Peripheral blood lymphocytes (PBLs) from HLA-A2 positive donors were collected as previously described21 (link). Briefly, leukapheresis (Spectra Optia®, TerumoBCT) was performed on consenting healthy donors (Ethical license B.U.N. 143201420996) in accordance with the relevant guidelines and regulations. The leukapheresis product was processed via an elutriation procedure to collect monocytes (Elutra®, TerumoBCT). The remaining PBLs were counted and cryopreserved in 5% DMSO-containing cryopreservation medium (CryoStor® CS5, Biolife Solutions) till further use.
To study the contribution of NK cells to the observed effects, PBLs were depleted from CD19+ cells using anti-CD19 magnetic beads to enhance purity of NK, CD4+ and CD8+ T cells within the PBLs. Subsequently, CD4+ and CD8+ T cells were depleted with respectively anti-CD4 and anti-CD8 magnetic beads. To study the contribution of T cells to the observed effects, PBLs were selected for CD4+ and CD8+ T cells using anti-CD4 and anti-CD8 magnetic beads. Depletions and selections were performed according to the manufacturer’s instructions.
To study the contribution of NK cells to the observed effects, PBLs were depleted from CD19+ cells using anti-CD19 magnetic beads to enhance purity of NK, CD4+ and CD8+ T cells within the PBLs. Subsequently, CD4+ and CD8+ T cells were depleted with respectively anti-CD4 and anti-CD8 magnetic beads. To study the contribution of T cells to the observed effects, PBLs were selected for CD4+ and CD8+ T cells using anti-CD4 and anti-CD8 magnetic beads. Depletions and selections were performed according to the manufacturer’s instructions.
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