Correlation Between Circulating Natural Killer Cells And HLA-Cw Alleles In Hepatitis B Patients

ObjectiveTo examine the frequency of circulating natural killer cells(NK)and activated NK cells of patients with HBV infection including chronic hepatitis B(CHB),liver cirrhotic patients,acute hepatitis B patients and healthy individuals;To examine HLA-Cw genotyping in those patients.Moreover to study the relationship between HLA-Cw alleles and the changes of NK cells.Methods(1)To collect peripheral blood samples:111 HBV infection patients(including 30 acute hepatitis B in nonage,30 liver cirrhosis and 51 patients with chronic hepatitis B) and 41 healthy individuals who were enrolled as controls.All patients and controls are Han Chinese from Shandong province.(2)29 patients who were chosen from 51 patients with chronic hepatitis B received IFN-αtherapy for 12 weeks。We collect peripheral blood samples before or after treatment.(3)The numbers of circulating NK cells and activated NK cells were analyzed by flow cytometry.(4)HLA-Cw genotyping was conducted with polymerase chain reaction sequence specific oligonucleotide(PCR-SSO). Results(1)The numbers of circulating NK cells of liver cirrhotic patients,CHB,healthy individuals and acute hepatitis B patients is 13.22%±4.61%,14.36%±1.07%,19.03%±4.91%,22.62%±3.70%,respectively.(2)The numbers of circulating activated NK cells of liver cirrhotic patients,CHB,healthy individuals and acute hepatitis B patients is 45.68%±14.64%, 47.14%±5.67%,53.88%±14.01%,65.28%±14.45%,respectively.(3)The numbers of NK cells and activated NK cells in acute hepatitis B are higher than that in healthy subjects(P＜0.05),The numbers of NK cells and activated NK cells of liver cirrhosis and chronic hepatitis B patients are lower than that in healthy subjects(P＜0.05),there were statistically significant differences between the group of patients with acute hepatitis B and the group of liver cirrhosis,the group of patients with acute hepatitis B and chronic hepatitis B(P＜0.01).(4)The numbers of NK cells and activated NK cells in chronic hepatitis B patients received IFN—αtherapy for 12 weeks were 15.77%±4.25%,46.98%±10.57%, respectively.No significant changes occurred in NK cells compared with those before therapy(P＞0.05).(5)HLA-Cw~*15 was frequent genotypes in the patients with cirrhosis than the healthy(P＜0.05),HLA-Cw~*3 was frequent genotypes in the patients with chronic hepatitis B than the healthy(P＜0.05).No statistically significance was found between the group of acute hepatitis B and healthy subjects about HLA-Cw(P＞0.05).(6)There was a significant negative correlation between the frequency of HLA-Cw~*15and the numbers of activated NK cell in liver cirrhosis(γ=-0.862,P＜0.05).there was a significant negative correlation between the frequency of HLA-Cw~*03 and the numbers of activated NK cell in the group of chronic hepatitis B (γ=-0.394,P＜0.05).Conclusions(1)The numbers and function of NK cells in liver cirrhotic and chronic hepatitis B patients is low,which may be one of the important causes of inducing the lower immunological response of antiviral.(2)The function of NK cells in acute hepatitis B patients is high,which may conduct in cleating HBV.(3)There was no statistically significance in the numbers of NK cells and activated NK cells of chronic hepatitis B patients received IFN—αtherapy for 12 weeks compared with those before therapy,it may be related to short time of IFN—αantiviral therapy.(4)There may be a significant correlation between the frequency of HLA-Cw~* 15 and liver cirrhosis,and induce the damage of liver by effecting the antiviral function of NK cells.(5)As to chronic hepatitis B patients,HLA-Cw~*03 gene may be one of the causes of effecting the antiviral function of NK cells to induce the persistence of HBV infection.