粒细胞-巨噬细胞集落刺激因子联合乙肝疫苗对宫内感染HBV携带儿童治疗机制初探

Preliminary study on the mechanism of the combined therapy of Hepatitis B virus carrier children with GMCSF and hepatitis B vaccine

目的从细胞因子水平初步探讨粒细胞-巨噬细胞集落刺激因子(GM?CSF)联合乙肝疫苗对宫内HBV感染携带儿童的疗效机制。方法采用ELISA和半定量逆转录?聚合酶链反应(RT?PCR),对12例GM?CSF联合乙肝疫苗治疗后儿童、6例乙肝免疫球蛋白(HBIG)联合乙肝疫苗治疗后儿童及9例未治疗的宫内感染儿童外周血单个核细胞在PHA+LPS、HBsAg及无刺激物时γ?干扰素、白细胞介素(IL)?4分泌及mRNA表达水平进行检测。结果与对照组比较,GM?CSF联合乙肝疫苗治疗儿童γ?干扰素自发分泌显著增加(P=0.017),HBIG联合乙肝疫苗治疗儿童IL?4mRNA转录显著降低(P=0.002)。结论GM?CSF联合乙肝疫苗治疗宫内感染HBV慢性携带儿童时HBV受抑制可能与γ?干扰素产生增多有关,HBIG联合乙肝疫苗治疗时IL?4转录受抑不影响HBV复制,提示慢性HBV感染治疗应以增强Th1应答为主。

Recombinant granulocyte-macrophage colony-stimulating factor GM-CSF is a cytokine that has been used as adjuvant to enhance the immune response to protein and polypeptide antigens. The combined therapy with GM-CSF plus recombinant hepatitis B vaccine rHBvac in hepatitis B virus carrier children has been demonstrated to inhibit the HBV-DNA replication significantly. Therefore this study is aimed to investigate the mechanism of combined therapy with GM-CSF and rHBvac in hepatitis B virus carrier children. Methods Hepatitis B virus carrier children who were born to HBV carrier mothers and received hepatitis B virus prophylaxis after birth,randomized into 2 groups:GM-CSF group receiving combined therapy with 50 μg to GM-CSF plus 10 μg of rHBvac injected intramuscular at same point;HBIG group receiving 200 IU HBIG and 10 μg rHBvac at different site at a four dose schedule in one month interval. 1 to 4 months after the last time inoculation , interleukin-4 (IL-4) and interferon (IFN) secretion and its mRNA of cultured peripheral blood mononuclear cells with different stimulator were determined with ELISA or RT-PCR in 12 children from GM-CSF group,6 from HBIG group and 9 of untreated patients. Results A marked higher spontaneous secretion of IFN was found in the post-treatment children in GM-CSF group than untreated children (P=0.017). A significant less IL-4 mRNA transcription upon PHA and LPS stimulation was found in HBIG group than untreated children(P=0.002). No differences were noted between these three groups upon hepatitis B surface antigen stimulation. Conclusions The increase of IFN secretion may contribute to the inhibition of HBV replication in HBV carrier children with combined GM-CSF and rHBvac therapy.Although the mRNA transcription of IL-4 was reduced by combined HBIG and rHBvac therapy,it was not related to HBV replication,suggesting that the immunotherapy of HBV carrier children should focus on the enhancement of Th1 response.