High specific cytotoxicity of antibody-toxin hybrid molecules (immunotoxins) for target cells

Jansen, F. K.; Blythman, Hildur E.; Carrière, D.; Casellas, Pierre; Diaz, Juan E.; Gros, Pierre; Hennequin, J.R.; Paolucci, F.; Pau, Bernard; Poncelet, Pascal; Richer, G.; Salhi, Sharon Lynn; Vidal, Hubert; Voisin, G A

doi:10.1016/0165-2478(80)90057-7

Cited by 56 publications

(12 citation statements)

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“…Ricin A chain, on the other hand, when attached through a disulphide bridge to carrier molecules, can sometimes mediate selective cytotoxic effects with a potency similar to that of the native toxin. This result has been reported by Yamaguchi et al [28] using concanavalin A as the carrier and by Jansen and his co-workers [29,30] using monoclonal anti-dinitrophenyl and anti-Thyl .2 antibodies. Recently, Uchida et al [31] showed that rich A chain linked by a disulphide bond to a subunit of Wistaria jloribunda lectin was 100 -200 times more cytotoxic than a comparable conjugate with the A chain from diphtheria toxin.…”

Section: Discussionsupporting

confidence: 72%

“…Other workers have prepared conjugates of antibodies with A chains derived from bacterial [26,49] or plant toxins [29,49] and the conjugates have exhibited selective cytotoxicity in tissue culture, but their potencies have been variable. The advantage of using conjugates containing A chains or inhibitors like gelonin is that, lacking the capacity to bind non-specifically to cells, they should not be poisonous to animals.…”

mentioning

confidence: 99%

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Cytotoxicity Acquired by Conjugation of an Anti‐Thy_1.1 Monoclonal Antibody and the Ribosome‐Inactivating Protein, Gelonin

Thorpe

Brown

Ross

et al. 1981

European Journal of Biochemistry

117

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Gelonin, a plant protein which can powerfully reduce the protein-synthetic capacity of ribosome preparations, was covalently coupled to anti-Thy1 .I antibody. The conjugate was prepared using N-succinimidyl-3-(2-pyridyldithi0)propionate which generates a disulphide linkage between the component molecules. Two conjugate fractions were obtained with M, of 180000 and > 200000.After its linkage to the antibody, gelonin suppressed those Thy1.1-bearing T lymphocytes from AKR mice which will respond to phytohaemagglutinin and concanavalin A in tissue culture. The [3H]leucine incorporation with the T-cell mitogens was inhibited by 50% with the 180000-M, fraction at a concentration of 0.4nM and with the > 200000-Mr fraction of pM. Unconjugated gelonin induced comparable reductions in T-cell responsiveness but at concentrations of 30 nM. The conjugates exerted little or n o effect upon B lymphocytes or T lyniphocytes from CBA mice (Thyl.z+ve). Two Thyl.l-expressing AKR lymphoma cell lines, AKR-A and BW5147, were found to be sensitive to the conjugates, albeit much less so than the norinal T lymphocytes, The conjugates injected in vivo significantly prolonged the life of CBA mice bearing an AKR-A lymphoma allograft.It is concluded that gelonin can, by its linkage to an antibody, be rendered cytotoxic with a potency to match or exceed those of the toxins abrin and ricin.Gelonin, a glycoprotein from the seeds of Gelonium nzulti-,florum, inhibits protein synthesis in rabbit reticulocyte lysates by damaging the 60-S subunit of the ribosomes. The inactivation of the ribosomes is irreversible, seems not to involve cofactors, and occurs with an efficiency that suggests lhat gelonin acts enzymically [l]. Several other plants have been found to contain proteins which can inactivate isolated eukaryotic ribosomes apparently by the same mechanism as does gelonin. The proteins can be divided into those with and those without the capacity to recognise and bind to carbohydrates on cell surfaces.The inhibitors which d o not bind to cells are single polypeptide chains and are more or less devoid of toxic effects upon mammalian cells. This group, to which gelonin belongs, also includes the pokeweed antiviral protein [2,3] and inhibitors from wheat germ [4,5], Momordica charuntia [6] and probably also from Croton tiglium and Jatropha curcus [7].The proteins which bind to cells d o so because the ribosomc-inhibitory subunit (the A chain) is linked by a disulphide bond to a second subunit (the B chain) which has the cell-binding property. These proteins are either monovalent or, as with the lectins, divalent with two A-B units associated noncovalently. They differ in toxicity both to intact animals and to cells in tissue culture. Some, such as ricin and abrin (reviewed in [S]), modeccin [9,10] and a toxic lectin from Viscum album [Ill, are extremely potent toxins whereas others are less toxic or non-toxic, such as Ricinus communis agglutinin [12], Ahrus precatorius agglutinin [8,13] and thc lectins from iz4. charantia, Vicia craccu and Crotulu...

