A comparison of an anti-CD25 immunotoxin, Ontak and anti-CD25 microbeads for their ability to deplete alloreactive T cells in vitro

P. Vaclavkova, Y. Cao, L. K. Wu, J. Michalek, Ellen S. Vitetta

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Ex vivo depletion of alloreactive CD25+ T cells from a stem cell transplant (SCT) can reduce the incidence of graft-versus-host disease (GVHD) while preserving antimicrobial and perhaps antileukemia activity. However, the most effective methods for allodepleting T cells prior to transplant have not been determined. In this study, we have compared three agents that deplete CD25+ activated, alloreactive T cells. These included Ontak (Denileukin Diftitox), an IL-2 fusion toxin, anti-CD25 microbeads (MACS), an anti-CD25 immunotoxin (IT) and a combination of the IT and MACS. Peripheral blood mononuclear cells (PBMCs) activated in a primary mixed lymphocyte reaction (MLR) were allodepleted using optimal amounts of each agent, and the cells were then analyzed by flow cytometry. The treated cells were examined both for remaining alloreactivity and for the preservation of third party reactivity by testing them in a secondary MLR. Our data demonstrate that both the anti-CD25 IT and the anti-CD25 MACS were equally effective in depleting CD4+CD25+ cells and in sparing T cells that were reactive with third party cells. The anti-CD25 IT was, however, superior in depleting alloreactive CD8+CD25+ cells. In contrast, Ontak did not eliminate alloreactive cells and the Ontak-treated cells retained significant reactivity against the original stimulator cells.

Original languageEnglish (US)
Pages (from-to)559-567
Number of pages9
JournalBone Marrow Transplantation
Volume37
Issue number6
DOIs
StatePublished - Mar 2006

Fingerprint

Immunotoxins
Microspheres
T-Lymphocytes
Mixed Lymphocyte Culture Test
Transplants
denileukin diftitox
In Vitro Techniques
Graft vs Host Disease
Interleukin-2
Blood Cells
Flow Cytometry
Stem Cells
Incidence

Keywords

  • Alloreactive T cells
  • Anti-CD25 immunotoxin
  • Graft-versus-host disease
  • Magnetic cell sorting
  • Ontak
  • Selective depletion

ASJC Scopus subject areas

  • Hematology
  • Transplantation

Cite this

A comparison of an anti-CD25 immunotoxin, Ontak and anti-CD25 microbeads for their ability to deplete alloreactive T cells in vitro. / Vaclavkova, P.; Cao, Y.; Wu, L. K.; Michalek, J.; Vitetta, Ellen S.

In: Bone Marrow Transplantation, Vol. 37, No. 6, 03.2006, p. 559-567.

Research output: Contribution to journalArticle

@article{fdbb2f1255cf4fed9448d38a08864cc9,
title = "A comparison of an anti-CD25 immunotoxin, Ontak and anti-CD25 microbeads for their ability to deplete alloreactive T cells in vitro",
abstract = "Ex vivo depletion of alloreactive CD25+ T cells from a stem cell transplant (SCT) can reduce the incidence of graft-versus-host disease (GVHD) while preserving antimicrobial and perhaps antileukemia activity. However, the most effective methods for allodepleting T cells prior to transplant have not been determined. In this study, we have compared three agents that deplete CD25+ activated, alloreactive T cells. These included Ontak (Denileukin Diftitox), an IL-2 fusion toxin, anti-CD25 microbeads (MACS), an anti-CD25 immunotoxin (IT) and a combination of the IT and MACS. Peripheral blood mononuclear cells (PBMCs) activated in a primary mixed lymphocyte reaction (MLR) were allodepleted using optimal amounts of each agent, and the cells were then analyzed by flow cytometry. The treated cells were examined both for remaining alloreactivity and for the preservation of third party reactivity by testing them in a secondary MLR. Our data demonstrate that both the anti-CD25 IT and the anti-CD25 MACS were equally effective in depleting CD4+CD25+ cells and in sparing T cells that were reactive with third party cells. The anti-CD25 IT was, however, superior in depleting alloreactive CD8+CD25+ cells. In contrast, Ontak did not eliminate alloreactive cells and the Ontak-treated cells retained significant reactivity against the original stimulator cells.",
keywords = "Alloreactive T cells, Anti-CD25 immunotoxin, Graft-versus-host disease, Magnetic cell sorting, Ontak, Selective depletion",
author = "P. Vaclavkova and Y. Cao and Wu, {L. K.} and J. Michalek and Vitetta, {Ellen S.}",
year = "2006",
month = "3",
doi = "10.1038/sj.bmt.1705286",
language = "English (US)",
volume = "37",
pages = "559--567",
journal = "Bone Marrow Transplantation",
issn = "0268-3369",
publisher = "Nature Publishing Group",
number = "6",

