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Whole blood vs PBMC: compartmental differences in gene expression profiling exemplified in asthma.
Allergy, Asthma & Clinical Immunology ( IF 2.6 ) Pub Date : 2019-11-21 , DOI: 10.1186/s13223-019-0382-x
Daniel He 1, 2 , Chen Xi Yang 1, 2 , Basak Sahin 1 , Amrit Singh 1, 2 , Casey P Shannon 2 , John-Paul Oliveria 3, 4 , Gail M Gauvreau 3 , Scott J Tebbutt 1, 2, 5
Affiliation  

Background Blood has proven to be a useful resource for molecular analysis in numerous biomedical studies, with peripheral blood mononuclear cells (PBMCs) and whole blood being the major specimen types. However, comparative analyses between these two major compartments (PBMCs and whole blood) are few and far between. In this study, we compared gene expression profiles of PBMCs and whole blood samples obtained from research subjects with or without mild allergic asthma. Methods Whole blood (PAXgene) and PBMC samples were obtained from 5 mild allergic asthmatics and 5 healthy controls. RNA from both sample types was measured for expression of 730 immune-related genes using the NanoString nCounter platform. Results We identified 64 uniquely expressed transcripts in whole blood that reflected a variety of innate, humoral, and adaptive immune processes, and 13 uniquely expressed transcripts in PBMCs which were representative of T-cell and monocyte-mediated processes. Furthermore, analysis of mild allergic asthmatics versus non-asthmatics revealed 47 differentially expressed transcripts in whole blood compared to 1 differentially expressed transcript in PBMCs (FDR < 0.25). Finally, through simultaneous measurement of PBMC proteins on the nCounter assay, we identified CD28 and OX40 (TNFRSF4), both of which are critical co-stimulatory molecules during T-cell activation, as significantly upregulated in asthmatics. Conclusions Whole blood RNA preserved in PAXgene tubes is excellent for producing gene expression data with minimal variability and good sensitivity, suggesting its utility in multi-centre studies requiring measurement of blood gene expression.

中文翻译:

全血与 PBMC:以哮喘为例的基因表达谱的区室差异。

背景 血液已被证明是许多生物医学研究中分子分析的有用资源,外周血单核细胞 (PBMC) 和全血是主要的样本类型。然而,这两个主要部分(PBMC 和全血)之间的比较分析很少而且相差甚远。在这项研究中,我们比较了从患有或不患有轻度过敏性哮喘的研究对象获得的 PBMC 和全血样本的基因表达谱。方法 从 5 名轻度过敏性哮喘患者和 5 名健康对照者中获取全血(PAXgene)和 PBMC 样本。使用 NanoString nCounter 平台测量了两种样品类型的 RNA 中 730 个免疫相关基因的表达。结果 我们在全血中鉴定出 64 个独特表达的转录本,它们反映了各种先天性、体液性、和适应性免疫过程,以及 13 个在 PBMC 中独特表达的转录物,它们代表了 T 细胞和单核细胞介导的过程。此外,对轻度过敏性哮喘患者与非哮喘患者的分析显示,全血中有 47 个差异表达的转录物,而 PBMC 中有 1 个差异表达的转录物(FDR < 0.25)。最后,通过在 nCounter 测定中同时测量 PBMC 蛋白,我们确定了 CD28 和 OX40 (TNFRSF4),这两者都是 T 细胞活化过程中的关键共刺激分子,在哮喘患者中显着上调。结论 保存在 PAXgene 试管中的全血 RNA 非常适合产生变异性最小且灵敏度高的基因表达数据,这表明它在需要测量血液基因表达的多中心研究中的实用性。和 13 个在 PBMC 中独特表达的转录本,它们代表了 T 细胞和单核细胞介导的过程。此外,对轻度过敏性哮喘患者与非哮喘患者的分析显示,全血中有 47 个差异表达的转录物,而 PBMC 中有 1 个差异表达的转录物(FDR < 0.25)。最后,通过在 nCounter 测定中同时测量 PBMC 蛋白,我们确定了 CD28 和 OX40 (TNFRSF4),这两者都是 T 细胞活化过程中的关键共刺激分子,在哮喘患者中显着上调。结论 保存在 PAXgene 试管中的全血 RNA 非常适合产生变异性最小且灵敏度高的基因表达数据,这表明它在需要测量血液基因表达的多中心研究中的实用性。和 13 个在 PBMC 中独特表达的转录本,它们代表了 T 细胞和单核细胞介导的过程。此外,对轻度过敏性哮喘患者与非哮喘患者的分析显示,全血中有 47 个差异表达的转录物,而 PBMC 中有 1 个差异表达的转录物(FDR < 0.25)。最后,通过在 nCounter 测定中同时测量 PBMC 蛋白,我们确定了 CD28 和 OX40 (TNFRSF4),这两者都是 T 细胞活化过程中的关键共刺激分子,在哮喘患者中显着上调。结论 保存在 PAXgene 试管中的全血 RNA 非常适合产生变异性最小且灵敏度高的基因表达数据,这表明它在需要测量血液基因表达的多中心研究中的实用性。
更新日期:2020-04-22
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