title: Immune-related gene expression profile in laboratory common marmosets assessed by an accurate quantitative real-time PCR using selected reference genes.
abstract: 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.
authors: Department of Rheumatology and Clinical Immunology, Clinical Research Center for Allergy and Rheumatology, Sagamihara National Hospital, National Hospital Organization, Kanagawa, Japan.
authors: Yoshiki
authors: Y
authors: Fujii
authors: Kazutaka
authors: K
authors: Kitaura
authors: Takaji
authors: T
authors: Matsutani
authors: Kenji
authors: K
authors: Shirai
authors: Satsuki
authors: S
authors: Suzuki
authors: Tomohiko
authors: T
authors: Takasaki
authors: Kenichi
authors: K
authors: Kumagai
authors: Yoshie
authors: Y
authors: Kametani
authors: Takashi
authors: T
authors: Shiina
authors: Shuji
authors: S
authors: Takabayashi
authors: Hideki
authors: H
authors: Katoh
authors: Yoshiki
authors: Y
authors: Hamada
authors: Ichiro
authors: I
authors: Kurane
authors: Ryuji
authors: R
authors: Suzuki
mesh_terms: D000818:Animals; D002144:Callithrix / Q000235:genetics*; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics; D016753:Interleukin-10 / Q000235:genetics; D018793:Interleukin-13 / Q000235:genetics; D007376:Interleukin-2 / Q000235:genetics; D015847:Interleukin-4 / Q000235:genetics; D015848:Interleukin-5 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D059467:Transcriptome / Q000235:genetics*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
chemical_list: D018793:Interleukin-13; D007376:Interleukin-2; D015848:Interleukin-5; D015850:Interleukin-6; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D015847:Interleukin-4; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)