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Background: The quantification of gene expression is an important tool in the evaluation of the immune response to vaccines. Reliable reference genes for gene expression studies in mouse draining lymph nodes after vaccination have not been reported.

Results: The utility of seven potential reference genes was investigated using commercially available Taq-man primer/probe mixes. Results were evaluated with RefFinder, a web-based program including multiple algorithm methods such as geNorm, NormFinder, BestKeeper and the comparative delta-Ct. Further assessment was done by applying the candidate reference genes in relative expression calculations with genes related to the magnitude and longevity of the humoral immune responses. The ubiquitin C gene, Ubc, was found to be the most reliable reference gene when validated with well-known genes that are expressed at relatively low levels after vaccination. The optimal time of sample collection varied depending on the function of the target genes.

Conclusions: This study identified Ubc as the most reliable reference gene and provides useful information for studies examining immunological gene expression in the draining lymph nodes after vaccination in mice.

Titel:	Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination.
Autor/in / Beteiligte Person:	Mosley, YC ; HogenEsch, H
Link:	Full Text Finder (Volltext)
Zeitschrift:	BMC research notes, Jg. 10 (2017-12-06), Heft 1, S. 689
Veröffentlichung:	London : Biomed Central, 2008., 2017
Medientyp:	academicJournal
ISSN:	1756-0500 (electronic)
DOI:	10.1186/s13104-017-3005-y
Schlagwort:	Animals Female Mice Mice, Inbred BALB C Real-Time Polymerase Chain Reaction Gene Expression Lymph Nodes metabolism Vaccines administration & dosage
Sonstiges:	Nachgewiesen in: MEDLINE Sprachen: English Publication Type: Journal Article Language: English [BMC Res Notes] 2017 Dec 06; Vol. 10 (1), pp. 689. <i>Date of Electronic Publication: </i>2017 Dec 06. MeSH Terms: Gene Expression* ; Lymph Nodes / metabolism ; Vaccines / administration & dosage ; Animals ; Female ; Mice ; Mice, Inbred BALB C ; Real-Time Polymerase Chain Reaction References: J Biotechnol. 1999 Oct 8;75(2-3):291-5. (PMID: 10617337) ; Methods. 2001 Dec;25(4):402-8. (PMID: 11846609) ; Vaccine. 2002 May 31;20 Suppl 3:S34-9. (PMID: 12184362) ; Genome Biol. 2002 Jun 18;3(7):RESEARCH0034. (PMID: 12184808) ; Biotechnol Lett. 2004 Mar;26(6):509-15. (PMID: 15127793) ; Biotechniques. 2004 Jul;37(1):112-4, 116, 118-9. (PMID: 15283208) ; Cancer Res. 2004 Aug 1;64(15):5245-50. (PMID: 15289330) ; J Immunol Methods. 2006 May 30;312(1-2):12-9. (PMID: 16624319) ; BMC Mol Biol. 2006 Oct 06;7:33. (PMID: 17026756) ; Biotechniques. 2008 Apr;44(5):619-26. (PMID: 18474036) ; Nat Immunol. 2009 Jan;10(1):116-125. (PMID: 19029902) ; Clin Chem. 2009 Apr;55(4):611-22. (PMID: 19246619) ; J Biochem. 2010 Jun;147(6):793-8. (PMID: 20418328) ; BMC Genomics. 2011 Mar 21;12:156. (PMID: 21418615) ; Nat Immunol. 2011 Jul 10;12(8):786-95. (PMID: 21743478) ; BMC Res Notes. 2011 Oct 20;4:427. (PMID: 22011438) ; J Immunol. 2012 Feb 15;188(4):1751-60. (PMID: 22262658) ; Plant Mol Biol. 2012 Jan 31;:null. (PMID: 22290409) ; J Immunol Methods. 2012 Oct 31;384(1-2):196-9. (PMID: 22884776) ; Crit Rev Immunol. 2012;32(4):287-305. (PMID: 23237506) ; PLoS One. 2013;8(2):e56296. (PMID: 23451040) ; PLoS Pathog. 2013;9(4):e1003264. (PMID: 23592988) ; J Appl Genet. 2013 Nov;54(4):391-406. (PMID: 24078518) ; Anticancer Res. 2013 Dec;33(12):5243-8. (PMID: 24324056) ; Nat Immunol. 2014 Feb;15(2):195-204. (PMID: 24336226) ; J Exp Med. 2014 May 5;211(5):841-56. (PMID: 24711582) ; J Exp Med. 2014 May 5;211(5):827-40. (PMID: 24711583) ; Asian-Australas J Anim Sci. 2013 Dec;26(12):1665-71. (PMID: 25049756) ; BMC Res Notes. 2014 Sep 25;7:675. (PMID: 25256308) ; Ann Clin Lab Sci. 2015 Winter;45(1):64-72. (PMID: 25696013) ; Cell Res. 2016 Apr;26(4):399-422. (PMID: 27012465) ; Cell Res. 2016 Apr;26(4):457-83. (PMID: 27012466) ; NPJ Vaccines. 2017 Feb 23;2:4. (PMID: 29263865) Contributed Indexing: Keywords: DTaP; Lymph nodes; Mouse; Reference gene; Ubc; qPCR Substance Nomenclature: 0 (Vaccines) Entry Date(s): Date Created: 20171207 Date Completed: 20180730 Latest Revision: 20190118 Update Code: 20231215 PubMed Central ID: PMC5718095

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Selection of a suitable reference gene for quantitative gene expression in mouse lymph nodes after vaccination.