BACKGROUND: In the past decade, human immunodeficiency virus (HIV) lymphocyte immunophenotyping has evolved significantly. New fluorochromes, new multicolor reagents, enhanced instruments, and the capacity to provide absolute cell counts using the single-platform technique have all contributed to the reliability of T-cell subset measurements. In this study, four gating protocols were evaluated to select the most robust method for T-cell subset enumeration. METHODS: Peripheral blood specimens from 21 HIV(+) and 20 HIV(-) individuals were monitored up to 96 h. Aliquots of specimens were stored at room temperature and analyzed at 6 (baseline), 48, 72, and 96 h. Aliquots were stained with CD45-fluorescein isothiocyanate (FITC)/CD3PC5/CD4RD1/CD8ECD. Data analysis was performed with all four gating protocols. RESULTS: Only with fresh blood did all protocols provide similar results. From samples that were 48 h old, the choice of gating strategy had a dramatic impact on immunophenotyping results. The largest deviations from baseline values occurred at 96 h and gating protocols that included dual light scatter gates provided the greatest shift of T-cell subset values over time. The gating protocols that were based exclusively on cell lineage-specific gates gave the most robust T-cell values up to 96 h. CONCLUSION: By selecting the appropriate gating protocol, the temporal integrity of specimens can be extended up to 4 days. Copyright 2002 Wiley-Liss, Inc.
BACKGROUND: In the past decade, human immunodeficiency virus (HIV) lymphocyte immunophenotyping has evolved significantly. New fluorochromes, new multicolor reagents, enhanced instruments, and the capacity to provide absolute cell counts using the single-platform technique have all contributed to the reliability of T-cell subset measurements. In this study, four gating protocols were evaluated to select the most robust method for T-cell subset enumeration. METHODS: Peripheral blood specimens from 21 HIV(+) and 20 HIV(-) individuals were monitored up to 96 h. Aliquots of specimens were stored at room temperature and analyzed at 6 (baseline), 48, 72, and 96 h. Aliquots were stained with CD45-fluorescein isothiocyanate (FITC)/CD3PC5/CD4RD1/CD8ECD. Data analysis was performed with all four gating protocols. RESULTS: Only with fresh blood did all protocols provide similar results. From samples that were 48 h old, the choice of gating strategy had a dramatic impact on immunophenotyping results. The largest deviations from baseline values occurred at 96 h and gating protocols that included dual light scatter gates provided the greatest shift of T-cell subset values over time. The gating protocols that were based exclusively on cell lineage-specific gates gave the most robust T-cell values up to 96 h. CONCLUSION: By selecting the appropriate gating protocol, the temporal integrity of specimens can be extended up to 4 days. Copyright 2002 Wiley-Liss, Inc.
Authors: George Janossy; Ilesh V Jani; Nicholas J Bradley; Arsene Bikoue; Tim Pitfield; Debbie K Glencross Journal: Clin Diagn Lab Immunol Date: 2002-09
Authors: Cécile Gouttefangeas; Cliburn Chan; Sebastian Attig; Tania T Køllgaard; Hans-Georg Rammensee; Stefan Stevanović; Dorothee Wernet; Per thor Straten; Marij J P Welters; Christian Ottensmeier; Sjoerd H van der Burg; Cedrik M Britten Journal: Cancer Immunol Immunother Date: 2015-02-18 Impact factor: 6.968
Authors: Michèle Bergeron; Géraldine Daneau; Tao Ding; Nadia E Sitoe; Larry E Westerman; Jocelijn Stokx; Ilesh V Jani; Lindi M Coetzee; Lesley Scott; Anja De Weggheleire; Luc Boel; Wendy S Stevens; Deborah K Glencross; Trevor F Peter Journal: PLoS One Date: 2012-08-09 Impact factor: 3.240