Functional role of granzymes in epidermal γδ T cell-mediated killing of tumor targets

M. Mohamadzadeh, M. J. McGuire, D. J. Smith, P. R. Bergstresser, A. Takashima

Research output: Contribution to journalArticle

Abstract

Granzymes, a family of serine proteases contained in cytoplasmic granules, play a critical role in killing tumor targets, by triggering the rapid breakdown of DNA and subsequent apoptosis. Dendritic epidermal T cells (DETC), skin-specific members of the tissue-type γδ T cell family in mice, have been shown to kill selected tumor cell lines. Our purpose was, therefore, to elucidate the functional role of granzymes in DETC-mediated killing. The short-term DETC line (C25) established from CBA mice: a) lysed YAC-1 lymphoma targets effectively (51Cr release assay), b) contained significant BLT esterase activity (colorimetric enzyme analysis), c) expressed constitutively mRNA for granzymes A and B (Northern blotting), and d) produced relatively large amounts of granzyme A (immunoblotting). mRNA expression for granzyme B was confirmed by RT-PCR analysis of Thy-l+ epidermal cells (i.e., DETC) that were freshly isolated from CBA mice. C25 DETC also induced apoptosis of the YAC-1 targets as measured by their capacity to cause breakdown of 3H-thymidine-labeled DNA. Importantly, this ability was blocked completely by preincubation of DETC with the granzyme inhibitor, dichloroisocoumarin (DCI), but not with the cysteine protease inhibitor, E-64. Thus, we conclude that granzymes play an essential role in DETC-mediated apoptosis of tumor targets.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - 1996

Fingerprint

Granzymes
T-cells
Tumors
T-lymphocytes
T-Lymphocytes
neoplasms
Neoplasms
Inbred CBA Mouse
apoptosis
Apoptosis
mice
cell lines
cytoplasmic granules
Cytoplasmic Granules
Cysteine Proteinase Inhibitors
cysteine proteinase inhibitors
Messenger RNA
Langerhans Cells
DNA
Serine Proteases

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Mohamadzadeh, M., McGuire, M. J., Smith, D. J., Bergstresser, P. R., & Takashima, A. (1996). Functional role of granzymes in epidermal γδ T cell-mediated killing of tumor targets. FASEB Journal, 10(6).

Functional role of granzymes in epidermal γδ T cell-mediated killing of tumor targets. / Mohamadzadeh, M.; McGuire, M. J.; Smith, D. J.; Bergstresser, P. R.; Takashima, A.

In: FASEB Journal, Vol. 10, No. 6, 1996.

Research output: Contribution to journalArticle

Mohamadzadeh, M, McGuire, MJ, Smith, DJ, Bergstresser, PR & Takashima, A 1996, 'Functional role of granzymes in epidermal γδ T cell-mediated killing of tumor targets', FASEB Journal, vol. 10, no. 6.
Mohamadzadeh M, McGuire MJ, Smith DJ, Bergstresser PR, Takashima A. Functional role of granzymes in epidermal γδ T cell-mediated killing of tumor targets. FASEB Journal. 1996;10(6).
Mohamadzadeh, M. ; McGuire, M. J. ; Smith, D. J. ; Bergstresser, P. R. ; Takashima, A. / Functional role of granzymes in epidermal γδ T cell-mediated killing of tumor targets. In: FASEB Journal. 1996 ; Vol. 10, No. 6.
@article{4565f7cac94a4c62857d7c821578a27f,
title = "Functional role of granzymes in epidermal γδ T cell-mediated killing of tumor targets",
abstract = "Granzymes, a family of serine proteases contained in cytoplasmic granules, play a critical role in killing tumor targets, by triggering the rapid breakdown of DNA and subsequent apoptosis. Dendritic epidermal T cells (DETC), skin-specific members of the tissue-type γδ T cell family in mice, have been shown to kill selected tumor cell lines. Our purpose was, therefore, to elucidate the functional role of granzymes in DETC-mediated killing. The short-term DETC line (C25) established from CBA mice: a) lysed YAC-1 lymphoma targets effectively (51Cr release assay), b) contained significant BLT esterase activity (colorimetric enzyme analysis), c) expressed constitutively mRNA for granzymes A and B (Northern blotting), and d) produced relatively large amounts of granzyme A (immunoblotting). mRNA expression for granzyme B was confirmed by RT-PCR analysis of Thy-l+ epidermal cells (i.e., DETC) that were freshly isolated from CBA mice. C25 DETC also induced apoptosis of the YAC-1 targets as measured by their capacity to cause breakdown of 3H-thymidine-labeled DNA. Importantly, this ability was blocked completely by preincubation of DETC with the granzyme inhibitor, dichloroisocoumarin (DCI), but not with the cysteine protease inhibitor, E-64. Thus, we conclude that granzymes play an essential role in DETC-mediated apoptosis of tumor targets.",
author = "M. Mohamadzadeh and McGuire, {M. J.} and Smith, {D. J.} and Bergstresser, {P. R.} and A. Takashima",
year = "1996",
language = "English (US)",
volume = "10",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "6",

