Immune-related gene expression profile in laboratory common marmosets assessed by an accurate quantitative real-time PCR using selected reference genes.
Immune-related gene expression profile in laboratory common marmosets assessed by an accurate quantitative real-time PCR using selected reference genes.
复制标题
DOI：
10.1371/journal.pone.0056296
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Suzuki R
中科院分区：
文献类型：
--
作者：
Fujii Y;Kitaura K;Matsutani T;Shirai K;Suzuki S;Takasaki T;Kumagai K;Kametani Y;Shiina T;Takabayashi S;Katoh H;Hamada Y;Kurane I;Suzuki R
普通绒猴（Callithrix jacchus）被认为是一种新型的非人灵长类实验动物模型。然而，由于抗体的不可用，免疫学和病理学研究还没有充分进行各种疾病模型的普通绒猴。定量实时PCR（qPCR）是检测基因表达水平的有力工具。最近的报道表明，选择内参管家基因是基因表达精确标准化所必需的。为了开发一种可靠的普通绒猴qPCR方法，我们使用geNorm applets评估了8个候选参考基因（GAPDH、ACTB、rRNA、B2 M、UBC、HPRT、SDHA和TBP）在实验室普通绒猴不同组织中的表达稳定性。geNorm分析显示GAPDH、ACTB、SDHA和TBP的稳定性普遍较高，其次是UBC。相比之下，HPRT，rRNA和B2 M表现出较低的表达稳定性比其他基因在大多数组织分析。此外，我们还利用改良的qPCR技术，结合所选的参考基因，分析了CD抗原（CD 3 ε、CD 4、CD 8 α和CD 20）和细胞因子（IL-1β、IL-2、IL-4、IL-5、IL-6、IL-10、IL-12β、IL-13、IFN-γ和TNF-α）在外周血白细胞中的表达水平，并比较了普通绒猴和人类的表达水平。普通绒猴CD 4和IL-4的表达水平低于人类，而IL-10、IL-12β和IFN-γ的表达水平高于人类。普通绒猴Th 1/Th 2相关基因的表达水平呈倒置。我们通过流式细胞术分析证实了普通绒猴的CD 4/CD 8倒置。因此，普通绒猴与人类之间Th 1/Th 2平衡的差异可能会影响宿主防御和/或疾病易感性，在使用普通绒猴作为生物医学研究的实验模型时应仔细考虑。
The common marmoset (Callithrix jacchus) is considered a novel experimental animal model of non-human primates. However, due to antibody unavailability, immunological and pathological studies have not been adequately conducted in various disease models of common marmoset. Quantitative real-time PCR (qPCR) is a powerful tool to examine gene expression levels. Recent reports have shown that selection of internal reference housekeeping genes are required for accurate normalization of gene expression. To develop a reliable qPCR method in common marmoset, we used geNorm applets to evaluate the expression stability of eight candidate reference genes (GAPDH, ACTB, rRNA, B2M, UBC, HPRT, SDHA and TBP) in various tissues from laboratory common marmosets. geNorm analysis showed that GAPDH, ACTB, SDHA and TBP were generally ranked high in stability followed by UBC. In contrast, HPRT, rRNA and B2M exhibited lower expression stability than other genes in most tissues analyzed. Furthermore, by using the improved qPCR with selected reference genes, we analyzed the expression levels of CD antigens (CD3ε, CD4, CD8α and CD20) and cytokines (IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12β, IL-13, IFN-γ and TNF-α) in peripheral blood leukocytes and compared them between common marmosets and humans. The expression levels of CD4 and IL-4 were lower in common marmosets than in humans whereas those of IL-10, IL-12β and IFN-γ were higher in the common marmoset. The ratio of Th1-related gene expression level to that of Th2-related genes was inverted in common marmosets. We confirmed the inverted ratio of CD4 to CD8 in common marmosets by flow cytometric analysis. Therefore, the difference in Th1/Th2 balance between common marmosets and humans may affect host defense and/or disease susceptibility, which should be carefully considered when using common marmoset as an experimental model for biomedical research.
登录
查看更多内容
影响因子：
3.4
作者：
Ando, Kiyoshi;Maeda, Jun;Tanioka, Yoshikuni
通讯作者：
Tanioka, Yoshikuni
影响因子：
6.2
作者：
DAAR, AS;FUGGLE, SV;MORRIS, PJ
通讯作者：
MORRIS, PJ
影响因子：
5.4
作者：
Adams, A. Paige;Aronson, Judith F.;Weaver, Scott C.
通讯作者：
Weaver, Scott C.
影响因子：
5.4
作者：
Kimura, T;Griffin, DE
通讯作者：
Griffin, DE
影响因子：
--
作者：
Bostik P;Watkins M;Villinger F;Ansari AA
通讯作者：
Ansari AA