Freeze-thaw lysates of Plasmodium falciparum-infected red blood cells induce differentiation of functionally competent regulatory T cells from memory T cells
Article first published online: 17 JUL 2012
© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
European Journal of Immunology
Volume 42, Issue 7, pages 1767–1777, July 2012
How to Cite
Finney, O. C., Lawrence, E., Gray, A. P., Njie, M., Riley, E. M. and Walther, M. (2012), Freeze-thaw lysates of Plasmodium falciparum-infected red blood cells induce differentiation of functionally competent regulatory T cells from memory T cells. Eur. J. Immunol., 42: 1767–1777. doi: 10.1002/eji.201142164
- Issue published online: 17 JUL 2012
- Article first published online: 17 JUL 2012
- Accepted manuscript online: 14 MAY 2012 07:41AM EST
- Manuscript Accepted: 11 APR 2012
- Manuscript Revised: 23 DEC 2011
- Manuscript Received: 10 OCT 2011
Disclaimer: Supplementary materials have been peer-reviewed but not copyedited.
|eji2305-sup-0001-figures1.pdf||297K||Suppl. figure 1 Experimental design and cell purities A scheme of the experimental approach is depicted in Figure (A). Viable CD4+ T cells amongst (B): PBMC, (D): ‘Tmem’, (F): ‘Tnaive’ are shown according to their expression of memory markers CD45RA/RO, and according to their expression of Treg markers FOXP3 and CD127 (C: PBMC, E: ‘Tmem’, G: ‘Tnaive’, I: ‘APC’ and K: ‘Treg’). Compared to undepleted PBMC ‘Tmem’ were depleted of > 98% of CD4+ CD45RA+ and > 74% of CD4+ FOXP3+ CD127lo/- T cells. ‘Tnaive’ cells were depleted > 97% of CD4+ CD45RO+ and > 80% of CD4+ FOXP3+ CD127lo/- T cells compared to PBMC. (H): the percentage of viable cells expressing HLA-DR amongst ‘APC’ is shown. The ‘APC’ fraction comprised of a more than 77% pure HLADR+ population, of which > 94% of CD4+ FOXP3+ CD127lo/- T cells were depleted compared to PBMC. (J): Isolated ‘Treg’ were uniformly labeled with CFSE and contained at least 54% CD4+ FOXP3+ CD127lo/- cells (K). Note, for better comparability, percentages depicted in figures B-K refer to the proportion amongst viable cells, not CD4+ cells. The proportion of viable CD4+ T cells expressing a Treg phenotype in the cell mix containing (L): ‘Tnaive’ + ‘APC’ + ‘Treg’, or (N): ‘Tmem’ + ‘APC’ + ‘Treg’ is shown; The CFSE+ cells are depicted in blue, CFSE- cells are shown in red. The histograms in (M) and (O) show the proportion of CFSE+/- cells amongst viable CD4+FOXP3+CD127lo/- cells shown in (L) and (N). ‘Tnaive’ mix (L) and ‘Tmem’ mix (N) contained a mean of 14.73% and 4.35% CD4 cells expressing a Treg phenotype, respectively, out of which a mean of 17.6% (in the mix with ‘Tnaive’) and 23.6% (in the mix with ‘Tmem’) were CFSE neg, indicating the presence of contaminating Treg from either of the non-labelled T cell compartments (‘Tmem’ or ‘Tnaive’) or added ‘APC’.|
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