}

TY - JOUR

T1 - A comparison of an anti-CD25 immunotoxin, Ontak and anti-CD25 microbeads for their ability to deplete alloreactive T cells in vitro

AU - Vaclavkova, P.

AU - Cao, Y.

AU - Wu, L. K.

AU - Michalek, J.

AU - Vitetta, Ellen S.

PY - 2006/3

Y1 - 2006/3

N2 - Ex vivo depletion of alloreactive CD25+ T cells from a stem cell transplant (SCT) can reduce the incidence of graft-versus-host disease (GVHD) while preserving antimicrobial and perhaps antileukemia activity. However, the most effective methods for allodepleting T cells prior to transplant have not been determined. In this study, we have compared three agents that deplete CD25+ activated, alloreactive T cells. These included Ontak (Denileukin Diftitox), an IL-2 fusion toxin, anti-CD25 microbeads (MACS), an anti-CD25 immunotoxin (IT) and a combination of the IT and MACS. Peripheral blood mononuclear cells (PBMCs) activated in a primary mixed lymphocyte reaction (MLR) were allodepleted using optimal amounts of each agent, and the cells were then analyzed by flow cytometry. The treated cells were examined both for remaining alloreactivity and for the preservation of third party reactivity by testing them in a secondary MLR. Our data demonstrate that both the anti-CD25 IT and the anti-CD25 MACS were equally effective in depleting CD4+CD25+ cells and in sparing T cells that were reactive with third party cells. The anti-CD25 IT was, however, superior in depleting alloreactive CD8+CD25+ cells. In contrast, Ontak did not eliminate alloreactive cells and the Ontak-treated cells retained significant reactivity against the original stimulator cells.

AB - Ex vivo depletion of alloreactive CD25+ T cells from a stem cell transplant (SCT) can reduce the incidence of graft-versus-host disease (GVHD) while preserving antimicrobial and perhaps antileukemia activity. However, the most effective methods for allodepleting T cells prior to transplant have not been determined. In this study, we have compared three agents that deplete CD25+ activated, alloreactive T cells. These included Ontak (Denileukin Diftitox), an IL-2 fusion toxin, anti-CD25 microbeads (MACS), an anti-CD25 immunotoxin (IT) and a combination of the IT and MACS. Peripheral blood mononuclear cells (PBMCs) activated in a primary mixed lymphocyte reaction (MLR) were allodepleted using optimal amounts of each agent, and the cells were then analyzed by flow cytometry. The treated cells were examined both for remaining alloreactivity and for the preservation of third party reactivity by testing them in a secondary MLR. Our data demonstrate that both the anti-CD25 IT and the anti-CD25 MACS were equally effective in depleting CD4+CD25+ cells and in sparing T cells that were reactive with third party cells. The anti-CD25 IT was, however, superior in depleting alloreactive CD8+CD25+ cells. In contrast, Ontak did not eliminate alloreactive cells and the Ontak-treated cells retained significant reactivity against the original stimulator cells.

KW - Alloreactive T cells

KW - Anti-CD25 immunotoxin

KW - Graft-versus-host disease

KW - Magnetic cell sorting

KW - Ontak

KW - Selective depletion

UR - http://www.scopus.com/inward/record.url?scp=33644891348&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33644891348&partnerID=8YFLogxK

U2 - 10.1038/sj.bmt.1705286

DO - 10.1038/sj.bmt.1705286

M3 - Article

VL - 37

SP - 559

EP - 567

JO - Bone Marrow Transplantation

JF - Bone Marrow Transplantation

SN - 0268-3369

IS - 6

ER -