CAJ | 학술논문

目的研究慢性乙型肝炎（CHB）患者外周血来源的树突状细胞（DC）和细胞因子诱导的杀伤细胞（CIK）自体回输治疗的疗效和对机体免疫的影响。方法95例CHB患者随机分为3组，即A组（33例），行DC/CIK治疗；B组（22例），行核苷酸类抗病毒治疗；C组（20例），未进行抗病毒治疗。用CHB外周血培养和扩增DC和CIK，成熟前给予HBsAg刺激。自体静脉回输（DC/CIK细胞数量109/100 ml），1次/3 d，共3次。各组均于治疗前、治疗后第1、3、6、12个月分别检测肝功能、HBV DNA、IFN-γ、IL-4和IL-12水平。其中IFN-γ、IL-4和IL-12均采用双抗体夹心ELISA法进行检测。结果治疗前各组患者病毒载量无显著差异。治疗各阶段B组患者血清HBV DNA水平均低于A组和C组患者（P均＜0.001)；A组患者治疗后3、6个月时病毒载量显著下降（P均＜0.05）。各组患者治疗前后ALT和TBil水平差异无统计学意义。治疗后1个月后，A组患者血清IL-12水平显著高于B组和C组（9.32±2.07 pg/ml vs 6.96±2.17 pg/ml，9.32±2.07 pg/ml vs 7.32±2.36 pg/ml，P＜0.05），其他时间阶段各组比较，IL-12水平差异无统计学意义。各组患者之间血清IFN-γ、IL-4水平差异均无统计学意义。结论 DC/CIK细胞治疗对HBV有一定的抑制作用；推测DC/CIK细胞治疗对辅助T细胞的失衡无显著影响。
목적연구만성을형간염（CHB）환자외주혈래원적수돌상세포（DC）화세포인자유도적살상세포（CIK）자체회수치료적료효화대궤체면역적영향。방법95례CHB환자수궤분위3조，즉A조（33례），행DC/CIK치료；B조（22례），행핵감산류항병독치료；C조（20례），미진행항병독치료。용CHB외주혈배양화확증DC화CIK，성숙전급여HBsAg자격。자체정맥회수（DC/CIK세포수량109/100 ml），1차/3 d，공3차。각조균우치료전、치료후제1、3、6、12개월분별검측간공능、HBV DNA、IFN-γ、IL-4화IL-12수평。기중IFN-γ、IL-4화IL-12균채용쌍항체협심ELISA법진행검측。결과치료전각조환자병독재량무현저차이。치료각계단B조환자혈청HBV DNA수평균저우A조화C조환자（P균＜0.001)；A조환자치료후3、6개월시병독재량현저하강（P균＜0.05）。각조환자치료전후ALT화TBil수평차이무통계학의의。치료후1개월후，A조환자혈청IL-12수평현저고우B조화C조（9.32±2.07 pg/ml vs 6.96±2.17 pg/ml，9.32±2.07 pg/ml vs 7.32±2.36 pg/ml，P＜0.05），기타시간계단각조비교，IL-12수평차이무통계학의의。각조환자지간혈청IFN-γ、IL-4수평차이균무통계학의의。결론 DC/CIK세포치료대HBV유일정적억제작용；추측DC/CIK세포치료대보조T세포적실형무현저영향。
Objective To evaluate the efifcacy and changes of cytokine levels of transfusion of autologous dendritic cells (DC) and cytokine-induced killer cells (CIK) , which were activated by HBsAg for patients with chronic hepatitis B (CHB). Methods Total of 95 CHB patients were selected and divided into group A (n=33, treated with DC/CIK), group B (n=22, nucleotide antiviral therapy) and group C (n=20, with no antiviral treatment). DC and CIK cells were cultured and ampliifed from peripheral blood of CHB patients. DC was stimulated with pure HBsAg in cell culture medium prior to maturation. And 100-150 ml normal saline containing DC/CIK (109) and serum albumin (2%) were infused over 50-60 minutes through a forearm vein, once every three days, totally three times. HBV DNA, biochemical indicators of liver (ALT, TBil) and cytokines (IFN-γ, IL-4 and IL-12) levels in patients were measured before and after 1, 3, 6 and 12 months of treatment. ELISA was applied to detect the levels of IFN-γ, IL-4 and IL-12. Results After all stages of treatment, the HBV DNA load in group B were signiifcantly lower than those in group A and group C (P<0.05). After treatment for 3 and 6 months, the HBV DNA load in group A signiifcantly decreased (P<0.05). The levels of ALT and TBil showed no signiifcant difference among group A, B and C before and after treatment. The serum IL-12 level in group A was higher than that of group B and C (9.32 ± 2.07 pg/ml vs 6.96 ± 2.17 pg/ml, 9.32 ± 2.07 pg/ml vs 7.32 ± 2.36 pg/ml;P<0.05) at a month after treatment, which was not statistically signiifcant in other phases. The levels of IFN-γ, IL-4 were not signiifcantly different among group A, B and C at each stage. Conclusions Transfusion of autologous DC/CIK cells activated by HBsAg had some effects of inhibition of viral replication to patients with CHB. It is suggested that DC/CIK treatment did not improve Th1/Th2 abnormal status in patients with CHB.