Interleukin-2 activated NK cells do not use the CD95L- and TRAIL-pathways in the rapid induction of apoptosis of rat colon carcinoma CC531s cells

Authors: Velthuis J.H.L.¹; de Bont H.J.G.M.¹; Medema J-P.²; Kuppen P.J.K.³; Mulder G.J.¹; Nagelkerke J.F.¹

Source: Immunobiology, Volume 207, Number 2, March 2003 , pp. 115-127(13)

Publisher: Urban & Fischer

Abstract:

Natural Killer (NK) cells can induce apoptosis in target cells in at least four ways: by secretion of granzyme B/perforin (GrB/P) and via the CD95L, TRAIL and TNF- pathways. In this study we examined the pathways used by interleukin-2 activated rat NK (A-NK) cells to induce apoptosis in the rat colon carcinoma cell line CC531s. Co-incubation of A-NK cells with CC531s cells for three hours resulted in 70% apoptosis in the latter. Addition of the GrB/P pathway-inhibitor concanamycin A reduced the number of apoptotic cells to 54%. Blockade of the CD95L, TRAIL and TNF- pathways by specific antibodies hardly had an additional effect. However, co-incubation with transfected MEC cells that expressed CD95L or 2PK3-cells that expressed TRAIL did induce apoptosis in CC531s cells. Furthermore the A-NK cells contained CD95L and TRAIL. However, comparison of non- and permeabilized cells revealed that the majority of TRAIL was present in the cytosol of A-NK cells and was not available for induction of apoptosis. The presence of elevated levels of bcl-2 in CC531 cells reduced the sensitivity towards induction of apoptosis both by A-NK cells as well as the CD95L and TRAIL expressing cell lines. Using the caspase-inhibitors ac-IEPD-CHO, ac-DEVD-CHO and zVAD-fmk, it was shown that inhibition of the effector caspase-3 prevented A-NK cell induced apoptosis in CC531-bcl-2 cells, but not in CC531s cells. In conclusion, A-NK cells kill by secretion of GrB/P and not by the CD95L, TRAIL or TNF pathways albeit both CD95L and TRAIL are produced by the A-NK cells.

Language: English

Document Type: Original article

DOI: http://dx.doi.org/10.1078/0171-2985-00226

Affiliations: 1: Division of Toxicology, Leiden/Amsterdam Center for Drug Research, Leiden University, The Netherlands 2: Department of Immunohematology and Bloodbank, LUMC, Leiden, The Netherlands 3: Department of Surgery, LUMC, Leiden, The Netherlands

Publication date: 2003-03-01

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• By this: publisher
• In this Subject: Biotechnology ,  Microbiology ,  Pharmacology
• By this author: Velthuis J.H.L. ;  de Bont H.J.G.M. ;  Medema J-P. ;  Kuppen P.J.K. ;  Mulder G.J. ;  Nagelkerke J.F.

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