A recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells

Matthias Peipp, Heide Küpers, Domenica Saul, Beate Schlierf, Johann Greil, Susan J. Zunino, Martin Gramatzki, Georg H. Fey

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

A recombinant immunotoxin was constructed from the hybridoma antibody TH-69 directed against human CD7, a surface antigen of leukemic T cells. The antibody was subcloned as a single chain Fv (scFv) fragment and genetically linked to a truncated Pseudomonas exotoxin A fragment containing the catalytic domains II and III but lacking the receptor binding domain I. Domain I was replaced by the scFv, thus conferring restricted specificity for CD7.positive cells. The bacterially expressed and purified toxin retained binding specificity for CD7-positive cells. It promoted apoptosis in two CD7-positive cell lines derived from T-lineage acute lymphoblastic leukemias, CEM and Jurkat, but not in the CD7-negative B-lymphoid lines REH, Nalm-6, and SEM. Maximum killing in excess of 95% was reached after 96 h in CEM and Jurkat cells with a single dose of 100 ng/ml. Cells treated with a similarly constructed scFv-exotoxin A immunotoxin against melanoma-associated chondroitin sulfate proteoglycan, an antigen absent from leukemic T cells, remained unaffected. Lysis of target cells occurred via apoptosis as evidenced by staining with Annexin V and specific cleavage of poly(ADP-ribose) polymerase. Approximately 20% of leukemic cells from a patient with CD7-positive acute T-cell leukemia kept in long-term primary culture for 30 cell generations were killed within 96 h after treatment with the toxin. These findings justify further evaluation of the agent in view of potential therapeutic applications.

Original languageEnglish (US)
Pages (from-to)2848-2855
Number of pages8
JournalCancer Research
Volume62
Issue number10
StatePublished - May 15 2002
Externally publishedYes

Fingerprint

Immunotoxins
Single-Chain Antibodies
Apoptosis
T-Lymphocytes
Immunoglobulin Variable Region
Chondroitin Sulfate Proteoglycans
Precursor T-Cell Lymphoblastic Leukemia-Lymphoma
Exotoxins
Jurkat Cells
Primary Cell Culture
Poly(ADP-ribose) Polymerases
Antibodies
Annexin A5
Hybridomas
Surface Antigens
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Melanoma
Catalytic Domain
Staining and Labeling
Antigens

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Peipp, M., Küpers, H., Saul, D., Schlierf, B., Greil, J., Zunino, S. J., ... Fey, G. H. (2002). A recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells. Cancer Research, 62(10), 2848-2855.

A recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells. / Peipp, Matthias; Küpers, Heide; Saul, Domenica; Schlierf, Beate; Greil, Johann; Zunino, Susan J.; Gramatzki, Martin; Fey, Georg H.

In: Cancer Research, Vol. 62, No. 10, 15.05.2002, p. 2848-2855.

Research output: Contribution to journalArticle

Peipp, M, Küpers, H, Saul, D, Schlierf, B, Greil, J, Zunino, SJ, Gramatzki, M & Fey, GH 2002, 'A recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells', Cancer Research, vol. 62, no. 10, pp. 2848-2855.
Peipp M, Küpers H, Saul D, Schlierf B, Greil J, Zunino SJ et al. A recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells. Cancer Research. 2002 May 15;62(10):2848-2855.
Peipp, Matthias ; Küpers, Heide ; Saul, Domenica ; Schlierf, Beate ; Greil, Johann ; Zunino, Susan J. ; Gramatzki, Martin ; Fey, Georg H. / A recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells. In: Cancer Research. 2002 ; Vol. 62, No. 10. pp. 2848-2855.
@article{584d45606ae84cb8bb9d0deba489d9a6,
title = "A recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells",
abstract = "A recombinant immunotoxin was constructed from the hybridoma antibody TH-69 directed against human CD7, a surface antigen of leukemic T cells. The antibody was subcloned as a single chain Fv (scFv) fragment and genetically linked to a truncated Pseudomonas exotoxin A fragment containing the catalytic domains II and III but lacking the receptor binding domain I. Domain I was replaced by the scFv, thus conferring restricted specificity for CD7.positive cells. The bacterially expressed and purified toxin retained binding specificity for CD7-positive cells. It promoted apoptosis in two CD7-positive cell lines derived from T-lineage acute lymphoblastic leukemias, CEM and Jurkat, but not in the CD7-negative B-lymphoid lines REH, Nalm-6, and SEM. Maximum killing in excess of 95{\%} was reached after 96 h in CEM and Jurkat cells with a single dose of 100 ng/ml. Cells treated with a similarly constructed scFv-exotoxin A immunotoxin against melanoma-associated chondroitin sulfate proteoglycan, an antigen absent from leukemic T cells, remained unaffected. Lysis of target cells occurred via apoptosis as evidenced by staining with Annexin V and specific cleavage of poly(ADP-ribose) polymerase. Approximately 20{\%} of leukemic cells from a patient with CD7-positive acute T-cell leukemia kept in long-term primary culture for 30 cell generations were killed within 96 h after treatment with the toxin. These findings justify further evaluation of the agent in view of potential therapeutic applications.",
author = "Matthias Peipp and Heide K{\"u}pers and Domenica Saul and Beate Schlierf and Johann Greil and Zunino, {Susan J.} and Martin Gramatzki and Fey, {Georg H.}",
year = "2002",
month = "5",
day = "15",
language = "English (US)",
volume = "62",
pages = "2848--2855",
journal = "Journal of Cancer Research",
issn = "0099-7013",
publisher = "American Association for Cancer Research Inc.",
number = "10",

