Enrichment of human Vγ9Vδ2 T lymphocytes by magnetic poly(divinylbenzene-co-glycidyl methacrylate) colloidal particles conjugated with specific antibody

RSC ADVANCES(2018)

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摘要
T cells play a significant role in protection against cancer. Purification of T cells is needed for insight when studying their anti-cancer functionality and their utilization in adoptive cell therapy. To improve the purification of T cells, in this work, a composite material based on magnetic nanoparticles was developed for purification of V9V2 T cells, the predominant subset of T lymphocytes in human peripheral blood. The epoxy-functionalized magnetic poly(divinylbenzene-co-glycidyl methacrylate) particles (mPDGs) were bio-conjugated with anti-human V2 antibody to provide specific recognition sites for T cell receptors of V9V2 T cells. Using fluorescence-activated cell sorting (FACS) analysis, separation of V9V2 T cells from peripheral blood mononuclear cells of healthy donors was confirmed with high purity [89.77% (range 87.00-91.80, n = 3)]. More interestingly, the immobilized particles did not affect the viability of purified cells as high cell viability was indicated (>90%). By combining the properties of magnetic nanoparticles with specific antibodies, these immobilized particles were shown to be used as a cell-friendly purification tool of V9V2 T lymphocytes without any limits for the further use of cells. The purified V9V2 T cells using the antibody-immobilized epoxy-functionalized mPDGs could be used directly without a detachment step for further cultivation and expansion. This highlights the advantages of this method in allowing the study of cell function and further investigation of such rare T cell populations in immunotherapy.
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