Quantitation of rabbit cytokine mRNA by real-time RT-PCR

被引：103
|
作者：
Godornes, Charmie
Leader, Brandon Troy
Molini, Barbara J.
Centurion-Lara, Arturo
Lukehart, Sheila A.
机构：
[1] Univ Washington, Dept Med, Harborview Med Ctr, Seattle, WA 98104 USA
[2] Univ Washington, Dept Pathobiol, Seattle, WA 98104 USA
来源：
CYTOKINE | 2007年 / 38卷 / 01期
关键词：
rabbit; cytokines; quantitative real-time RT-PCR; syphilis;
D O I：
10.1016/j.cyto.2007.04.002
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Fundamental understanding of rabbit immunology and the use of the rabbit as a disease model have long been hindered by the lack of immunological assays specific to this species. In the present study, we sought to develop a method to quantitate cytokine expression in rabbit cells and tissues. We report the development of a quantitative real-time RT-PCR method for measuring the relative levels of rabbit IFN-gamma IL-2, IL-4 IL-10 and TNF-alpha mRNA. Quantitation was accomplished by comparison to a standard curve generated using plasmid DNA containing partial sequences of the relevant cytokines. Experimental studies demonstrate applicability of this assay to quantitate cytokine mRNA levels from rabbit spleen cells following mitogen stimulation. We have further utilized this assay to also examine cytokine expression in rabbit tissues during experimental syphilis infection. (c) 2007 Elsevier Ltd. All rights reserved.
引用
收藏
页码：1 / 7
页数：7
相关论文
共 50 条
  • [1] Quantitation of ovine cytokine mRNA by real-time RT-PCR
    Budhia, S
    Haring, LF
    McConnell, I
    Blacklaws, BA
    [J]. JOURNAL OF IMMUNOLOGICAL METHODS, 2006, 309 (1-2) : 160 - 172
  • [2] Quantitation of cytokine mRNA by real-time RT-PCR during a vaccination trial in a rabbit model of fascioliasis
    Espino, Ana M.
    Rivera, Francheska
    [J]. VETERINARY PARASITOLOGY, 2010, 169 (1-2) : 82 - 92
  • [3] Quantification of mRNA using real-time RT-PCR
    Nolan, Tania
    Hands, Rebecca E.
    Bustin, Stephen A.
    [J]. NATURE PROTOCOLS, 2006, 1 (03) : 1559 - 1582
  • [4] Quantification of mRNA using real-time RT-PCR
    Tania Nolan
    Rebecca E Hands
    Stephen A Bustin
    [J]. Nature Protocols, 2006, 1 : 1559 - 1582
  • [5] Real-time RT-PCR for the detection and quantitation of Oropouche virus
    Rojas, Alejandra
    Stittleburg, Victoria
    Cardozo, Fatima
    Bopp, Nathen
    Cantero, Cesar
    Lopez, Sanny
    Bernal, Cynthia
    Mendoza, Laura
    Aguilar, Patricia
    Pinsky, Benjamin A.
    Guillen, Yvalena
    Paez, Malvina
    Waggoner, Jesse J.
    [J]. DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE, 2020, 96 (01)
  • [6] Cytokine mRNA quantification in histologically normal canine duodenal mucosa by real-time RT-PCR
    Peters, IR
    Helps, CR
    Calvert, EL
    Hall, EJ
    Day, MJ
    [J]. VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 2005, 103 (1-2) : 101 - 111
  • [7] Real-time RT-PCR for quantitation of hepatitis C virus RNA
    Yang, JH
    Lai, JP
    Douglas, SD
    Metzger, D
    Zhu, XH
    Ho, WZ
    [J]. JOURNAL OF VIROLOGICAL METHODS, 2002, 102 (1-2) : 119 - 128
  • [8] Real-time PCR for mRNA quantitation
    Wong, ML
    Medrano, JF
    [J]. BIOTECHNIQUES, 2005, 39 (01) : 75 - 85
  • [9] Quantification of llama inflammatory cytokine mRNAs by real-time RT-PCR
    Odbileg, R
    Konnai, S
    Usui, T
    Ohashi, K
    Onuma, M
    [J]. JOURNAL OF VETERINARY MEDICAL SCIENCE, 2005, 67 (02): : 195 - 198
  • [10] Validation of real-time RT-PCR assays for mRNA quantification in baboons
    Overbergh, L
    Kyama, CM
    Valckx, D
    Debrock, S
    Mwenda, JM
    Mathieu, C
    D'Hooghe, T
    [J]. CYTOKINE, 2005, 31 (06) : 454 - 458
12345