Use of interleukin-15 for preparation of adherent NK cells from human peripheral blood: comparison with interleukin-2.
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 51Chromium 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 (IFNgamma) 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.
Sun
Rui
R
School of Life Sciences, University of Science and Technology of China, Anhui 230027, Hefei, China.
Fan
Jing
J
Wei
Haiming
H
Zhang
Cai
C
Tian
Zhigang
Z
D015415:Biomarkers; D002448:Cell Adhesion / Q000502:physiology; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D019409:Interleukin-15 / Q000378:metabolism*; D007376:Interleukin-2 / Q000378:metabolism*; D007694:Killer Cells, Natural / Q000378:metabolism*; D007700:Kinetics
D015415:Biomarkers; D016207:Cytokines; D019409:Interleukin-15; D007376:Interleukin-2