结核菌素纯蛋白衍生物刺激人γδT细胞效应分析

黄方; 杜先智

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结核菌素纯蛋白衍生物刺激人γδT细胞效应分析

论著 | 更新时间：2025-01-15

- 结核菌素纯蛋白衍生物刺激人γδT细胞效应分析
- Activation effect of tuberculin pure protein derivative on γδT cell
- 陆军军医大学学报 2011年33卷第5期页码：481-484
- 作者机构：
  
  重庆医科大学附属第二医院呼吸内科
- 作者简介：
- 基金信息：
- DOI：
  中图分类号： R378.911;R392.7;R392.11
- 纸质出版日期：2011，
  
  网络出版日期：2011-03-15，
- 稿件说明：
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黄方, 杜先智. 结核菌素纯蛋白衍生物刺激人γδT细胞效应分析[J]. 陆军军医大学学报, 2011,33(5):481-484.

HANG FANG, DU XIANZHI. Activation effect of tuberculin pure protein derivative on γδT cell. [J]. Journal of army medical university, 2011, 33(5): 481-484.
黄方, 杜先智. 结核菌素纯蛋白衍生物刺激人γδT细胞效应分析[J]. 陆军军医大学学报, 2011,33(5):481-484. DOI：

HANG FANG, DU XIANZHI. Activation effect of tuberculin pure protein derivative on γδT cell. [J]. Journal of army medical university, 2011, 33(5): 481-484. DOI：

摘要

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(

=20)：①空白组：结核分枝杆菌分泌性抗原（Mtb-Sag）25 μg/ml+CFSE标记的T淋巴细胞；②卡介苗（BCG）组：BCG刺激γδT细胞+Mtb-Sag 25 μg/ml+CFSE标记T淋巴细胞；③PPD组：PPD刺激γδT细胞 + Mtb-Sag 25 μg/ml+CFSE标记T淋巴细胞； ④树突状细胞（dendritic cell

DC)组：成熟DC+Mtb-Sag 25 μg/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）%（

0.05），与BCG刺激效应相似[（42.12±1.72）%，（54.67±2.01）%，（49.79±1.61）%，

>0.05

]

。PPD组T细胞总数（5.14±0.68）×10

和CD4+T细胞所占比例（35.01%）较空白组显著升高（

0.05），与BCG组[（4.6±0.77）×10

，32.00%）

]

相似，显著低于成熟DC细胞组[（7.50±0.35）×10

，58.94%

]

（

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。

Abstract

Objective To explore the activation effect of tuberculin pure protein derivative (PPD) on γδT cells of human peripheral blood. Methods Human peripheral blood mononu

clear cells were separated with Ficoll density gradient centrifugation method

and T cells were separated with a nylon fiber column. After separately stimulated with bacillus calmette-guerin (BCG) and PPD

the γδT cells were sorted and purified with a flow cytometer

and their antigen-presenting cell phenotypes were detected. According to culture medium components of T cells

four experimental groups were established

i.e.

(1) blank group:

Mycobacterium tuberculosis

 (Mtb) secretory antigen (Mtb-Sag) 25 μg/ml+CFSE-labeled T cells

(2) BCG group: BCG-stimulated γδT cells + Mtb-Sag 25 μg/ml+CFSE-labeled T cells

(3) PPD group: PPD-stimulated γδT cells+Mtb-Sag 25 μg/ml+CFSE-labeled T cells

and (4) dendritic cell (DC) group: mature DC+Mtb-Sag 25 μg/ml+CFSE-labeled T cells. The proliferation of T cells and CD4-positive T cells was detected. Results Compared with the resting phase

PPD could stimulate the expression of CD80

CD86 and HLA-DR of γδT cells to reach levels of 39.07±1.26%

51.93±1.85% and 45.74±1.58% (

0.05)

respectively

demonstrating effects comparable to BCG stimulation [(42.12±1.72)%

(54.67±2.01)% and (49.79±1.61)%

>0.05

respectively

]

. The T cell number [(5.14±0.68)×10

]

and the CD4-positive T cell ratio (35.01%) in the PPD group were remarkably higher than those in the blank group [(1.22±0.02)×10

8.28%

0.05

]

similar to those in the BCG group [(4.6±0.77)×10

32.00%

]

and remarkably lower than those in the DC group [(7.50±0.35)×10

58.94%

0.05

]

. Conclusion PPD can stimulate the expression of CD80

CD86 and HLA-DR of γδT cells

indicating its activation effects on human peripheral blood γδT cells.

关键词

结核菌素纯蛋白衍生物γδT细胞流式细胞术

Keywords

tuberculin pure protein derivativeγδT cellflow cytometry

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