}

TY - JOUR

T1 - A recombinant CD7-specific single-chain immunotoxin is a potent inducer of apoptosis in acute leukemic T cells

AU - Peipp, Matthias

AU - Küpers, Heide

AU - Saul, Domenica

AU - Schlierf, Beate

AU - Greil, Johann

AU - Zunino, Susan J.

AU - Gramatzki, Martin

AU - Fey, Georg H.

PY - 2002/5/15

Y1 - 2002/5/15

N2 - A recombinant immunotoxin was constructed from the hybridoma antibody TH-69 directed against human CD7, a surface antigen of leukemic T cells. The antibody was subcloned as a single chain Fv (scFv) fragment and genetically linked to a truncated Pseudomonas exotoxin A fragment containing the catalytic domains II and III but lacking the receptor binding domain I. Domain I was replaced by the scFv, thus conferring restricted specificity for CD7.positive cells. The bacterially expressed and purified toxin retained binding specificity for CD7-positive cells. It promoted apoptosis in two CD7-positive cell lines derived from T-lineage acute lymphoblastic leukemias, CEM and Jurkat, but not in the CD7-negative B-lymphoid lines REH, Nalm-6, and SEM. Maximum killing in excess of 95% was reached after 96 h in CEM and Jurkat cells with a single dose of 100 ng/ml. Cells treated with a similarly constructed scFv-exotoxin A immunotoxin against melanoma-associated chondroitin sulfate proteoglycan, an antigen absent from leukemic T cells, remained unaffected. Lysis of target cells occurred via apoptosis as evidenced by staining with Annexin V and specific cleavage of poly(ADP-ribose) polymerase. Approximately 20% of leukemic cells from a patient with CD7-positive acute T-cell leukemia kept in long-term primary culture for 30 cell generations were killed within 96 h after treatment with the toxin. These findings justify further evaluation of the agent in view of potential therapeutic applications.

AB - A recombinant immunotoxin was constructed from the hybridoma antibody TH-69 directed against human CD7, a surface antigen of leukemic T cells. The antibody was subcloned as a single chain Fv (scFv) fragment and genetically linked to a truncated Pseudomonas exotoxin A fragment containing the catalytic domains II and III but lacking the receptor binding domain I. Domain I was replaced by the scFv, thus conferring restricted specificity for CD7.positive cells. The bacterially expressed and purified toxin retained binding specificity for CD7-positive cells. It promoted apoptosis in two CD7-positive cell lines derived from T-lineage acute lymphoblastic leukemias, CEM and Jurkat, but not in the CD7-negative B-lymphoid lines REH, Nalm-6, and SEM. Maximum killing in excess of 95% was reached after 96 h in CEM and Jurkat cells with a single dose of 100 ng/ml. Cells treated with a similarly constructed scFv-exotoxin A immunotoxin against melanoma-associated chondroitin sulfate proteoglycan, an antigen absent from leukemic T cells, remained unaffected. Lysis of target cells occurred via apoptosis as evidenced by staining with Annexin V and specific cleavage of poly(ADP-ribose) polymerase. Approximately 20% of leukemic cells from a patient with CD7-positive acute T-cell leukemia kept in long-term primary culture for 30 cell generations were killed within 96 h after treatment with the toxin. These findings justify further evaluation of the agent in view of potential therapeutic applications.

UR - http://www.scopus.com/inward/record.url?scp=0037093199&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037093199&partnerID=8YFLogxK

M3 - Article

C2 - 12019163

AN - SCOPUS:0037093199

VL - 62

SP - 2848

EP - 2855

JO - Journal of Cancer Research

JF - Journal of Cancer Research

SN - 0099-7013

IS - 10

ER -