Stability of expression of reference genes in porcine peripheral blood mononuclear and dendritic cells

被引：24
作者：
Facci, Marina R. [1 ]
Auray, Gael [1 ]
Meurens, Francois [2 ]
Buchanan, Rachelle [1 ]
van Kessel, Jill [1 ]
Gerdts, Volker [1 ,3 ]
机构：
[1] Univ Saskatchewan, Vaccine & Infect Dis Org, Saskatoon, SK S7N 5E3, Canada
[2] INRA, UR1282, F-37380 Nouzilly, France
[3] Univ Saskatchewan, Western Coll Vet Med, Dept Vet Microbiol, Saskatoon, SK S7N 5B4, Canada
来源：
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY | 2011年 / 141卷 / 1-2期
关键词：
Pig; Reference genes; PBMCs; Lymphocytes; Monocytes; Blood-derived dendritic cells; Monocyte-derived dendritic cells;
D O I：
10.1016/j.vetimm.2011.01.005
中图分类号：
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号：
100102 ;
摘要：
Real-time quantitative PCR (RT-qPCR) is a critical tool used to evaluate changes in gene expression. The precision of this tool is reliant upon the selection of reference genes whose expression remains unaltered in culture conditions and following stimulation. Stably expressed reference genes are used to normalize data so observed changes in expression are not due to artifacts but rather reflect physiological changes. In this study, we examined the expression stability of the porcine genes glyceraldehyde 3-phosphate dehydrogenase (GAPDH), succinate dehydrogenase complex subunit A (SDHA), eukaryotic elongation factor 1 gamma-like protein (eEF1), ribosomal protein 119 (RPL19), beta-actin (ACTB) and ATP synthase mitochondrial F0 complex (ATP5G1) in peripheral blood mononuclear cells (PBMCs), monocytes, monocyte-derived dendritic cells (MoDCs), blood isolated dendritic cells (BDCs) and T cells with or without stimulation with lipolysaccharide (LPS). An M value was used as a measure of gene stability as determined using geNORM software. Recommendations for the use of reference genes include using GAPDH and B-actin in PBMCs: RPL19 and SDHA in T cells; RPL19 and B-actin in monocytes; RPL-19 and SDHA in BDCs: and RPL-19 and ATP5GA in MoDCs. (C) 2011 Elsevier B.V. All rights reserved.
引用
收藏
页码：11 / 15
页数：5
相关论文
共 14 条
  • [1] Porcine dendritic cells generated in vitro:: morphological, phenotypic and functional properties
    Carrasco, CP
    Rigden, RC
    Schaffner, R
    Gerber, H
    Neuhaus, V
    Inumaru, S
    Takamatsu, H
    Bertoni, G
    McCullough, KC
    Summerfield, A
    [J]. IMMUNOLOGY, 2001, 104 (02) : 175 - 184
  • [2] Quantitative simultaneous multiplex real-time PCR for the detection of porcine cytokines
    Duvigneau, JC
    Hartl, RT
    Groiss, S
    Gemeiner, M
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 2005, 306 (1-2) : 16 - 27
  • [3] Development of a new set of reference genes for normalization of real-time RT-PCR data of porcine backfat and Longissimus dorsi muscle, and evaluation with PPARGC1A
    Erkens, Tim
    Van Poucke, Mario
    Vandesompele, Jo
    Goossens, Karen
    Van Zeveren, Alex
    Peelman, Luc J.
    [J]. BMC BIOTECHNOLOGY, 2006, 6 (1)
  • [4] Regulation of hypoxanthine phosphoribosyltransferase, glyceraldehyde-3-phosphate dehydrogenase and β-actin mRNA expression in porcine immune cells and tissues
    Foss, DL
    Baarsch, MJ
    Murtaugh, MP
    [J]. ANIMAL BIOTECHNOLOGY, 1998, 9 (01) : 67 - 78
  • [5] Type-A CpG oligonucleotides activate exclusively porcine natural interferon-producing cells to secrete interferon-α, tumour necrosis factor-α and interleukin-12
    Guzylack-Piriou, L
    Balmelli, C
    McCullough, KC
    Summerfield, A
    [J]. IMMUNOLOGY, 2004, 112 (01) : 28 - 37
  • [6] qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data
    Hellemans, Jan
    Mortier, Geert
    De Paepe, Anne
    Speleman, Frank
    Vandesompele, Jo
    [J]. GENOME BIOLOGY, 2007, 8 (02)
  • [7] Kinetics of dendritic cell activation:: impact on priming of TH1, TH2 and nonpolarized T cells
    Langenkamp, A
    Messi, M
    Lanzavecchia, A
    Sallusto, F
    [J]. NATURE IMMUNOLOGY, 2000, 1 (04) : 311 - 316
  • [8] Development of a macroarray to specifically analyze immunological gene expression in swine
    Ledger, TN
    Pinton, P
    Bourges, D
    Roumi, P
    Salmon, H
    Oswald, IP
    [J]. CLINICAL AND DIAGNOSTIC LABORATORY IMMUNOLOGY, 2004, 11 (04) : 691 - 698
  • [9] Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR
    Nygard, Ann-Britt
    Jorgensen, Claus B.
    Cirera, Susanna
    Fredholm, Merete
    [J]. BMC MOLECULAR BIOLOGY, 2007, 8
  • [10] Park MR, 2008, J MICROBIOL BIOTECHN, V18, P1659
12