Blocked and non-blocked ricin immunotoxins against the CD4 antigen exhibit higher cytotoxic potency than a ricin A chain immunotoxin potentiated with ricin B chain or with a ricin B chain immunotoxin

Edward J. Wawrzynczak, Graham J. Watson, Alan J. Cumber, Raymond V. Henry, Geoffrey D. Parnell, E. Peter Rieber, Philip E. Thorpe

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

An immunotoxin consisting of ricin A chain linked to the monoclonal antibody M-T151, recognising the CD4 antigen, was weakly toxic to the human T-lymphoblastoid cell line CEM in tissue culture. The incorporation of [3H]leucine by CEM cells was inhibited by 50% at an M-T151-ricin-A-chain concentration (IC50) of 4.6 nM compared with an IC50 of 1.0 pM for ricin. In contrast, immunotoxins made by linking intact ricin to M-T151 in such a way that the galactose-binding sites of the B chain subunit were either blocked sterically by the antibody component or were left unblocked, were both powerfully cytotoxic with IC50 values of 20-30 pM. The addition of ricin B chain to CEM cells treated with M-T151-ricin-A-chain enhanced cytotoxicity by only eight-fold indicating that isolated B chain potentiated the action of the A chain less effectively than it did as an integral component of an intact ricin immunotoxin. Ricin B chain linked to goat anti-(mouse immunoglobulin) also potentiated weakly. Lactose completely inhibited the ability of isolated ricin B chain to potentiate the cytotoxicity of M-T151-ricin-A-chain and partially (3- to 4-fold) inhibited the cytotoxicity of the blocked and non-blocked ricin immunotoxins. Thus, in this system, the galactose-binding sites of the B chain contributed to cell killing regardless of whether isolated B chain was associated with the A chain immunotoxin or was present in blocked or non-blocked form as part of an intact ricin immunotoxin. The findings suggest that the blocked ricin immunotoxin may become unblocked after binding to the target antigen to re-expose the cryptic galactose-binding sites. However, the unblocking cannot be complete because the maximal inhibition of [3H]leucine incorporation by the blocked immunotoxin was only 80% compared with greater than 99% inhibition by the non-blocked immunotoxin.

Original languageEnglish (US)
Pages (from-to)289-295
Number of pages7
JournalCancer Immunology Immunotherapy
Volume32
Issue number5
DOIs
StatePublished - Sep 1991

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Ricin
Immunotoxins
CD4 Antigens
Galactose
Inhibitory Concentration 50
Binding Sites
Leucine
Poisons
Lactose
Goats
Immunoglobulins

ASJC Scopus subject areas

  • Oncology
  • Immunology
  • Cancer Research

Cite this

Blocked and non-blocked ricin immunotoxins against the CD4 antigen exhibit higher cytotoxic potency than a ricin A chain immunotoxin potentiated with ricin B chain or with a ricin B chain immunotoxin. / Wawrzynczak, Edward J.; Watson, Graham J.; Cumber, Alan J.; Henry, Raymond V.; Parnell, Geoffrey D.; Rieber, E. Peter; Thorpe, Philip E.

In: Cancer Immunology Immunotherapy, Vol. 32, No. 5, 09.1991, p. 289-295.

Research output: Contribution to journalArticle

Wawrzynczak, Edward J. ; Watson, Graham J. ; Cumber, Alan J. ; Henry, Raymond V. ; Parnell, Geoffrey D. ; Rieber, E. Peter ; Thorpe, Philip E. / Blocked and non-blocked ricin immunotoxins against the CD4 antigen exhibit higher cytotoxic potency than a ricin A chain immunotoxin potentiated with ricin B chain or with a ricin B chain immunotoxin. In: Cancer Immunology Immunotherapy. 1991 ; Vol. 32, No. 5. pp. 289-295.
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abstract = "An immunotoxin consisting of ricin A chain linked to the monoclonal antibody M-T151, recognising the CD4 antigen, was weakly toxic to the human T-lymphoblastoid cell line CEM in tissue culture. The incorporation of [3H]leucine by CEM cells was inhibited by 50{\%} at an M-T151-ricin-A-chain concentration (IC50) of 4.6 nM compared with an IC50 of 1.0 pM for ricin. In contrast, immunotoxins made by linking intact ricin to M-T151 in such a way that the galactose-binding sites of the B chain subunit were either blocked sterically by the antibody component or were left unblocked, were both powerfully cytotoxic with IC50 values of 20-30 pM. The addition of ricin B chain to CEM cells treated with M-T151-ricin-A-chain enhanced cytotoxicity by only eight-fold indicating that isolated B chain potentiated the action of the A chain less effectively than it did as an integral component of an intact ricin immunotoxin. Ricin B chain linked to goat anti-(mouse immunoglobulin) also potentiated weakly. Lactose completely inhibited the ability of isolated ricin B chain to potentiate the cytotoxicity of M-T151-ricin-A-chain and partially (3- to 4-fold) inhibited the cytotoxicity of the blocked and non-blocked ricin immunotoxins. Thus, in this system, the galactose-binding sites of the B chain contributed to cell killing regardless of whether isolated B chain was associated with the A chain immunotoxin or was present in blocked or non-blocked form as part of an intact ricin immunotoxin. The findings suggest that the blocked ricin immunotoxin may become unblocked after binding to the target antigen to re-expose the cryptic galactose-binding sites. However, the unblocking cannot be complete because the maximal inhibition of [3H]leucine incorporation by the blocked immunotoxin was only 80{\%} compared with greater than 99{\%} inhibition by the non-blocked immunotoxin.",
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T1 - Blocked and non-blocked ricin immunotoxins against the CD4 antigen exhibit higher cytotoxic potency than a ricin A chain immunotoxin potentiated with ricin B chain or with a ricin B chain immunotoxin

AU - Wawrzynczak, Edward J.

AU - Watson, Graham J.

AU - Cumber, Alan J.

AU - Henry, Raymond V.

AU - Parnell, Geoffrey D.

AU - Rieber, E. Peter

AU - Thorpe, Philip E.

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