| Objective:1. To analyze intrahepatic Treg cell levels in patients who were in the immune tolerant(IT) phase of chronic HBV infection and explore whether Treg cell is the key factor in leading to a continuous IT state of patients with chronic HBV infection. In order to provide theoretical basis for breaking the IT state of chronic HBV infection.2. To analyze the relationship between intrahepatic Treg and Th17 cell levels and the falling range of HBs Ag, HBe Ag and HVB DNA levels in peripheral blood of patients who were in the immune clearance(IC) phase of chronic HBV infection at two weeks. And to explore how the intrahepatic Treg cell levels influnce the organism ability to clear HBV by autoimmune response. In order to provide theoretical reference for improving the capacity of organism to clear HBV.3. To analyze the change of intrahepatic Treg/Th17 balance in patients who were in IC phase of chronic HBV infection and with different degree of inflammation and fibrosis. And to explore how the change of Treg/Th17 balance influnce the progression of disease, in order to provide theoretical basis for reducing the risk of developing liver failure or liver cirrhosis.Methods:68 patients with chronic HBV infection and underwent liver biopsy were included. The 68 patients included 20 in IT phase, 36 in IC phase and 12 in inactive phase. Immune histochemical method was employed to count the intrahepatic Treg/Th17 cell levels. The HBs Ag, HBe Ag and HVB DNA levels in peripheral blood were detected by ECLIA and real-time fluorescent quantitative PCR technique respectively. 8 healthy liver transplant donors were collected as controls. To analyze the change of Treg/Th17 balance in patients with different phase of chronic HBV infection and healthy controls. To analyze the relationship between intrahepatic Treg cell levels and the falling range of HBs Ag, HBe Ag and HVB DNA levels in peripheral blood in patients who were in the IC phase of chronic HBV infection at two weeks. To observe the change of Treg/Th17 balance in patients who were in IC phase of chronic HBV infection and with different degree of inflammation andfibrosis and analyze the relationship between the Treg/Th17 balance and the degree of liver injury.Results:1. The intrahepatic Treg cell levels increased without significant difference in IT phase and inactive phase group compared with healthy controls [respect ively,(2.44±0.65)cells/hpf,(3.37±1.02)cells/hpf,(1.13±0.21)cells/hpf; all P>0.05].The intrahepatic Treg cell levels[(13.90±5.12)cells/hpf] increased significantly in IC phase group compared with the other three groups(all P<0.01). There wa s no significant difference between IT phase group and inactive phase group(P>0.05). The Treg cell levels in IC phase group with different degree of inflammation and fibrosis had significant differences: G4 group > G3 group >G2 group [respectively,(22.08±0.66)cells/hpf,(16.90±1.47)cells/hpf,(9.85±2.79)cells/hpf; all P<0.001]; S3 group > S2 group > S1 group [respectively,(21.77±0.97)cells/hpf,(14.64±2.66)cells/hpf,(7.70±1.20)cells/hpf; all P<0.001].2. The HBs Ag, HBe Ag and HVB DNA levels in peripheral blood at two weeks correlated negatively with the intrahepatic Treg cell levels in patients in IC phase of chronic HBV infection(P<0.001), but there was no correlation between Th17 cell and items mentioned above(P>0.05).3. Compared with healthy controls, the intrahepatic Th17 cell levels increased without significant difference in IT phase group(P>0.05), but in IC phase and inactive phase group the intrahepatic Th17 cell levels increased significantly[respectively,(1.08±0.18)cells/hpf,(1.98±0.55)cells/hpf,(17.37±1.52)cells/hpf,(3.82±0.67)cells/hpf; all P<0.05]. The intrahepatic Th17 cell levels increased significantly in IC phase group and inactive phase group compared with IT phase group(all P<0.001). The intrahepatic Th17 cell levels increased significantly in IC phase group compared with inactive phase group(all P<0.001). The Th17 cell levels in IC phase group with different degree of inflammation and fibrosis had significant differences: G4 group > G3 group > G2 group [respectively,(20.12±0.54)cells/hpf,(17.50±1.13)cells/hpf,(16.56±0.92)cells/hpf; all P<0.05]; S3 group > S2 group > S1 group [respectively,(20.03±0.53)cells/hpf,(17.21±0.80)cells/hpf,(16.08±0.93)cells/hpf; all P<0.05].4. Compared with healthy controls, the intrahepatic Treg/Th17 ratio increased without significant difference in IT phase group(P>0.05), but decreased in IC phase group significantly(P<0.05), and in inactive phase group, the intrahepatic Treg/Th17 ratio decreased without significant difference(P>0.05). The intrahepatic Treg/Th17 ratio in IC phase group and inactive phase group decreased significantly compared with IT phase group(all P<0.01), and there was no statistically significant difference between IC phase group and inactive phase group(P>0.05). The intrahepatic Treg/Th17 ratio in IC phase group with different degree of inflammation and fibrosis had significant difference: G4 group > G3 group > G2 group [respectively,(1.10±0.02),(0.97±0.39),(0.59±0.15); all P<0.05]; S3 group > S2 group > S1group[respectively,(1.09±0.03),( 0.85±0.15),(0.48±0.05); all P<0.001].Conclusion:1. No significant changes were observed in Treg and Th17 cell levels and their balance in patients who were in IT phase of chronic HBV infection compared with healthy controls, which suggested that the continuous IT state of patients with chronic HBV infection didn’t result from aggregation of a large number of Treg cells in liver or Treg/Th17 imbalance. So the IT mechanism remains to be further research.2. The intrahepatic Treg cell levels increased significantly in patients who were in IC phase of chronic HBV infection. And more Treg cells were found to be infiltrated in the livers of patients with a higher G/S score than those of patients with a lower G/S score, which suggested that abundant Treg cells in liver may be able to inhibit the organismic immune response to protect the host from serious pathological damage caused by a strong immune response in patients who were in IC phase of chronic HBV infection. In addition, the HBs Ag, HBe Ag and HVB DNA levels in peripheral blood at two weeks negatively correlated with the intrahepatic Treg cell levels in patients who were in the IC phase of chronic HBV infection, which suggested that abundant Treg cells may limit the organismic capacity to clear HBV by autoimmune response, and then influence the eradication of HBV.3. The intrahepatic Th17 cell levels increased significantly in patients who werein IC phase of chronic HBV infection. And more Th17 cells were found to be infiltrated in the livers of patients with a higher G/S score than those of patients with a lower G/S score, which suggested that Th17 cells may actively participate in the immune-mediated liver injury and the development of hepatic fibrosis in CHB patients.4. The intrahepatic Treg/Th17 ratio in patients who were in IC phase of chronic HBV infection and with different degree of inflammation and fibrosis had significant differences, and Treg/Th17 ratio increased with inflammation and fibrosis progression,which suggested that when the liver was slightly injured, Th17 cells increased more significantly than Treg cells, however when the liver was injured seriously, Treg cells increased more significantly than Th17 cells. So, when the liver was injured seriously,we may be able to reduce the risk of developing liver failure or liver cirrhosis by changing the Treg/Th17 balance with immunotherapy. |
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