Li Jincheng, Tan Deming, Liu Hongbo, Li Kecheng
Department of Infectious Diseases, Xiangya Hospital, Central South University, Changsha 410008, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2011 Nov;36(11):1046-51. doi: 10.3969/j.issn.1672-7347.2011.11.003.
To determine CD4(+) CD25(+) T regulatory cells (Tregs), forkhead box P3 (FoxP3) mRNA expression and levels of cytokines secreted by peripheral blood mononuclear cells (PBMCs) in individuals responsive or unresponsive to hepatitis B (HB) vaccination, and to explore the relationships between immune response and immune regulatory cells or cytokines.
Based on the antibody against hepatitis B surface antigen (HBsAg) after HB vaccination, the CD4(+) CD25(+) Tregs frequencies in PBMCs from 18 responders, 22 nonresponders and 10 non-immunized healthy controls were analyzed by flow cytometry. The expression of FoxP3 mRNA in PBMCs with or without stimulation of phytohemagglutinin (PHA)and HBsAg was analyzed by real-time quantitative PCR. Levels of IL-4, IL-12, IL-18, and IFN-γ secreted by PBMCs after PHA and HBsAg stimulation were analyzed by enzyme-linked immunosorbent assay.
The ratio of CD4(+) CD25(+) Tregs to CD4(+) T cells in the nonresponders was markedly higher than that in the responders (P<0.05), but lower than that in the controls (P<0.01). FoxP3 was differentially expressed among the responders, nonresponders, and controls in PBMCs before and after PHA and HBsAg stimulation, and nonresponders had the highest FoxP3 mRNA expression (P<0.05 or P<0.01). The content of IFN-γ by PBMCs after PHA and HBsAg stimulation was markedly lower in the nonresponders as compared with the controls and responders (P<0.05). However, there were no significant differences in the levels of IL-18, IL-4, and IL-12 from PBMCs after PHA and HBsAg stimulation between the responders and controls as well as the nonresponders (P>0.05).
CD4(+) CD25(+) FoxP3(+) Tregs may be involved in the negative regulation of responses to hepatitis B vaccination. Immunologic non-responses to hepatitis B vaccination may be related to IFN-γ hyposecretion in PBMCs.
测定对乙型肝炎（HB）疫苗有反应或无反应个体外周血单个核细胞（PBMC）中CD4(+) CD25(+)调节性T细胞（Tregs）、叉头框P3（FoxP3）mRNA表达及细胞因子分泌水平，探讨免疫应答与免疫调节细胞或细胞因子之间的关系。
根据HB疫苗接种后抗乙型肝炎表面抗原（HBsAg）抗体情况，采用流式细胞术分析18名有反应者、22名无反应者及10名未免疫健康对照者PBMC中CD4(+) CD25(+) Tregs频率。采用实时定量PCR分析经植物血凝素（PHA）和HBsAg刺激或未刺激的PBMC中FoxP3 mRNA表达。采用酶联免疫吸附测定法分析PHA和HBsAg刺激后PBMC分泌的IL-4、IL-12、IL-18和IFN-γ水平。
无反应者中CD4(+) CD25(+) Tregs与CD4(+) T细胞的比例显著高于有反应者（P<0.05），但低于对照者（P<0.01）。PHA和HBsAg刺激前后，PBMC中FoxP3在有反应者、无反应者和对照者之间差异表达，无反应者FoxP3 mRNA表达最高（P<0.05或P<0.01）。与对照者和有反应者相比，PHA和HBsAg刺激后无反应者PBMC分泌的IFN-γ含量显著降低（P<0.05）。然而，PHA和HBsAg刺激后，有反应者与对照者以及无反应者PBMC分泌的IL-18、IL-4和IL-12水平无显著差异（P>0.05）。
CD4(+) CD25(+) FoxP3(+) Tregs可能参与HB疫苗接种反应的负调节。对HB疫苗接种的免疫无反应可能与PBMC中IFN-γ分泌不足有关。
