不同浓度碘对自身免疫性甲状腺疾病患者外周血单个核细胞凋亡、增殖及活化的影响

The effects of different iodide concentrations on apoptosis, proliferation and activation of human peripheral blood mononuclear cells from patients with autoimmune thyroid diseases in vitro

目的 观察不同浓度碘对自身免疫性甲状腺病 (AITD)患者和正常对照者外周血单个核细胞 (PBMC)凋亡、增殖及活化的影响。方法 体外培养AITD患者和正常对照组的PBMC ,单独应用碘化钾或与植物血凝素 (PHA)共同刺激 72h。应用碘化丙啶 (PI)染色方法检测PBMC凋亡的变化 ;应用PI染色方法和3 H TdR掺入实验测定增殖指数的变化 ;应用三标记免疫荧光染色 ,流式细胞仪测定活化 (CD+ 2 5)的辅助性 (CD+ 4 )和抑制性 (CD+ 8)T细胞百分数的变化。结果 基础状态下 ,AITD患者PBMC发生凋亡的百分比明显低于正常对照组。 5× 10 -5mmol/L到 1× 10 -2 mmol/L浓度的碘化钾均明显抑制AITD患者PBMC发生细胞凋亡 ,以 1× 10 -3 mmol/L抑制的最明显 (P <0 .0 5 )。 5× 10 -5mmol/L到 1× 10 -2 mmol/L浓度的碘明显上调AITD患者PBMC在PHA刺激后的增殖指数及活化的CD+ 4 和CD+ 8T淋巴细胞百分比 (P <0 .0 5 )。结论 对于AITD患者 ,在一定浓度范围内变化的碘以剂量依赖方式促进PBMC的异常增殖和活化 ,并抑制其凋亡。

Objective To observe the effects of different iodine cocentrations in vitro on apoptosis, proliferation and activation of peripheral blood mononuclear cell (PBMC) from patients with autoimmune thyroid diseases (AITD) as well as normal control volunteers. Methods The percentage of PBMC apoptosis after 72 h stimulation with potassium, iodide in the presence or absence of PHA was quantified by flow cytometry with propidium iodine (PI) staining. The proliferation index of PBMC after the activation of iodide and PHA was quantified by PI staining and 3\ H thymidine incorporation. Percentages of activated T helper cells (CD + 25 CD + 4) and activated T suppressor cells (CD + 25 CD + 8) under the stimulation of iodide and PHA were also quantified by flow cytometry with direct three color immunofluorescence staining. Results At basic status, the apoptosis percentage of PBMC from the patients with AITD was significantly lower than that of PBMC from normal controls (P<0.05). A significantly suppressed apoptotic rate of PBMC from AITD was found by iodide in the concentration from 5×10 -5 mmol/L to 1×10 -2 mmol/L either with or without PHA. The strongest suppression was observed at the concentration of 1×10 -3 mmol/L iodide (P<0.05). The apoptotic percentage of normal PBMC was suppressed by iodide at any concentration only in the absence of PHA. Iodide significantly increased the proliferation response of PBMC from AITD cultured with PHA in a dose dependent manner withtheiodideconcentrationsfrom5×10 -5 mmol/L to 1×10 -3 mmol/L (P<0.05). Both the percentage of activated T helper cells and T suppressor cells from PBMC of AITD stimulating by PHA were upregulated at iodide concentrations from 5×10 -5 mmol/L to 1×10 -2 mmol/L (P<0.05). On the contrary, neither the percentage of activated T helper cells nor T suppressor cells of PBMC from normalcontrolsatthestimulationof PHA was changed with additional supplement of iodide. Conclusion These results suggest that iodide suppresses apoptosis of PBMC from the patients with AITD and stimulates their proliferation and activation in dose dependent manner at certain iodide cocnentration range which is relevant to physiological concentration.