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Soluble factors released by activated cytotoxic T lymphocytes interfere with death receptor pathways in neuroblastoma

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Abstract

Neuroblastoma (NB) is often described as an unfavorable target for both HLA-restricted and death receptor-mediated elimination by cytotoxic T lymphocytes (CTLs) due to low or absent HLA class I and caspase-8 expression. We investigated the effects of soluble factors released by CTLs activated by TCR triggering (named as activated supernatant; AS) on the levels and composition of cell surface molecules involved in HLA-restricted and HLA-independent NB cell recognition (surface immune phenotype). Using a panel of long-term propagated NB cell lines and freshly isolated primary human NB cells, we analyzed surface expression of the (1) cognate receptors for TNFα, Fas and TRAIL; (2) HLA class I and II heterodimers; (3) adhesion molecules; (4) the intracellular expression and activation of caspase-8, as well as (5) the susceptibility of NB cells to death receptor-mediated killing prior to and after exposure to AS. The exposure of NB cells to soluble factors released by activated CTLs skewed the surface immune phenotype of both long term cultured and primary NB cells, induced the expression and activation of caspase-8 and increased the susceptibility of tumor cells to lysis by TRAIL and Fas-agonistic antibody. Blocking experiments identified IFNγ and TNFα as main factors responsible for modulating the surface antigens of NB cells by AS. Our data suggest that recruitment of CTLs activated on third party targets into the vicinity of the NB tumor mass, may override the “silent” immune phenotype of NB cells via the action of soluble factors.

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Abbreviations

AS:

Activated supernatant

APC:

Allophycocyanin

β2m:

β2-Microgloblin

CS:

Control supernatant

CTL:

Cytotoxic T lymphocyte

EBV:

Epstein Barr virus

FITC:

Fluorescein isothiocyanate

FLICA:

Fluorochrome inhibitor of caspases

HC:

Heavy chain

HLA:

Human leukocyte antigen

IFN:

Interferon

MICA:

MHC class I chain-related A

NB:

Neuroblastoma

RPE:

R-phycoerythrin

TCR:

T-cell receptor

TNF-R:

TNFα-receptor

TRAIL:

Tumor necrosis factor-related apoptosis-inducing ligand

v/v:

Volume-to-volume

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Authors and Affiliations

  1. Department of Oncology-Pathology, Cancer Centrum Karolinska, Karolinska Institutet, KS-ringen R8:01, 17176, Stockholm, Sweden

    Anna De Geer, Lena-Maria Carlson & Jelena Levitskaya

  2. Department of Woman and Child Health, Childhood Cancer Research Unit, Astrid Lindgren Children’s Hospital, Q6:05, 17176, Stockholm, Sweden

    Per Kogner

  3. W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD, 21205, USA

    Jelena Levitskaya

Authors
  1. Anna De Geer
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  2. Lena-Maria Carlson
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  3. Per Kogner
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  4. Jelena Levitskaya
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Corresponding author

Correspondence to Jelena Levitskaya.

Additional information

This work was supported by grants from the Swedish Children’s Cancer Foundation, the Swedish Cancer Society, the Cancer Society of Stockholm, the King Gustav the Vth Jubilee Fund, Karolinska Institutet and the Swedish Research Council.

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De Geer, A., Carlson, LM., Kogner, P. et al. Soluble factors released by activated cytotoxic T lymphocytes interfere with death receptor pathways in neuroblastoma. Cancer Immunol Immunother 57, 731–743 (2008). https://doi.org/10.1007/s00262-007-0412-2

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