Validation of a reliable set of primer pairs for measuring gene expression by real-time quantitative RT-PCR in equine leukocytes

Veterinary Immunology and Immunopathology

Volumn 131, Issue 1-2, 2009, Pages 65-72

  Figueiredo, M D ^a   Salter, C E ^a   Andrietti, A L P ^a   Vandenplas, M L ^a   Hurley, D J ^a   Moore, J N ^a  

^a UNIVERSITY OF GEORGIA   (United States)

Author keywords

Equine; Gene expression; Monocytes; Real time reverse transcription quantitative PCR

Indexed keywords

BACTERIUM LIPOPOLYSACCHARIDE; BETA DEFENSIN; BETA INTERFERON; CATHELICIDIN; CYCLOOXYGENASE 2; GAMMA INTERFERON; GAMMA INTERFERON INDUCIBLE PROTEIN 10; GELATINASE B; HERMES ANTIGEN; HYALURONIDASE; INTERLEUKIN 1 RECEPTOR BLOCKING AGENT; INTERLEUKIN 10; INTERLEUKIN 12P35; INTERLEUKIN 1BETA; INTERLEUKIN 6; INTERLEUKIN 8; L SELECTIN; MESSENGER RNA; MONOCYTE CHEMOTACTIC PROTEIN 3; PLASMINOGEN ACTIVATOR INHIBITOR 1; PROTEIN MD 2; RANTES; RNA 18S; TETRATRICOPEPTIDE REPEAT PROTEIN; TOLL LIKE RECEPTOR 2; TOLL LIKE RECEPTOR 3; TOLL LIKE RECEPTOR 4; TUMOR NECROSIS FACTOR ALPHA; TUMOR NECROSIS FACTOR RECEPTOR ASSOCIATED FACTOR 1;

ANIMAL CELL; ARTICLE; CELL ACTIVATION; CELL ACTIVITY; CELL ISOLATION; CELL STIMULATION; CONTROLLED STUDY; ESCHERICHIA COLI; GEL ELECTROPHORESIS; GENE AMPLIFICATION; GENE EXPRESSION; GENE EXPRESSION PROFILING; GENETIC ANALYSIS; HORSE; IMMUNE RESPONSE; INCUBATION TIME; INFLAMMATION; INFLAMMATORY CELL; LEUKOCYTE; MONOCYTE; NONHUMAN; NUCLEOTIDE SEQUENCE; PATHOGENESIS; QUANTITATIVE ANALYSIS; RELIABILITY; REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION; VALIDATION STUDY;

ANIMALS; DNA PRIMERS; HORSES; LEUKOCYTES; REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION; RNA, MESSENGER;

EQUIDAE; ESCHERICHIA COLI;

EID: 68549110399     PISSN: 01652427     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.vetimm.2009.03.013     Document Type: Article

References (36)

