Li Li, Qiao Dan, Lao Sui-Hua, Chen Shao-Hua, Li Yang-Qiu, Wu Chang-You
Department of Immunology, Zhongshan School of Medicine, Sun Yat-sen University, Key Laboratory of Tropical Disease Control Research of Ministry of Education, Guangzhou 510080, China.
Zhonghua Jie He He Hu Xi Za Zhi. 2010 Oct;33(10):775-8.
To evaluate cytokine production and expression of γδ T cells within pleural fluid cells (PFCs) from patients with tuberculous pleurisy following bacille calmette guerin (BCG) stimulation.
PFCs were isolated from patients with tuberculous pleurisy, and assessed for cytokine production, cell subpopulation, phenotype and characterization of T cell receptors after stimulation with BCG. The positive PCR products were further labeled with fluorescence and analyzed by genescan technique to determine the CDR3 size and evaluate the clonality of the detectable TCR Vγ and Vδ T cells.
Following stimulation with BCG, the positivity of interferon-γ (IFN-γ)-producing CD(4) T cells and γδ T cells were 0.38% and 5.35%, respectively. Phenotypic analysis indicated that the majority of IFN-γ(+)γδ(+) T cells expressed CD(45RO)(+) (73.5%). In addition, δ(2) T cells produced IFN-γ (11.1%) and TNF-α (25.5%). After expansion with BCG for 3 weeks, cells were harvested and mRNA extracted and RT-PCR conducted to amplify cDNA with 3 primers for Vγ and 8 primers for Vδ. The results indicated that BCG selectively expanded δ(2) T cells with oligoclonal peak in Vδ(2) cells.
BCG induced memory γδ and δ(2) T cells to produce cytokines in PFCs. Genescan analysis showed that Vδ(2) displayed oligoclonality.
评估卡介苗（BCG）刺激后结核性胸膜炎患者胸腔积液细胞（PFCs）中γδT细胞的细胞因子产生及表达情况。
从结核性胸膜炎患者中分离出PFCs，在BCG刺激后评估其细胞因子产生、细胞亚群、表型及T细胞受体特征。对阳性PCR产物进一步用荧光标记，通过基因扫描技术分析，以确定CDR3大小并评估可检测到的TCR Vγ和VδT细胞的克隆性。
BCG刺激后，产生干扰素-γ（IFN-γ）的CD4 T细胞和γδT细胞阳性率分别为0.38%和5.35%。表型分析表明，大多数IFN-γ(+)γδ(+) T细胞表达CD45RO(+)（73.5%）。此外，δ2 T细胞产生IFN-γ（11.1%）和肿瘤坏死因子-α（25.5%）。用BCG扩增3周后，收获细胞，提取mRNA并进行RT-PCR，用3种Vγ引物和8种Vδ引物扩增cDNA。结果表明，BCG选择性地扩增了Vδ2细胞中具有寡克隆峰的δ2 T细胞。
BCG诱导记忆性γδ和δ2 T细胞在PFCs中产生细胞因子。基因扫描分析显示Vδ2呈现寡克隆性。
