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Clinical and Vaccine Immunology, June 2007, p. 775-781, Vol. 14, No. 6
1071-412X/07/$08.00+0 doi:10.1128/CVI.00043-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.
Evaluation of a Simplified Dual-Platform Flow Cytometric Method for Measurement of Lymphocyte Subsets and T-Cell Maturation Phenotypes in the Population of Nouna, Burkina Faso
T. Böhler,1*
M. von Au,1
N. Klose,1
K. Müller,1
B. Coulibaly,3
F. Nauwelaers,4
H. P. Spengler,4
G. Kynast-Wolf,2 and
H.-G. Kräusslich1
Department of Virology,1 Department of Tropical Medicine and Public Health, Institute of Hygiene, University of Heidelberg, Heidelberg, Germany,2 Nouna Health Research Centre, Nouna, Burkina Faso,3 BD Biosciences, Erembodegem, Belgium4
Received 21 January 2007/ Returned for modification 1 March 2007/ Accepted 22 March 2007
In the context of a larger clinical study in Nouna, Burkina Faso, we evaluated a simplified dual-platform (DP) flow cytometric (FCM) method that allows the determination of major lymphocyte subsets in a single test tube. We compared the phenotyping of lymphocytes with DP FCM and simultaneous measurements with standard single-platform (SP) FCM for samples from 177 individuals. Analysis of the comparative measurements revealed that DP FCM systematically underestimates the proportion of NK cells, overestimates the percentage of CD3+ CD8+ lymphocytes, and yields proportions of B cells and CD4+ T cells comparable with the results from SP FCM. Bland-Altman analysis showed a low bias between both methods and an acceptable precision for percent values of CD4+ T cells (bias ± precision, –1% ± 6%) and CD8+ T cells (–3% ± 6%). The absolute cell numbers of all lymphocyte subpopulations, however, were systematically biased towards lower values being obtained by DP FCM. Reference values for the distribution of T-cell maturation phenotypes in 177 healthy adults were calculated using DP FCM. The mean ± standard deviation (SD) CD4+-to-CD8+ T-cell ratio was 1.61 ± 0.61, the mean percentage ± SD of CD4+ T cells was 42% ± 7%, and that of CD8+ T cells 29% ± 7%. Among CD4+ lymphocytes, 28% ± 7% were classified as central memory (CD45RAlow CCR7+), 22% ± 10% as naïve (CD45RAhigh CCR7+), 45% ± 12% as effector memory (CD45RAlow CCR7–); and 5% ± 3% as terminally differentiated effector memory expressing CD45RA (CD45RAhigh CCR7–). Among CD8bright lymphocytes, 3% ± 2% had a central memory phenotype, 27% ± 13% were naïve, 37% ± 13% had an effector memory phenotype, and 34% ± 12% were terminally differentiated effector memory cells expressing CD45RA.
* Corresponding author. Mailing address: c/o Department of Virology, University of Heidelberg, Im Neuenheimer Feld 324, D-69120 Heidelberg, Germany. Phone: 49-6221 565002. Fax: 49-6221 565003. E-mail: thomas.boehler{at}med.uni-heidelberg.de
Published ahead of print on 18 April 2007.
Clinical and Vaccine Immunology, June 2007, p. 775-781, Vol. 14, No. 6
1071-412X/07/$08.00+0 doi:10.1128/CVI.00043-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.
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