40 ratio (open circles) in VRs was compared in parallel to examine the temporal correlation between them.

• 

  Figure 5 The longitudinal analysis of serum viral loads for up to 3 years in CHB carriers treated with combined therapy. Three VRs (V#103, V#106 and V#113; closed circles) and 3 NVRs (V#105, V#107 and #114; open circles) out of 12 subjects revisited the clinic 3 years after the stopping the combined therapy. Data were represented by the average serum viral loads according to the time point. (mean±SEM).

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