Granulysin, a newly described cytolytic protein, may contribute to the tumorolytic effect of nkt cells

Jennifer L. Gansert, Maria T. Ochoa, Alan M. Krensky, Steven A. Porcelli, Robert L. Modlin

Research output: Contribution to journalArticle

Abstract

Human NKT cells recognize the glycolipid antigen, a-galactosylceramide (aGalCer), presented in the context of the non-classical antigen-presenting molecule, CD Id. However, NKT cells exposed to aGalCer can lyse a number of CD Id-negative tumor-derived ;ell lines. Lysis of these tumor targets in the absence of CDld suggests that aGalCer-activuted NKT cells may release soluble mediators. One possible mediator is granulysin, a newly described component of cytolytic granules present in activated cytotoxic T cells and NK cells. Granulysin is a member of the saposin-like protein family that has direct antimicrobial activity and that can activate caspase 9 to induce apoptosis in nucleated cells. In the present study, immunofluorescent staining demonstrated a granular pattern of granulysin expression in NKT cells. Purified granulysin protein was tested against a number of ta'get cells and correlated with results of cytotoxicity assays. The CDld-positive Molt-3 T cell acute lymphoblastic leukemia (ALL) cell line was specifically lysed by an NKT cell clone derived from a normal donor at the effector to target ratio of 5:1 (40% specific lysis) and was sensitive to lysis by granulysin (30% increase in propidium iodide-staining cells;. In contrast, a recently derived CD Id-negative T cell leukemia cell line was resistant to both lysis by NKT cells (3% specific lysis) and to lysis by granulysin (10% lysis). This cell line was established from a patient with chemo-refractory disease and contairs a chromosomal rearrangement at Ip36.1, the location of the caspase 9 gene. Caspase 9 is required for apoptosis induced by chemotherapeutic agents, as well as by granulysin. Loss of caspase 9 activity in this cell line is being investigated. Finally, primary ALL cells from an untreated patient were sensitive to granulysin (30% lysis). Overall, these studies suggest that the tumorolytic effect of aGalCer-activated NKT cells may be mediated through the release of soluble cytotoxic mediators, such as granulysin, and that the optima) timing of immunotherapy using the NKT cell/aGalCer system is early in disease, before resistance emerges.

Original languageEnglish (US)
JournalBlood
Volume96
Issue number11 PART I
StatePublished - 2000

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Galactosylceramides
Natural Killer T-Cells
Caspase 9
T-cells
Cells
Proteins
Tumors
Clone cells
Cell Line
Saposins
Apoptosis
Antigens
Propidium
Glycolipids
Cytotoxicity
Refractory materials
Staining and Labeling
Assays
T-Cell Leukemia
Precursor T-Cell Lymphoblastic Leukemia-Lymphoma

ASJC Scopus subject areas

  • Hematology

Cite this

Gansert, J. L., Ochoa, M. T., Krensky, A. M., Porcelli, S. A., & Modlin, R. L. (2000). Granulysin, a newly described cytolytic protein, may contribute to the tumorolytic effect of nkt cells. Blood, 96(11 PART I).

Granulysin, a newly described cytolytic protein, may contribute to the tumorolytic effect of nkt cells. / Gansert, Jennifer L.; Ochoa, Maria T.; Krensky, Alan M.; Porcelli, Steven A.; Modlin, Robert L.

In: Blood, Vol. 96, No. 11 PART I, 2000.

Research output: Contribution to journalArticle

Gansert, JL, Ochoa, MT, Krensky, AM, Porcelli, SA & Modlin, RL 2000, 'Granulysin, a newly described cytolytic protein, may contribute to the tumorolytic effect of nkt cells', Blood, vol. 96, no. 11 PART I.
Gansert, Jennifer L. ; Ochoa, Maria T. ; Krensky, Alan M. ; Porcelli, Steven A. ; Modlin, Robert L. / Granulysin, a newly described cytolytic protein, may contribute to the tumorolytic effect of nkt cells. In: Blood. 2000 ; Vol. 96, No. 11 PART I.
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abstract = "Human NKT cells recognize the glycolipid antigen, a-galactosylceramide (aGalCer), presented in the context of the non-classical antigen-presenting molecule, CD Id. However, NKT cells exposed to aGalCer can lyse a number of CD Id-negative tumor-derived ;ell lines. Lysis of these tumor targets in the absence of CDld suggests that aGalCer-activuted NKT cells may release soluble mediators. One possible mediator is granulysin, a newly described component of cytolytic granules present in activated cytotoxic T cells and NK cells. Granulysin is a member of the saposin-like protein family that has direct antimicrobial activity and that can activate caspase 9 to induce apoptosis in nucleated cells. In the present study, immunofluorescent staining demonstrated a granular pattern of granulysin expression in NKT cells. Purified granulysin protein was tested against a number of ta'get cells and correlated with results of cytotoxicity assays. The CDld-positive Molt-3 T cell acute lymphoblastic leukemia (ALL) cell line was specifically lysed by an NKT cell clone derived from a normal donor at the effector to target ratio of 5:1 (40{\%} specific lysis) and was sensitive to lysis by granulysin (30{\%} increase in propidium iodide-staining cells;. In contrast, a recently derived CD Id-negative T cell leukemia cell line was resistant to both lysis by NKT cells (3{\%} specific lysis) and to lysis by granulysin (10{\%} lysis). This cell line was established from a patient with chemo-refractory disease and contairs a chromosomal rearrangement at Ip36.1, the location of the caspase 9 gene. Caspase 9 is required for apoptosis induced by chemotherapeutic agents, as well as by granulysin. Loss of caspase 9 activity in this cell line is being investigated. Finally, primary ALL cells from an untreated patient were sensitive to granulysin (30{\%} lysis). Overall, these studies suggest that the tumorolytic effect of aGalCer-activated NKT cells may be mediated through the release of soluble cytotoxic mediators, such as granulysin, and that the optima) timing of immunotherapy using the NKT cell/aGalCer system is early in disease, before resistance emerges.",
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AU - Gansert, Jennifer L.

AU - Ochoa, Maria T.

AU - Krensky, Alan M.

AU - Porcelli, Steven A.

AU - Modlin, Robert L.

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