Specific ex-vivo depletion of human bone marrow Τ lymphocytes by an anti-pan-t cell (CD5) ricin A-chain immunotoxin

Salvatore Siena, Silvia Villa, Gianni Bonadonna, Marco Bregni, A. Massimo Gianni

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Abstract

We studied optimal conditions for ex vivo elimination of mature T cells from human bone marrow by T101 immunotoxin (T101-IT) with criteria applicable to graft-versus-host disease (GVHD) prophylaxis prior to allogeneic marrow transplantation. T101-IT consisted of T101 anti-CD5 monoclonal antibody conjugated to purified ricin A-chain toxin. Marrow mononuclear cells isolated by Ficoll-Hypaque or by fractionation with soybean lectin (SBA- cells) were incubated with T101-IT at 37°C with or without ammonium chloride and/or verapamil as potential enhancers of immunotoxin potency. As controls, competitive inhibition studies with unconjugated T101 or irrelevant IgG2a antibody were carried out. Residual T cells were quantified by limiting dilution in phytohemagglutinin (PHA)-interleukin 2 (IL-2) feeder-cell-containing microcultures and hematopoietic progenitors by CFU-GM assay. We demonstrated that T101-IT in the range of 1-100 nM does not affect early total cell viability; that its delayed cytotoxicity is T-cell-specific, greatly enhanced by ammonium chloride, and moderately by verapamil—which also is not synergistic with ammonium chloride; and that 10 nM × 3 fractionated doses (i.e., added at 0, 1.5, and 3 hr of incubation) in the presence of 10 mM ammonium chloride for 4 hr at pH 7.8 consistently induces 2 log T cell depletion. In addition, if the same T101-IT treatment is preceded by fractionation with soybean lectin (i.e., T101-IT treatment of SBA" marrow cells), 3 log T cell depletion is accomplished. We conclude that T101-IT is highly effective in eliminating T cells from donor grafts. However, data presented here indicate that T101-IT should be associated with additional methods, such as soybean lectin fractionation, to ensure more effective ex vivo T cell depletion and acute GVHD prevention.

Original languageEnglish
Pages (from-to)421-426
Number of pages6
JournalTransplantation
Volume43
Issue number3
Publication statusPublished - 1987

Fingerprint

Ricin
Immunotoxins
Bone Marrow
Lymphocytes
Ammonium Chloride
T-Lymphocytes
Graft vs Host Disease
Diatrizoate
Feeder Cells
Ficoll
Granulocyte-Macrophage Progenitor Cells
Homologous Transplantation
Phytohemagglutinins
Verapamil
Interleukin-2
Cell Survival
Monoclonal Antibodies
Transplants

ASJC Scopus subject areas

  • Transplantation
  • Immunology

Cite this

Siena, S., Villa, S., Bonadonna, G., Bregni, M., & Gianni, A. M. (1987). Specific ex-vivo depletion of human bone marrow Τ lymphocytes by an anti-pan-t cell (CD5) ricin A-chain immunotoxin. Transplantation, 43(3), 421-426.

Specific ex-vivo depletion of human bone marrow Τ lymphocytes by an anti-pan-t cell (CD5) ricin A-chain immunotoxin. / Siena, Salvatore; Villa, Silvia; Bonadonna, Gianni; Bregni, Marco; Gianni, A. Massimo.

In: Transplantation, Vol. 43, No. 3, 1987, p. 421-426.

Research output: Contribution to journalArticle

Siena, S, Villa, S, Bonadonna, G, Bregni, M & Gianni, AM 1987, 'Specific ex-vivo depletion of human bone marrow Τ lymphocytes by an anti-pan-t cell (CD5) ricin A-chain immunotoxin', Transplantation, vol. 43, no. 3, pp. 421-426.
Siena, Salvatore ; Villa, Silvia ; Bonadonna, Gianni ; Bregni, Marco ; Gianni, A. Massimo. / Specific ex-vivo depletion of human bone marrow Τ lymphocytes by an anti-pan-t cell (CD5) ricin A-chain immunotoxin. In: Transplantation. 1987 ; Vol. 43, No. 3. pp. 421-426.
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