Abstract
Caspase activation and degradation of deoxyribonucleic acid (DNA) damage response factors occur during in vitro T-cell proliferation, and an increased frequency of hypoxanthine-guanine phosphoribosyltransferase (HPRT)-negative variants have been reported in conditions associated with in vivo T-cell proliferation. We have applied two human somatic cell mutation reporter assays, for the HPRT and phosphatidylinositol glycan class A (PIG-A) genes, to human T cells activated in vitro with anti-CD3 and anti-CD28. We demonstrate proliferation throughout 6 weeks of cultivation, and find that the frequency of variant cells phenotypically negative for HPRT and PIG-A, respectively, increases from 10−5 up to 10−3–10−2. We also report preliminary evidence for low-density CpG methylation in the HPRT promoter suggesting that epigenetic modification may contribute to this markedly heightened rate of gene inactivation.
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Acknowledgements
We thank Walter Bodmer for generously providing laboratory facilities for the DNA methylation experiments and for the statistic analyses; Kristian Riesbeck for advice on T-cell activation protocol and Johan Bredberg for the mathematical formula. This work was supported by the Swedish Institute, the Swedish Cancer Society, ICRETT Fellowship from UICC Geneva, UMAS Hospital Cancer Foundation, Gunnar Nilsson Cancer Fund and the Greta och Johan Kock, Crafoord, Osterlund and King Gustav V:s 80 year funds.
The experiments performed comply with the current laws of the countries, i.e. Sweden and United Kingdom, in which the experiments were performed.
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Gabdoulkhakova, A., Henriksson, G., Avkhacheva, N. et al. High rate of mutation reporter gene inactivation during human T cell proliferation. Immunogenetics 59, 135–143 (2007). https://doi.org/10.1007/s00251-006-0180-8
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DOI: https://doi.org/10.1007/s00251-006-0180-8