title: Measurement of proliferation and disappearance of rapid turnover cell populations in human studies using deuterium-labeled glucose.
abstract: Cell proliferation may be measured in vivo by quantifying DNA synthesis with isotopically labeled deoxyribonucleotide precursors. Deuterium-labeled glucose is one such precursor which, because it achieves high levels of enrichment for a short period, is well suited to the study of rapidly dividing cells, in contrast to the longer term labeling achieved with heavy water ((2)H(2)O). As deuterium is non-radioactive and glucose can be readily administered, this approach is suitable for clinical studies. It has been widely applied to investigate human lymphocyte proliferation, but solid tissue samples may also be analyzed. Rate, duration and route (intravenous or oral) of [6,6-(2)H(2)]-glucose administration should be adapted to the target cell of interest. For lymphocytes, cell separation is best achieved by fluorescence activated cell sorting (FACS), although magnetic bead separation is an alternative. DNA is then extracted, hydrolyzed enzymatically and analyzed by gas chromatography mass spectrometry (GC/MS). Appropriate mathematical modeling is critical to interpretation. Typical time requirements are as follows: labeling, 10-24 h; sampling, approximately 3 weeks; DNA extraction/derivatization, 2-3 d; and GC/MS analysis, approximately 2 d.
authors.affiliation: Centre for Infection, Cellular and Molecular Medicine, St George's, University of London, London, UK. macallan@sgul.ac.uk
authors.forename: Derek C
authors.initials: DC
authors.lastname: Macallan
authors.forename: Becca
authors.initials: B
authors.lastname: Asquith
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Catherine
authors.initials: C
authors.lastname: de Lara
authors.forename: Hala
authors.initials: H
authors.lastname: Ghattas
authors.forename: Julien
authors.initials: J
authors.lastname: Defoiche
authors.forename: Peter C L
authors.initials: PC
authors.lastname: Beverley
mesh_terms: D049109:Cell Proliferation; D004247:DNA / Q000032:analysis / Q000096:biosynthesis; D003903:Deuterium / Q000032:analysis*; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D007553:Isotope Labeling / Q000379:methods*; D008214:Lymphocytes / Q000166:cytology* / Q000378:metabolism*; D008954:Models, Biological; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D004247:DNA; D003903:Deuterium; D005947:Glucose
references.citation: J Immunol Methods. 2003 Aug;279(1-2):199-207
references.pmid: 12969561
references.citation: Trends Immunol. 2002 Dec;23(12):596-601
references.pmid: 12464572
references.citation: Blood. 2005 May 1;105(9):3633-40
references.pmid: 15644412
references.citation: J Immunol. 2008 Jun 15;180(12):7907-18
references.pmid: 18523254
references.citation: Nature. 1992 Nov 19;360(6401):264-5
references.pmid: 1436108
references.citation: J Exp Med. 2007 Sep 3;204(9):2171-85
references.pmid: 17724130
references.citation: Proc Natl Acad Sci U S A. 1998 Jan 20;95(2):708-13
references.pmid: 9435257
references.citation: Nat Med. 2002 Jun;8(6):537-8
references.pmid: 12042783
references.citation: Proc Natl Acad Sci U S A. 2008 Apr 22;105(16):6115-20
references.pmid: 18420820
references.citation: J Exp Med. 2004 Jul 19;200(2):255-60
references.pmid: 15249595
references.citation: J Immunol. 2004 Jul 15;173(2):1209-18
references.pmid: 15240712
references.citation: J Immunol Methods. 1986 Jun 10;90(1):65-70
references.pmid: 3711671
references.citation: Annu Rev Biochem. 1988;57:349-74
references.pmid: 3052277
references.citation: J Infect Dis. 2008 Sep 15;198(6):843-50
references.pmid: 18684102
references.citation: Eur J Immunol. 2003 Aug;33(8):2316-26
references.pmid: 12884307
references.citation: Nat Med. 1999 Jan;5(1):83-9
references.pmid: 9883844
references.citation: J Biol Chem. 2007 Mar 2;282(9):6106-15
references.pmid: 17197446
references.citation: J Virol. 2006 Mar;80(6):2665-74
references.pmid: 16501076
references.citation: J Biol Chem. 1983 Oct 25;258(20):12334-40
references.pmid: 6605343
references.citation: J Immunol Methods. 2000 Sep 21;243(1-2):147-54
references.pmid: 10986412
references.citation: Nature. 1995 Jan 12;373(6510):123-6
references.pmid: 7816094
references.citation: Proc Natl Acad Sci U S A. 2002 Nov 26;99(24):15572-7
references.pmid: 12434018
references.citation: J Exp Med. 2001 Nov 5;194(9):1277-87
references.pmid: 11696593
references.citation: Eur J Immunol. 2003 Oct;33(10):2655-65
references.pmid: 14515249
references.citation: J Immunol. 2007 Jul 1;179(1):36-40
references.pmid: 17579018
references.citation: Hum Immunol. 1991 Feb;30(2):99-104
references.pmid: 1708753
references.citation: J Exp Med. 2001 Dec 17;194(12):1731-41
references.pmid: 11748275
references.citation: J Clin Invest. 2000 Mar;105(5):R1-8
references.pmid: 10712441
references.citation: J Clin Invest. 2003 Sep;112(6):956-66
references.pmid: 12975480
references.citation: Am J Clin Nutr. 2007 Feb;85(2):480-7
references.pmid: 17284747
references.citation: Proc Natl Acad Sci U S A. 2001 May 22;98(11):6313-8
references.pmid: 11344265
references.citation: Nat Protoc. 2007;2(12):3045-57
references.pmid: 18079703
references.citation: Trends Immunol. 2006 Mar;27(3):146-53
references.pmid: 16423560
references.citation: Trans R Soc Trop Med Hyg. 2005 Sep;99(9):675-85
references.pmid: 15985275
references.citation: Immunology. 2007 Jun;121(2):258-65
references.pmid: 17346281
references.citation: J Exp Med. 1994 Apr 1;179(4):1127-35
references.pmid: 8145034
references.citation: Nat Protoc. 2007;2(9):2057-67
references.pmid: 17853861
references.citation: Anal Chem. 2003 Dec 1;75(23):6517-22
references.pmid: 14640722
references.citation: Nat Protoc. 2007;2(9):2049-56
references.pmid: 17853860
references.citation: J Clin Invest. 2005 Aug;115(8):2139-48
references.pmid: 16025158
references.citation: J Leukoc Biol. 2002 Aug;72(2):271-8
references.pmid: 12149417
references.citation: Proc Natl Acad Sci U S A. 2002 Nov 26;99(24):15345-50
references.pmid: 12424339
references.citation: Br J Haematol. 2008 Oct;143(2):240-7
references.pmid: 18710389
references.citation: Immunol Rev. 2007 Apr;216:35-47
references.pmid: 17367333
references.citation: Anal Biochem. 2001 Nov 15;298(2):189-95
references.pmid: 11700973
references.citation: J Immunol. 2004 Aug 1;173(3):1787-94
references.pmid: 15265909
references.citation: Proc Natl Acad Sci U S A. 2007 May 8;104(19):8035-40
references.pmid: 17483473
references.citation: J Clin Invest. 2006 Sep;116(9):2423-33
references.pmid: 16955142
references.citation: Immunol Today. 1999 Oct;20(10):438-41
references.pmid: 10500289