Function of interleukin-17 and -35 in the blood of patients with hepatitis B-related liver cirrhosis

Abstract

Intrahepatic T helper (Th)17 cytokine and serum interleukin (IL)-17 levels in patients with hepatitis B are positively correlated with the progression of liver cirrhosis (LC). IL-35 can significantly inhibit the differentiation of Th17 cells and the synthesis of IL-17. The present study aimed to investigate the function and expression of IL-17 and IL-35 in the blood of patients with hepatitis B‑related LC. The levels of IL-17 and IL-35 in the peripheral blood of 30 patients with chronic hepatitis B (CHB), 79 with LC, 14 with chronic severe hepatitis B (CSHB), and 20 normal controls were detected by ELISA. Quantitative polymerase chain reaction was used to evaluate Epstein-Barr virus-induced gene 3 (EBI3), forkhead box (FOX)P3 and IL-17 mRNA expression levels in peripheral blood mononuclear cells (PBMCs). Western blotting was used to determine protein expression. The liver function of patients and normal controls was measured. EBI3, IL-17 and FOXP3 mRNA expression levels in PBMCs from patients with LC, CHB and CSHB were higher than those in cells from the controls. IL-17 mRNA levels differed significantly according to the Child-Pugh classification and exhibited an upward trend over time in contrast to a downward trend for EBI3 and FOXP3 mRNA. The changes in protein expression in the peripheral blood were consistent with the changes in mRNA expression. Serum IL-17 levels were positively correlated with total bilirubin (TBIL), alanine aminotransferase (ALT) and Child-Pugh grade, and were negatively correlated with albumin. These observed differences were significant. Serum IL-35 levels were negatively correlated with albumin, but not with Child-Pugh grade, ALT and TBIL. IL-17 and IL-35 may be critically involved in the pathogenesis of hepatitis B-related LC.

Figure 1

Fluorescent determination of EBI3, FOXP3 and IL-17 expression with polymerase chain reaction. NC, normal control; LC-A–C, liver cirrhosis Child-Pugh class A; LC-B, LC Child-Pugh class B; LC-C, LC Child-Pugh class C; CHB, chronic hepatitis B; CSHB, chronic severe hepatitis B, IL-17, interlaukin 17; EBI3, Epstein-Barr virus-induced gene 3; FOXP3, forkhead box P3.

Figure 2

Changes in expression of IL-35 (Epstein-Barr virus-induced gene 3), IL-17 and FOXP3 by western blotting. Lanes: 1, normal control; 2, LC Child-Pugh class A; 3, LC Child-Pugh class B; 4, LC Child-Pugh class C; 5, chronic hepatitis B; 6, chronic severe hepatitis B. IL, interleukin; FOXP3, forkhead box P3; LC, liver cirrhosis.

Figure 3

ELISA determination of IL-17 and IL-35 serum concentration. Serum concentrations of (A) IL-17 and (B) IL-35. IL, interleukin; NC, normal control; LC-A, liver cirrhosis Child-Pugh class A; LC-B, LC Child-Pugh class B; LC-C, LC Child-Pugh class C; CHB, chronic hepatitis B; CSHB, chronic severe hepatitis B.