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Use of interleukin-15 for preparation of adherent NK cells from human peripheral blood: comparison with interleukin-2

Journal of Immunological Methods, ISSN: 0022-1759, Vol: 279, Issue: 1, Page: 79-90
2003
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Article Description

To search the possibility of utilizing interleukin-15 (IL-15) in preparation of adherent human natural killer (A-NK) cells, recombinant human IL-15 (rhIL-15) or rhIL-2 (500 u/ml of each cytokine) were added to purified human NK cell culture in 24-well plastic plate. The cytokine-induced adherent ratio was calculated by percentage of A-NK cell in whole NK cells. The cytotoxicity of NK cells (NA- or A-NK cells) was examined by 4-h 51 Chromium release assay, the surface markers of NK cells were checked by flow cytometry, and the cytokines were analyzed by reverse transcript (RT)-PCR and ELISA method. RhIL-15-induced adherence of human NK cells into plastic was higher than IL-2 when harvesting the A-NK cells at each hour point from hr 1 to hr 12. IL-15- and IL-2-induced adherent ratio peaked to 36.67% and 27.73% at hr 1, and the IL-15-induced adherent ratio was around two folds higher than IL-2-induced group at hrs 2, 3, 4, 5, 6, 7, and 8. The IL-15 group expanded more rapidly than IL-2 during 2 weeks' culture. IL-15- and IL-2-A-NK cells exerted similar levels of higher cytotoxic potentials. A-NK cells were characterized with phenotypes of CD3 − CD16 + CD56 + (more than 93%) in the presence of IL-2 or IL-15 stimulation. CD54, an intracellular adhesion molecule (ICAM), was also continuously expressed in A-NK cells (more than 85%) induced by each cytokine. Interestingly, IL-15 stimulated relatively low level of expression of CD18, a beta2 integrin molecule related to lymphocyte apoptosis in A-NK cells (11.45%), whereas IL-2 exerted a strong effect on CD18 expression (87.54%). IL-11b was only expressed at A-NK cell induced by IL-2 (49.56%), IL-15 did not exert any stimulating effect on CD11b expression. All A-NK cells expressed high levels of interferon gamma (IFNγ) after stimulation with IL-2 or IL-15. In contrast to IL-2, IL-15 did not stimulate gene expressions of type 2 cytokines (e.g. IL-4, IL-6, IL-10 and IL-13) in A-NK cells. The results indicate that rhIL15 is possibly a stronger stimulator for A-NK cell preparation by improving adherence and proliferation through inhibiting apoptosis by down-regulating the expression of CD18 and type 2 cytokines.

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