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Section: Discussionsupporting

confidence: 72%

mentioning

confidence: 99%

Cytotoxicity Acquired by Conjugation of an Anti‐Thy_1.1 Monoclonal Antibody and the Ribosome‐Inactivating Protein, Gelonin

Thorpe

Brown

Ross

et al. 1981

European Journal of Biochemistry

117

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“…When antibodies directed against surface determinants on a wide variety of cells are coupled to intact ricin and used at sufficient concentrations in vitro in the presence of galactose (to block the galactose-binding site of the B chain), virtually all cells expressing the relevant determinant can be killed (7)(8)(9)(10)(11)(12)(13). In addition, free B chains can synergize in vitro with A-chain-containing immunotoxins in killing specific target cells (5).…”

mentioning

confidence: 99%

Synergy of ricin A chain-containing immunotoxins and ricin B chain-containing immunotoxins in in vitro killing of neoplastic human B cells.

Vitetta¹,

Cushley²,

Uhr³

1983

Proc. Natl. Acad. Sci. U.S.A.

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A strategy is described for improving the efficacy of ricin A chain-containing immunotoxins. Highly purified preparations of ricin A chain or ricin B chain were separately coupled to anti-human immunoglobulin antibodies and the conjugates (immunotoxins) were affinity purified to eliminate free chains. Mixtures of anti-Ig-A and anti-Ig-B immunotoxins markedly synergized in vitro in their ability to kill the Ig-bearing human lymphoma cell line Daudi. In contrast, A-chain-or B-chain-containing immunotoxins of irrelevant specificity did not synergize with antiIg-A or anti-Ig-B immunotoxins. This finding indicates that free A or B chains do not play a major role in the synergy and that the synergy is specific. Thus, synergy depends on the specificity of the two antibodies; the lectin binding ability of a B-chain-containing immunotoxin of irrelevant specificity does not suffice. This approach of delivering ricin A chain and B chain separately to a target cell may have significant advantages in killing cells that are not effectively killed by A-chain-containing immunotoxins alone.The plant toxin ricin is composed of an A chain that is disulfide bonded to a B chain. The B chain binds to galactose-containing glycoproteins and glycolipids and hence to virtually all eukaryotic cells. The A chain can enzymatically inhibit protein synthesis after its entry into the cell cytoplasm. Antibodies directed against the isotype or idiotype of cell surface immunoglobulin of normal or neoplastic B lymphocytes, coupled to the A chain of ricin, can kill target cells both in vitro (1-3) and in vivo (1). Although such immunotoxins are highly effective in killing cells bearing the relevant surface Ig determinant(s), A-chain-containing immunotoxins directed against determinants on other types of cells (e.g., T cells) are frequently less effective (4-6).When antibodies directed against surface determinants on a wide variety of cells are coupled to intact ricin and used at sufficient concentrations in vitro in the presence of galactose (to block the galactose-binding site of the B chain), virtually all cells expressing the relevant determinant can be killed (7-13). In addition, free B chains can synergize in vitro with A-chain-containing immunotoxins in killing specific target cells (5). However, the in vivo use of ricin-containing immunotoxins is precluded by the nonspecific tissue toxicity conferred by the ricin B chain (14).It is postulated that the greater toxicity of ricin-containing immunotoxins, as compared with A-chain-containing immunotoxins, is due to a second function of the B chain-i.e., its ability to facilitate the entry of A chains into the cytoplasm in addition to its receptor binding function (reviewed in ref. 5). It would therefore be desirable to develop a strategy in which the putative transport role of the B chain could be preserved while its function as a lectin was eliminated or minimized. To this end, we have developed an approach that uses two types of immunotoxins. Antibodies reactive with human B lymphocyte...