}

TY - JOUR

T1 - Functional role of granzymes in epidermal γδ T cell-mediated killing of tumor targets

AU - Mohamadzadeh, M.

AU - McGuire, M. J.

AU - Smith, D. J.

AU - Bergstresser, P. R.

AU - Takashima, A.

PY - 1996

Y1 - 1996

N2 - Granzymes, a family of serine proteases contained in cytoplasmic granules, play a critical role in killing tumor targets, by triggering the rapid breakdown of DNA and subsequent apoptosis. Dendritic epidermal T cells (DETC), skin-specific members of the tissue-type γδ T cell family in mice, have been shown to kill selected tumor cell lines. Our purpose was, therefore, to elucidate the functional role of granzymes in DETC-mediated killing. The short-term DETC line (C25) established from CBA mice: a) lysed YAC-1 lymphoma targets effectively (51Cr release assay), b) contained significant BLT esterase activity (colorimetric enzyme analysis), c) expressed constitutively mRNA for granzymes A and B (Northern blotting), and d) produced relatively large amounts of granzyme A (immunoblotting). mRNA expression for granzyme B was confirmed by RT-PCR analysis of Thy-l+ epidermal cells (i.e., DETC) that were freshly isolated from CBA mice. C25 DETC also induced apoptosis of the YAC-1 targets as measured by their capacity to cause breakdown of 3H-thymidine-labeled DNA. Importantly, this ability was blocked completely by preincubation of DETC with the granzyme inhibitor, dichloroisocoumarin (DCI), but not with the cysteine protease inhibitor, E-64. Thus, we conclude that granzymes play an essential role in DETC-mediated apoptosis of tumor targets.

AB - Granzymes, a family of serine proteases contained in cytoplasmic granules, play a critical role in killing tumor targets, by triggering the rapid breakdown of DNA and subsequent apoptosis. Dendritic epidermal T cells (DETC), skin-specific members of the tissue-type γδ T cell family in mice, have been shown to kill selected tumor cell lines. Our purpose was, therefore, to elucidate the functional role of granzymes in DETC-mediated killing. The short-term DETC line (C25) established from CBA mice: a) lysed YAC-1 lymphoma targets effectively (51Cr release assay), b) contained significant BLT esterase activity (colorimetric enzyme analysis), c) expressed constitutively mRNA for granzymes A and B (Northern blotting), and d) produced relatively large amounts of granzyme A (immunoblotting). mRNA expression for granzyme B was confirmed by RT-PCR analysis of Thy-l+ epidermal cells (i.e., DETC) that were freshly isolated from CBA mice. C25 DETC also induced apoptosis of the YAC-1 targets as measured by their capacity to cause breakdown of 3H-thymidine-labeled DNA. Importantly, this ability was blocked completely by preincubation of DETC with the granzyme inhibitor, dichloroisocoumarin (DCI), but not with the cysteine protease inhibitor, E-64. Thus, we conclude that granzymes play an essential role in DETC-mediated apoptosis of tumor targets.

UR - http://www.scopus.com/inward/record.url?scp=33749126794&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33749126794&partnerID=8YFLogxK

M3 - Article

VL - 10

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 6

ER -