In Vitro Activation of NK Cells From Regional Lymph Nodes of Melanoma Patients With IL-2 and IL-15
2012
Аутори:
Vuletić, AnaJovanić, Irena
Nikolić, Srđan S
Tanić, Nikola
Konjević, Gordana
Остала ауторства
Eggermont, Alexander M.M.Тип документа:
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт:
Introduction: NK cells are subpopulation of innate immunity system and play
an important role in antitumor immune defense. Since regional lymph nodes
(LN) s represent the first barrier to malignant cell invasion, the aim of this study
was to investigate in melanoma (MM) patients, the ability of two cytokines,
IL-2, currently used for MM treatment, and IL-15 which is under intensive
clinical investigation and is essential for NK cell maturation, to modulate NK
cell antitumor activity of scarcely investigated NK cells from regional LNs.
Material and Method: Mononuclear cells were purified from 50 regional LNs
of MM patients and cultured for 72 h and 7 days in cell culture medium RPMI
1640 (CM) alone, CM with 200 IU/ml rhIL-2 and CM with 25 ng IL-15. NK cell
cytotoxicity was determined by standard 4h 51 Cr release assay, while perforine
(PRF) iRNA level was estimated by rt-PCR. Expression of several NK cell
receptors (Rc)s was analyzed after 7 day in vitro treatments on CD3− CD56+
NK cells by flow cytometry.
Results: Both cytokines induced significant in vitro enhancement of NK cell
cytotoxicity against K562 target tumor cell of tumor-infiltrated (LN+) and un-
infiltrated (LN−) LNs after 72 h of cultivation. Alongside with the induction of
NK cell cytotoxicity, the transcription of iRNA for cytotoxic mediator PRF was
significantly induced by each cytokine compared to control CM treatments.
After longer 7 day cultivation, the enhancement of NK cell cytotoxicity persisted
together with significant increase in expression of activating NKG2D and
cytotoxic CD16 Rcs, and CD69 early activation antigen on gated CD3− CD56+
NK cells from LN+ and LN−, induced by both cytokines. The analysis of
inhibitory KIR Rcs showed increase in CD158b KIR expression on NK cells
from LN+ and LN− by both cytokines. The expression of other investigated KIR
CD158a was induced by IL-15 only on NK cells from LN+. The comparison
of the cytokine induced NK cell cytotoxicity with respect to CM after 7 day in
vitro treatments, revealed the significantly lower increase in NK cell activity
obtained with IL-15 in LN+ compared to LN−, which is probably due to IL-15
induction of CD158a KIR that may act suppressive on NK cell cytotoxicity of
LN+.
Conclusion: IL-2 and IL-15 induce in vitro activation of NK cell from LNs
regardless the LN tumor infiltration probably by enhancing PRF transcription
and expression of NKG2D and CD16 activating Rcs on NK cells.
У:
- Eggermont AMM, editor. Proceedings Book: 22nd Biennial Congress of the European Association for Cancer Research; 2012 Jul 7-10; Barcelona, Spain. Oxford: Elsevier; 2012. p. S266. (European Journal of Cancer; Vol. 48; Suppl. 5).