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Quantitation of rabbit cytokine mRNA by real-time RT-PCR.
Title | Quantitation of rabbit cytokine mRNA by real-time RT-PCR. |
Publication Type | Journal Article |
Year of Publication | 2007 |
Authors | Godornes, C, Leader, BTroy, Molini, BJ, Centurion-Lara, A, Lukehart, SA |
Journal | Cytokine |
Volume | 38 |
Issue | 1 |
Pagination | 1-7 |
Date Published | 2007 Apr |
ISSN | 1043-4666 |
Keywords | Animals, Concanavalin A, Cytokines, Male, Plasmids, Rabbits, Reverse Transcriptase Polymerase Chain Reaction, RNA, Messenger, Spleen, Syphilis, Cutaneous |
Abstract | 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. |
DOI | 10.1016/j.cyto.2007.04.002 |
Alternate Journal | Cytokine |
PubMed ID | 17521914 |
PubMed Central ID | PMC2041851 |
Grant List | AI 34616 / AI / NIAID NIH HHS / United States AI42143 / AI / NIAID NIH HHS / United States AI63940 / AI / NIAID NIH HHS / United States P01 AI034616 / AI / NIAID NIH HHS / United States P01 AI034616-090007 / AI / NIAID NIH HHS / United States P01 AI034616-10 / AI / NIAID NIH HHS / United States R01 AI042143 / AI / NIAID NIH HHS / United States R01 AI042143-09A1 / AI / NIAID NIH HHS / United States R01 AI063940 / AI / NIAID NIH HHS / United States R01 AI063940-04 / AI / NIAID NIH HHS / United States |