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“…In addition, our previous study [19] on a conjugate of ricin A-chain with a monoclonal antibody to MM antigen, mouse mammary tumor-specific antigen, demonstrated in vivo antitumor efficacy of the conjugate against the MM-antigen-positive ascites mouse mammary tumor MM46. Other investigators have also studied the toxin A-chain (fragment) conjugates [1,7,8,10,14,17,18], but most of the studies have been limited to in vitro experiments or carried out using an antibody to T cell differentiation antigen, Thyl, as a model system or an antibody to an immunoglobulin idiotype.…”

Section: Introductionmentioning

confidence: 99%

Ricin A-chain conjugated with monoclonal anti-L1210 antibody

Kishida

Masuho

Saito

et al. 1983

Cancer Immunol Immunother

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In studies of antitumor antibody-cytotoxic agent conjugates as potential antitumor agents with improved tumor specificity, the toxic subunit A-chain of ricin was conjugated with a monoclonal antibody to a tumor-associated antigen expressed weakly on murine leukemia L1210 cells and strongly on L1210/GZL cells, a guanazole-resistant subline of L1210, employing N-succinimidyl 3-(2-pyridyldithio)propionate as cross-linking agent. The conjugate (anti-L1210 conjugate) exhibited a potent concentration-dependent cytotoxicity against cultured L1210/GZL cells, and inhibited cell growth at concentrations over 0.8 micrograms/ml. The conjugate killed all L1210/GZL cells at a concentration of 100 micrograms/ml. Neither nonimmune conjugate similarly prepared from mouse nonimmune IgG nor unconjugated anti-L1210 IgG alone showed cytotoxicity against L1210/GZL cells. When (BALB/c X DBA/2)F1 mice inoculated with 1 X 10(5) L1210/GZL cells were treated with IP injections of 27 micrograms anti-L1210 conjugate 1 h and 5 days after tumor cell inoculation, a life-prolonging effect was observed. [Lifespan in treated animals as percentage of that in controls (T/C) = 146%]. However, when the dose per injection was increased to 50 micrograms per mouse, survival was the same as in the control group. Postmortem examination of mice that had been treated with 50 micrograms anti-L1210 conjugate revealed lesions with necrosis and hemorrhage in the liver parenchyma and the intestinal epithelium, respectively. A similar toxic effect on the host mice was also observed with nonimmune conjugate.

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High specific cytotoxicity of antibody-toxin hybrid molecules (immunotoxins) for target cells

Cited by 56 publications

References 12 publications

Cytotoxicity Acquired by Conjugation of an Anti‐Thy_1.1 Monoclonal Antibody and the Ribosome‐Inactivating Protein, Gelonin

Cytotoxicity Acquired by Conjugation of an Anti‐Thy_1.1 Monoclonal Antibody and the Ribosome‐Inactivating Protein, Gelonin

Synergy of ricin A chain-containing immunotoxins and ricin B chain-containing immunotoxins in in vitro killing of neoplastic human B cells.

Ricin A-chain conjugated with monoclonal anti-L1210 antibody

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High specific cytotoxicity of antibody-toxin hybrid molecules (immunotoxins) for target cells

Cited by 56 publications

References 12 publications

Cytotoxicity Acquired by Conjugation of an Anti‐Thy1.1 Monoclonal Antibody and the Ribosome‐Inactivating Protein, Gelonin

Cytotoxicity Acquired by Conjugation of an Anti‐Thy1.1 Monoclonal Antibody and the Ribosome‐Inactivating Protein, Gelonin

Synergy of ricin A chain-containing immunotoxins and ricin B chain-containing immunotoxins in in vitro killing of neoplastic human B cells.

Ricin A-chain conjugated with monoclonal anti-L1210 antibody

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Product

Resources

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Cytotoxicity Acquired by Conjugation of an Anti‐Thy_1.1 Monoclonal Antibody and the Ribosome‐Inactivating Protein, Gelonin

Cytotoxicity Acquired by Conjugation of an Anti‐Thy_1.1 Monoclonal Antibody and the Ribosome‐Inactivating Protein, Gelonin