Blood collection tubes impact expression of activated CD4+ and CD8+ T cells in human whole blood assay

Background: T-lymphocyte subsets CD4 and CD8 play important role in host immune responses. However, little attention has been given to the impact of time lapse and the various anticoagulant blood collection tubes on the expression frequency and activation status of CD4+ and CD8+ T cells. To this end, we explore the impact of time (t<1 h and t=4 h) and collection tubes (EDTA and heparin) on the expression frequency and activation status of CD4+ and CD8+ T cells among healthy Ghanaian individuals.Methods: A cohort of healthy individuals (n=9) is recruited, and blood samples obtained in Ghana for the frequency of CD4+and CD8+ T cells at various time points (<1 h and 4 h). The proportions of activation of these immune markers were profiled using immunophenotyping.Results: Significant statistical differences in the activation frequency of CD69 expressing CD4+T cells (t < 1 h and t=4 h; p=0.02) and CD69 expressing CD8+ T cells from EDTA tubes at times (t < 1 h and t=4 h; p=0.05) was observed. No significant difference were observed with CD69 expressing cells in Heparin tubes. Notably, CD8+ T cell activation frequency was observed to be consistently higher than that of CD4+ T cell at the various study time points and in the collection tubes used. No marked alterations were observed witth the proportion of CD4+ and CD8+ T cells in the samples collected at the time points; <1 h and at 4 h.Conclusion: The study shows that activation of CD4+ and CD8+ T cells in EDTA tubes differed significantly between both time points (t <1 h and t=4 h) but not in the heparin collection tubes. Therefore, it is important to take into account the elapsed time and the type of blood collection tubes when performing phenotypic characterization of activated immune markers.