Project description:NK cells were isolated from the perpheral blood of healthy donors (n=6) and expanded ex vivo in the presence of feeder cells and IL-2. RNA from fresh and expanded cells was isolated, sequenced, and gene expression of the cell populations were compared.
Project description:NK cells are cytotoxic lymphocytes that play an important role in the innate immune response. The immune response of NK cells was shown under activated conditions. In order to identify such patterns, we performed RNA sequencing to confirm the gene expression profiles in NK cells expanded ex vivo compared to that of resting NK cells.
Project description:Ex vivo activation and expansion of natural killer (NK) cells is a strategy to produce sufficient numbers of these effector cells for adoptive immunotherapy. Therefore we aimed at the development of an automated, clinical scale NK cell expansion process and compared NK cells after manual and automated expansion.We found only a small subset of 124 genes that significantly varied between both sample groups. These genes were associated with movement of leukocytes were identified including a group of genes for NK cell movement (CMKLR1, CX3CR1, S1PR5, GNLY and CXCR1) which were slightly higher expressed after automated compared to manual expansion. Nevertheless, the expansion profiles of NK cells after automated or manual expansion were rather similar in comparison to the remarkable difference between primary and expanded NK cells in general represented by the vast majority of regulated genes between the analyzed sample groups. Pathway analysis revealed that most of regulated genes upon expansion were associated with cell cycle regulation, DNA replication, DNA recombination and DNA repair, regulation of apoptosis and cell survival as well as cytokine signaling. Furthermore, the expression of many NK cell relevant markers was changed upon expansion with the strongest effects on up regulation of TRAIL and FASL, inhibitory TIGIT and chemokine receptors CCR2, CCR5 and CXCR6. In addition, granzyme M was down regulated but other important effector molecules like TNFa, perforin and granzymes A, B and K were up regulated. In summary we could show that manual and automatically expanded NK cells show a similar expansion profile while the differ significantly from primary NK cells. Up regulation of many NK cell relevant markers indicate for an enhanced NK cell functionaility after ex vivo activation and expansion. 24 samples were analyzed in total. 6 biological replicates represented by cells from different donors. 4 different samples per donor: freshly isolated NK cells (d0) and NK cells expanded for 14 days under 3 different conditions: in co-culture with EBV-LCL feeder cells and 500 U/mL IL2 either cultivated manually in T75 flasks (T) or automatically using the CliniMACS Prodigy system (P); NK cells cultured with 500 U/ml IL2 without feeder cells (I)
Project description:RNA-seq analysis was performed to compare differentially expressed genes in freshly isolated and ex-vivo cultured human cord blood CD34+ cells. Mitochondrion related genes are upregulated in CD34+ hematopoietic stem and progenitor cells upon ex vivo culture. In vivo transplantation experiments demonstrate that stemness of CD34+ cells is significantly decreased due to oxidative stress induced by ex vivo culture.
Project description:we performed RNA sequencing to employ an unbiased approach to identify the altered gene expression enhancing the function of ex vivo expanded pNK cells for 14 days, and transcriptionally compared day 0 and day 14 pNK cells.
Project description:We performed RNA sequencing to employ an unbiased approach to identify the altered gene expression enhancing the function of ex vivo expanded pNK cells for 14 days, and transcriptionally compared day 0 and day 14 pNK cells.
Project description:A study of immunotherapy with expanded CB-NK cells in combination with cetuximab, to evaluate activity against minimal residual disease in patients with colon cancer that have completed adjuvant treatment but are positive for ctDNA