Antigen binding and cytotoxic properties of a recombinant immunotoxin incorporating the lytic peptide, melittin

Dunn, Rosanne; Weston, Kathryn M; Longhurst, Terrence J.; Lilley, Glenn G.; Rivett, Donald E.; Hudson, Peter J.; Raison, Robert L.

doi:10.1016/s1380-2933(96)00055-3

Cited by 27 publications

(23 citation statements)

References 30 publications

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“…A melittin-based recombinant immunotoxin exhibited specific cytotoxicity against human tumour cells (Dunn et al, 1996) and more recently an HPMA copolymer-MLT conjugate showed in vitro toxicity towards B16F10 cells with reduced haemolytic activity (Musila and Duncan, 2001). This prompted consideration of ISA23-MLT as an anticancer polymer therapeutic.…”

Section: Discussionmentioning

confidence: 99%

Synthesis and preliminary evaluation of poly(amidoamine)–melittin conjugates as endosomolytic polymers and/or potential anticancer therapeutics

Lavignac

Lazenby

Franchini

et al. 2005

International Journal of Pharmaceutics

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The pH-responsive poly(amidoamine)s (PAAs) have been previously described. Whereas ISA23 enhances transfection in vitro and ISA1 promotes the cytosolic delivery of the non-permeant toxins this process shows poor efficiency. The aim of this study was to prepare and evaluate PAA conjugates containing the membrane disrupting peptide melittin (MLT). It was hypothesised that PAA conjugation would reduce the haemolytic activity of MLT at pH 7.4, however, upon delivery to tumours by the EPR effect, the polymer would uncoil in an acidic environment exposing MLT and allowing it to interact with membranes. PAA-MLT conjugates were prepared using MLT as a comonomer together with bis-acryloylpiperazine, 2-methylpiperazine and bis-hydroxyethylethylenediamine (ISA1-like), or bis-acrylamidoacetic acid and 2-methylpiperazine (ISA23-like). The melittin content of the conjugates was 6-19% (w/w). Although ISA1-MLT improved gelonin delivery compared to the parent polymer ISA1 (␣ 13-fold increase) and showed pH-dependent haemolytic activity at a polymer concentration of 0.05 mg/ml, this conjugate also displayed high haemolytic activity at pH 7.4. In contrast, ISA23-MLT like the parent compound ISA23 did not deliver gelonin. However, this conjugate could have potential as a novel polymeric anticancer conjugate due to its lack of haemolytic activity at pH 7.4 and retention of cytotoxicity.

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Section: Discussionmentioning

confidence: 99%

Synthesis and preliminary evaluation of poly(amidoamine)–melittin conjugates as endosomolytic polymers and/or potential anticancer therapeutics

Lavignac

Lazenby

Franchini

et al. 2005

International Journal of Pharmaceutics

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“…We have previously shown that a recombinant immunotoxin utilising melittin as the toxic component specifically bound to and killed target cells [Dunn et al, 1996]. In this case each immunotoxin molecule incorporated a single melittin molecule.…”

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confidence: 99%

Interaction of melittin with a human lymphoblastoid cell line, HMy2

Weston

Raison

1998

J. Cell. Biochem.

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We have examined the cytolytic effects of the membrane-active peptide, melittin, on a human lymphoblastoid cell line (HMy2) in the context of the use of melittin as the toxic component of an immunotoxin. The toxicity of melittin for HMy2 cells was linear over the concentration range 0.875-3.5 microM. