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目的:研究对氯苯酚(pCP)对乙肝疫苗反应性差志愿者外周血分离、诱导的DC细胞对乙肝表面抗原肽(HB-sAg)的负载及抗原提呈能力的影响。方法:用梯密度离心法分别分离10例对乙肝疫苗反应性差志愿者外周血单核细胞,黏附的单核细胞培养液中加入HBsAg、rhGM-CSF、rhIL-4、pCP培养7d诱导成熟的DC细胞(实验组),培养体系中不加人pCP为阴性对照组,不加人pCP及HBsAg为空白对照组,ELISA实验检测各自上清其IL-12水平;培养7d后DC细胞与自身T淋巴细胞共培养3d后收集上清,ELISA实验检测其IFN-γ含量。结果:IL-12的水平在实验组(265.68±16.21)ng/L明显高于阴性对照组(168.76±10.01)ng/L(P<0.05)及空白对照组(87±5.79)ng/L(P<0.05);与自身T淋巴细胞共培养3d后,上清中IFN-γ水平在实验组(773.04±32.73)mg/L也明显高于阴性对照组(573.59±26.11)mg/L(P<0.05)及空白对照组(362.81±24.27)mg/L(P<0.05)。结论:pCP能够有效增强乙肝疫苗反应性差志愿者外周血分离、诱导的DC细胞对HBsAg的负载及抗原提成能力,这种效应也能明显增强成熟的CD细胞对自身T淋巴细胞的刺激力,有望成为乙肝疫苗佐剂提高其临床效率。
OBJECTIVE: To study the effect of pCP on the isolation and induction of hepatitis B surface antigen peptide (HBsAg) from antigen-presenting cells in peripheral blood of poorly responsive volunteers with hepatitis B vaccine. Methods: Ten cases of peripheral blood mononuclear cells (PBMCs) with poor responsiveness to hepatitis B vaccine were isolated by density gradient centrifugation. HBsAg, rhGM-CSF and rhIL-4 were added to the adherent mononuclear cell culture medium. (Experimental group), without adding pCP as negative control group in culture system, without adding pCP and HBsAg as blank control group, detecting the supernatant IL-12 level by ELISA; after cultured for 7 days, DC cells were compared with their own T lymphocytes The cells were co-cultured for 3 days and the supernatant was collected. The ELISA assay was used to detect the IFN-γ content. Results: The level of IL-12 in the experimental group (265.68 ± 16.21) ng / L was significantly higher than that in the negative control group (168.76 ± 10.01) ng / L and the blank control group (87 ± 5.79) ng / L (773.04 ± 32.73) mg / L in the experimental group was also significantly higher than that in the negative control group (573.59 ± 26.11) mg / L (P <0.05) <0.05) and blank control group (362.81 ± 24.27) mg / L (P <0.05). Conclusion: pCP can effectively enhance the hepatitis B vaccine-poor volunteers peripheral blood separation, induced by DC cells to HBsAg load and antigen abilities, this effect can significantly enhance the maturation of CD cells on their own T-lymphocyte stimuli, is expected Become a hepatitis B vaccine adjuvant to improve its clinical efficiency.