目的探讨结核菌素纯蛋白衍生物(pure protein derivative,PPD)对人外周血γδT细胞的刺激效应。方法采用Ficoll密度梯度离心法分离人外周血单个核细胞(peripheral blood mononuclear cell,PBMC)后,经尼龙毛柱法分离得T细胞。分别以卡介苗和PPD刺激γδT细胞,流式细胞仪分选纯化γδT细胞,并检测其抗原提呈细胞表型。按照T细胞培养基成分将实验分为4组(n=20):①空白组:结核分枝杆菌分泌性抗原(Mtb-Sag)25μg/ml+CFSE标记的T淋巴细胞;②卡介苗(BCG)组:BCG刺激γδT细胞+Mtb-Sag 25μg/ml+CFSE标记T淋巴细胞;③PPD组:PPD刺激γδT细胞+Mtb-Sag 25μg/ml+CFSE标记T淋巴细胞;④树突状细胞(dendritic cell,DC)组:成熟DC+Mtb-Sag 25μg/ml+CFSE标记T淋巴细胞。9 d后检测T细胞及CD4+T细胞的增殖情况。结果与静止期相比,PPD可刺激γδT细胞CD80、CD86和HLA-DR表达水平升高至(39.07±1.26)%、(51.93±1.85)%和(45.74±1.58)%(P<0.05),与BCG刺激效应相似[(42.12±1.72)%,(54.67±2.01)%,(49.79±1.61)%,P>0.05]。PPD组T细胞总数(5.14±0.68)×106和CD4+T细胞所占比例(35.01%)较空白组显著升高(P<0.05),与BCG组[(4.6±0.77)×106,32.00%)]相似,显著低于成熟DC细胞组[(7.50±0.35)×106,58.94%](P<0.05)。结论 PPD可激活γδT细胞CD80、CD86和HLA-DR表达,提示PPD可能具有激活人外周γδT细胞功能。 Objective To investigate the stimulatory effect of pure protein derivative (PPD) on human peripheral blood γδT cells. Methods Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll density gradient centrifugation. T cells were isolated by nylon hair column method. ΓδT cells were stimulated with BCG and PPD respectively. ΓδT cells were sorted by flow cytometry and their antigen presenting cell phenotypes were detected. According to the components of T cell culture medium, the experiment was divided into 4 groups (n = 20): ① Blank group: 25μg / ml Mtb-Sag + CFSE labeled T lymphocytes; ② BCG (3) PPD group: γδT cells + Mtb-Sag 25μg / ml + CFSE-labeled T lymphocytes stimulated by BCG; ④ dendritic cells DC): Mature DC + Mtb-Sag 25 μg / ml + CFSE-labeled T lymphocytes. After 9 days, the proliferation of T cells and CD4 + T cells were detected. Results PPD stimulated the expression of CD80, CD86 and HLA-DR in γδT cells to (39.07 ± 1.26)%, (51.93 ± 1.85)% and (45.74 ± 1.58)%, respectively Similar to the BCG stimulation effect [(42.12 ± 1.72)%, (54.67 ± 2.01)%, (49.79 ± 1.61)%, P> 0.05]. The total number of T cells (5.14 ± 0.68) × 106 and CD4 + T cells (35.01%) in PPD group was significantly higher than that in blank group ([4.6 ± 0.77] × 106,32.00% )], Which was significantly lower than that in mature DCs [(7.50 ± 0.35) × 106,58.94%] (P <0.05). Conclusion PPD can activate γδT cells CD80, CD86 and HLA-DR expression, suggesting that PPD may have the activation of human peripheral γδ T cell function.