CMV-specific T cell isolation from G-CSF mobilized peripheral blood: depletion of myeloid progenitors eliminates non-specific binding of MHC-multimers

Table 1 Phenotypic characterization and CMV-specific CTL isolation of unmanipulated PBMCs and the non-adherent fraction

	Unmanipulated PBMCs	Non-adherent fraction	p
	Leukocyte subpopulations
CD14+ cells	47.8% (36.0 - 55.0)	2.1% (1.3 - 6.1)	0.005
T lymphocytes (CD3+)	24.0% (18.0 - 37.1)	69.9% (31.9 - 79.2)	0.007
CTLs (CD3 + CD8+)	17.7% (6.8 - 25.1)	35.2% (23.8 - 45.2)	0.009
Helper T cells (CD3 + CD4+)	12.5% (11.4 - 19.4)	19.3% (16.6 - 29.6)	0.208
Viability (7-AAD-)	98.0% (95.6 - 98.5)	96.0% (93.4 - 97.8)	0.066
	PM staining: specific and non-specific binding
CMV-specific CTLs (CD3 + CD8 + PM+)	0.14% (0.06 - 0.62)	0.65% (0.24 - 1.51)	0.003
Non-specific PM binding (CD8-PM+)	0.56% (0.41 - 0.86)	0.16% (0.12 - 0.37)	0.003
	CMV-specific CTL isolation
Purity	20.8% (6.9 - 61.7)	76.0% (32.7 - 83.7)	0.043
Yield	38.6% (32.9 - 44.8)	42.1% (23.4 - 84.1)	0.893

Cell subsets, CMV-specific CTLs and non-specific PM staining were analyzed in the unmanipulated PBMC sample and in the non-adherent cells (n = 11). CMV-specific CTLs were isolated from unmanipulated PBMCs and non-adherent fraction, and purity and yield of the obtained cellular product were determined (n = 5). Comparison was done with the Wilcoxon signed-rank test and significance level was fixed to p < 0.05.

ISSN: 1479-5876