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  • Title: Natural killer cell numbers and activity in mobilized peripheral blood stem cell grafts: conditions for in vitro expansion.
    Author: Silva MR, Parreira A, Ascensão JL.
    Journal: Exp Hematol; 1995 Dec; 23(14):1676-81. PubMed ID: 8542964.
    Abstract:
    Peripheral blood stem cells (PBSC) are increasingly being used as an alternative to autologous bone marrow (BM) for hematologic rescue after high-dose chemoradiotherapy in the treatment of hematologic and nonhematologic malignancies. Mobilization procedures such as chemotherapy and/or hematopoietic growth factor administration are employed to allow for the graft enrichment in hematopoietic stem cells and progenitors and to accelerate trilineage recovery after transplant. The influence of these mobilization procedures on the lymphoid populations in the graft and on immunologic recovery after transplant remains to be determined. We studied six consecutive patients undergoing PBSC high-volume collections after cyclophosphamide (Cyc) and granulocyte colony-stimulating factor (G-CSF) administration and observed that NK cell numbers (phenotypically defined as CD3-CD56+ by flow cytometry) and activity (evaluated by a 51Cr release assay) fully recovered after 4-5 weeks; high numbers of functionally active NK cells (42.1-212.1 x 10(6)/kg b.w.) were present in the grafts, and their percentage and cytotoxic activity rose from the beginning to the end of the harvesting procedure in most cases. CD3-CD56+ and CD34+ cell numbers peaked at the same time point during harvesting, which differed from one patient to another. T (CD3+) cells were always present during harvest, and CD4 and CD8 numbers showed interdonor variability. When we cultured leukapheresed PBSC in the presence of interleukin-2 (IL-2) (10-1000 U/mL) for 6-8 days, we were able to expand the NK population three- to 5.4-fold; 100 U/mL appears to be the best concentration to generate high numbers of cytotoxic NK cells. Pilot studies also suggest that this short exposure to IL-2 does not affect the CD34+ cells. We conclude that PBSC grafts mobilized by combined Cyc and G-CSF and harvested through high-volume leukapheresis contain high numbers of cytotoxic NK cells that can be expanded in vitro by exposure to IL-2. In the setting of PBSC transplant, ex vivo immunomodulation aimed at increasing the NK cell numbers and activity is feasible and may prove to be useful in inducing a graft-vs.-tumor effect, thereby decreasing the relapse rate after transplant.
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