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Comparative Study
. 2010 Jan 31;352(1-2):71-80.
doi: 10.1016/j.jim.2009.09.012. Epub 2009 Oct 21.

Analyses and comparisons of telomerase activity and telomere length in human T and B cells: insights for epidemiology of telomere maintenance

Jue Lin  1 , Elissa Epel, Joshua Cheon, Candyce Kroenke, Elizabeth Sinclair, Marty Bigos, Owen Wolkowitz, Synthia Mellon, Elizabeth Blackburn
Affiliations
Comparative Study

Analyses and comparisons of telomerase activity and telomere length in human T and B cells: insights for epidemiology of telomere maintenance

Jue Lin et al. J Immunol Methods. .

Abstract

Telomeres are the DNA-protein complexes that protect the ends of eukaryotic chromosomes. The cellular enzyme telomerase counteracts telomere shortening by adding telomeric DNA. A growing body of literature links shorter telomere length and lower telomerase activity with various age-related diseases and earlier mortality. Thus, leukocyte telomere length (LTL) and telomerase activity are emerging both as biomarkers and contributing factors for age-related diseases. However, no clinical study has directly examined telomerase activity and telomere length in different lymphocyte subtypes isolated from the same donors, which could offer insight into the summary measure of leukocyte telomere maintenance. We report the first quantitative data in humans examining both levels of telomerase activity and telomere length in four lymphocyte subpopulations from the same donors-CD4+, CD8+CD28+ and CD8+CD28- T cells and B cells, as well as total PBMCs-in a cohort of healthy women. We found that B cells had the highest telomerase activity and longest telomere length; CD4+ T cells had slightly higher telomerase activity than CD8+CD28+ T cells, and similar telomere length. Consistent with earlier reports that CD8+CD28- T cells are replicatively senescent cells, they had the lowest telomerase activity and shortest telomere length. In addition, a higher percentage of CD8+CD28- T cells correlated with shorter total PBMC TL (r=-0.26, p=0.05). Interestingly, telomerase activities of CD4+ and CD8+CD28+ T cells from the same individual were strongly correlated (r=0.55, r<0.001), indicating possible common mechanisms for telomerase activity regulation in these two cell subtypes. These data will facilitate the understanding of leukocyte aging and its relationship to human health.

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Figures

Fig. 1
Fig. 1
A typical gel phosphorimager image showing TRAP activity using the TRAPeze kit, from the four cell types from one study subject. +: Standard sample containing extracts from 20 293T cells. 2000, 5000 and 10,000 cells were used for each sample. The arrow points to the non-specific band and the circle points to the internal control band (see Materials and methods).
Fig. 2
Fig. 2
Comparison of telomerase activity with various parameters during blood draw. A) heparin (green top) vs. EDTA (lavender top) collection tubes from the same individual (n = 9). B) blood draw in the morning vs. in the afternoon from the same individual (n = 9). C) Room temperature for 2 h vs. immediate processing of blood from the same individual (n = 9).
Fig. 3
Fig. 3
FACS sorting panel.
Fig. 4
Fig. 4
Box plots for telomerase activity and telomere length.
See this image and copyright information in PMC

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