| Chronic hepatitis B(CHB)is defined as hepatitis B virus(HBV)infection persists for more than 6 months.It is reported that at least 250 million people wordwide are serologic HBsAg positive and there are approximately 20 million CHB patients in China.Current therapies to CHB rarely achieve a clinical cure because those therapies have little effect on the viral replication intermediate named covalently close circular DNA(cccDNA).A clinical cure will therefore require elimination of cccDNA in these liver cells infected HBV.Until now,little is known about the molecular basis of cccDNA formation and degradation due to the lack of suitable experimental model.Enticavir is extensively used for the treatment of adult CHB patients with highly activity of viral replication and/or increased level of alanine aminotransferase(ALT).Histological evaluation after long term treatment with ETV showed an improvement in necroinflammatory and fibrosis scores in most patients.For hepatitis B e antigen positive patients,48 weeks of ETV monotherapy led to HBeAg seroconversion in 21% patients,and approximately 67 percent patients responsed with serum HBV DNA < 400 copies/ml.After 5 years of follow-up,80 percent of patients responsed with serum ALT normalization,and the serum HBV DNA copies decreased to the level cannot be detected in 94% patients.To improve the efficacy and mitigate the side effect of the treatment with ETV,we designed a clinical trial in which the CHB patients treated with ETV with and without granulocyte macrophages colony stimulating factor(GM-CSF).GM-CSF is a cytokine produced by T lymphocytes,natural killer cells(NK cells)and macrophages that can boost the hematopoietic stem cells differentiate to granulocyte and monocytes.GM-CSF also show other biological activity including the augment the activity of granulocytes and NK cells.These patients enrolled in our stduy divided into two groups,and one group patients(group ETV)received a monotherapy with 0.5 mg of enticavir every day,the other group patients(group ETV+GM)received 100 ?g GM-CSF hypodermic injection in the day 3,day4,and day5 in the week 1,week 4,week 12,week 24 and week 48 on the basic treatment with enticacir.Our results showed that the frequencies of CD3-CD56+ CD16+cells in group ETV and group ETV+GM were decreased compared to those in group health control(HC).Before treatment,the frequencies of CD3-CD56+ CD16+cells were 11.86±2.20% and 11.59±5.71%;and those frequencies were 13.85±4.80% and 11.28±4.96%,12.79±3.14% and 12.93±9.7%,11.09±6.95% and 12.47±3.49% at week 12,week 24,week 36 respectively?There were no difference between those groups.In group ETV and group ETV+GM,the frequencies of CD45RA+CD69-NK cells were(29.18±21.76% vs 36.14±22.32%),(35.00±32.44% vs 61.03±10.54%),(51.30±28.32% vs 55.13±29.29%),(87.45±4.2% vs 81.31±2.393%),respectively.After treated with ETV with or without GM-CSF,the frequencies of CD45RA+CD69-NK cells increased with time by treatment.The frequencies of CD45RA+CD69-NK cells in group ETV+GM was significant high than those in group ETV.Western blot depicting a high level of constitutive phospho-mTOR(Ser2448)and phospho-p70 S6 kinase(Thr389 and Ser371)in C D56+ NK cells in CHB patients treated with enticavir with or withou t GM-CSF.The NK cells of CHB patients treated with enticavir no t expressed the phosphor-mTOR(7C10)and phospho-4E-BP1(Thr37/46).The GM-CSF can decreased the expression of the phosphor-p70S6 kinase and phosphor-mTOR(Ser2448),and increased the express ion of the mTOR(7C10)and phosphor-4E-BP1(Thr37/46).In conclusion,the treatment with ETV and GM-CSF can improve the proportion of CD45RA+CD69-cells of NK cells,and augment the protein phosphorylation level of the protein of chronic hepatitis B patients.These results showed the GM-CSF the activation of metabolism of the periphery NK cells of the CHB patients treated with the ETV. |
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