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|Title:||The effects of anti-CD3/CD28 coated beads and IL-2 on expanded T cell for immunotherapy|
|Keywords:||Biochemistry, Genetics and Molecular Biology;Medicine|
|Citation:||Advances in Clinical and Experimental Medicine. Vol.25, No.5 (2016), 821-828|
|Abstract:||© by Wroclaw Medical University. Background: The activation of peripheral blood mononucleated cells (PBMCs) with anti-CD3/CD28-coated magnetic beads promotes intrinsic resistance to HIV as well as cell expansion. Objectives: The aim of this study was to define an optimal cell isolation protocol for the expansion of PBMCs using anti-CD3/CD28-coated bead stimulation, with the ultimate goal of using these cells for adoptive therapy. Material and Methods: PBMCs were isolated from healthy donor blood samples. To determine the effect of varying the bead-to-cell ratios on the expansion rate and phenotypic characterization of the expanded cells, one million PBMCs were stimulated by anti-CD3/CD28-coated beads at bead-to-cell ratios of 0.1:1, 0.5:1 and 1.0:1 in the presence of 100 U/mL exogenous IL-2; also, one million PBMCs were stimulated by anti-CD3/CD28-coated beads at a bead-to-cell ratio of 0.5:1 in the presence of varying concentrations of IL-2 (20, 100 and 1000 U/mL). Cell expansion was carried out for three weeks. The cell numbers, cell viability and phenotypic characterization were determined by trypan blue exclusion and flow cytometry. Results: The initial experiments showed no difference in the expansion rate from cells grown with the three different bead-to-cell ratios. PBMCs can be expanded up to 1000-fold at a 0.5:1 bead-to-cell ratio after three weeks of cell expansion with a high viability (90%). Furthermore, in the presence of 100 U/mL IL-2, the percentages of CD3-CD16+CD56+ cells showed marked increases. Conclusions: The results demonstrate that PBMCs were stimulated with anti-CD3/CD28-coated beads. This method may provide an alternative for driving T cell expansion, which may be very useful in adoptive immunotherapy.|
|Appears in Collections:||Scopus 2016-2017|
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