1. Alexopoulou L., Holt A.C., Medzhitov R., and Flavell R.A. Recognition of double-stranded RNA and activation of NF-kappaB by Toll-like receptor 3. Nature 413 (2001) 732-738
2. Allen C.A., Payne S.L., Harville M., Cohen N., and Russell K.E. Validation of quantitative polymerase chain reaction assays for measuring cytokine expression in equine macrophages. J. Immunol. Methods 328 (2007) 59-69
3. Bas A., Forsberg G., Hammarstrom S., and Hammarstrom M.L. Utility of the housekeeping genes 18S rRNA, beta-actin and glyceraldehyde-3-phosphate-dehydrogenase for normalization in real-time quantitative reverse transcriptase-polymerase chain reaction analysis of gene expression in human T lymphocytes. Scand. J. Immunol. 59 (2004) 566-573
4. Breslauer K.J., Frank R., Blocker H., and Marky L.A. Predicting DNA duplex stability from the base sequence. Proc. Natl. Acad. Sci. U.S.A. 83 (1986) 3746-3750
5. Budhia S., Haring L.F., McConnell I., and Blacklaws B.A. Quantitation of ovine cytokine mRNA by real-time RT-PCR. J. Immunol. Methods 309 (2006) 160-172
6. Bustin S.A. Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J. Mol. Endocrinol. 29 (2002) 23-39
7. Cannon J.G., Nerad J.L., Poutsiaka D.D., and Dinarello C.A. Measuring circulating cytokines. J. Appl. Physiol. 75 (1993) 1897-1902
8. Dheda K., Huggett J.F., Bustin S.A., Johnson M.A., Rook G., and Zumla A. Validation of housekeeping genes for normalizing RNA expression in real-time PCR. Biotechniques 37 (2004) 112-114, 116, 118-119
9. Figueiredo M.D., Vandenplas M.L., Hurley D.J., and Moore J.N. Differential induction of MyD88- and TRIF-dependent pathways in equine monocytes by Toll-like receptor agonists. Vet. Immunol. Immunopathol. 15 (2009) 125-134
10. Fontaine G.L., Belknap J.K., Allen D., Moore J.N., and Kroll D.L. Expression of interleukin-1beta in the digital laminae of horses in the prodromal stage of experimentally induced laminitis. Am. J. Vet. Res. 62 (2001) 714-720
11. Fumuso E., Giguere S., Wade J., Rogan D., Videla-Dorna I., and Bowden R.A. Endometrial IL-1beta, IL-6 and TNF-alpha, mRNA expression in mares resistant or susceptible to post-breeding endometritis. Effects of estrous cycle, artificial insemination and immunomodulation. Vet. Immunol. Immunopathol. 96 (2003) 31-41
12. Giguere S., and Prescott J.F. Quantitation of equine cytokine mRNA expression by reverse transcription-competitive polymerase chain reaction. Vet. Immunol. Immunopathol. 67 (1999) 1-15
13. Goidin D., Mamessier A., Staquet M.J., Schmitt D., and Berthier-Vergnes O. Ribosomal 18S RNA prevails over glyceraldehyde-3-phosphate dehydrogenase and beta-actin genes as internal standard for quantitative comparison of mRNA levels in invasive and noninvasive human melanoma cell subpopulations. Anal. Biochem. 295 (2001) 17-21
14. Gutmann S., Zawatzky R., and Muller M. Characterisation and quantification of equine interferon gamma. Vet. Immunol. Immunopathol. 104 (2005) 105-115
15. Huggett J., Dheda K., Bustin S., and Zumla A. Real-time RT-PCR normalisation; strategies and considerations. Genes Immun. 6 (2005) 279-284
16. Klein D. Quantification using real-time PCR technology: applications and limitations. Trends Mol. Med. 8 (2002) 257-260
17. Leutenegger C.M., von Rechenberg B., Huder J.B., Zlinsky K., Mislin C., Akens M.K., Auer J., and Lutz H. Quantitative real-time PCR for equine cytokine mRNA in nondecalcified bone tissue embedded in methyl methacrylate. Calcif. Tissue Int. 65 (1999) 378-383
18. Lim W.S., Edwards J.F., Boyd N.K., Payne S.L., and Ball J.M. Simultaneous quantitation of equine cytokine mRNAs using a multi-probe ribonuclease protection assay. Vet. Immunol. Immunopathol. 91 (2003) 45-51
19. Livak K.J., and Schmittgen T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25 (2001) 402-408
20. Loftus J.P., Black S.J., Pettigrew A., Abrahamsen E.J., and Belknap J.K. Early laminar events involving endothelial activation in horses with black walnut-induced laminitis. Am. J. Vet. Res. 68 (2007) 1205-1211
21. Mire-Sluis A.R., Page L., and Thorpe R. Quantitative cell line based bioassays for human cytokines. J. Immunol. Methods 187 (1995) 191-199
22. Morris D.D., Moore J.N., Fischer K., and Tarleton R.L. Endotoxin-induced tumor necrosis factor activity production by equine peritoneal macrophages. Circ. Shock 30 (1990) 229-236
23. Morrison T.B., Weis J.J., and Wittwer C.T. Quantification of low-copy transcripts by continuous SYBR Green I monitoring during amplification. Biotechniques 24 (1998) 954-962
24. Murphy B.A., Vick M.M., Sessions D.R., Cook R.F., and Fitzgerald B.P. Acute systemic inflammation transiently synchronizes clock gene expression in equine peripheral blood. Brain Behav. Immun. 21 (2007) 467-476
25. Peinnequin A., Mouret C., Birot O., Alonso A., Mathieu J., Clarencon D., Agay D., Chancerelle Y., and Multon E. Rat pro-inflammatory cytokine and cytokine related mRNA quantification by real-time polymerase chain reaction using SYBR green. BMC Immunol. 5 (2004) 3
26. Roge R., Thorsen J., Torring C., Ozbay A., Moller B.K., and Carstens J. Commonly used reference genes are actively regulated in in vitro stimulated lymphocytes. Scand. J. Immunol. 65 (2007) 202-209
27. Rottman J.B., Tompkins W.A., and Tompkins M.B. A reverse transcription-quantitative competitive polymerase chain reaction (RT-qcPCR) technique to measure cytokine gene expression in domestic mammals. Vet. Pathol. 33 (1996) 242-248
28. Schmittgen T.D., and Zakrajsek B.A. Effect of experimental treatment on housekeeping gene expression: validation by real-time, quantitative RT-PCR. J. Biochem. Biophys. Methods 46 (2000) 69-81
29. Sun W.C., Moore J.N., Hurley D.J., Vandenplas M.L., Linden J., Cao Z., and Murray T.F. Adenosine A2A receptor agonists inhibit lipopolysaccharide-induced production of tumor necrosis factor-alpha by equine monocytes. Vet. Immunol. Immunopathol. 121 (2008) 91-100
30. Swiderski C.E., Klei T.R., and Horohov D.W. Quantitative measurement of equine cytokine mRNA expression by polymerase chain reaction using target-specific standard curves. J. Immunol. Methods 222 (1999) 155-169
31. Tschetter J.R., Blikslager A.T., Little D., Howard R.D., Woody S.L., Beex L.M., and Crisman M.V. Detection of differentially regulated genes in ischaemic equine intestinal mucosa. Equine Vet. J. 37 (2005) 319-324
32. Vick M.M., Adams A.A., Murphy B.A., Sessions D.R., Horohov D.W., Cook R.F., Shelton B.J., and Fitzgerald B.P. Relationships among inflammatory cytokines, obesity, and insulin sensitivity in the horse. J. Anim. Sci. 85 (2007) 1144-1155
33. Wagner B., Hillegas J.M., and Antczak D.F. A monoclonal antibody to equine interleukin-4. Vet. Immunol. Immunopathol. 110 (2006) 363-367
34. Wagner B., Hillegas J.M., Brinker D.R., Horohov D.W., and Antczak D.F. Characterization of monoclonal antibodies to equine interleukin-10 and detection of T regulatory 1 cells in horses. Vet. Immunol. Immunopathol. 122 (2008) 57-64
35. Waguespack R.W., Cochran A., and Belknap J.K. Expression of the cyclooxygenase isoforms in the prodromal stage of black walnut-induced laminitis in horses. Am. J. Vet. Res. 65 (2004) 1724-1729
36. Wong M.L., and Medrano J.F. Real-time PCR for mRNA quantitation. Biotechniques 39 (2005) 75-85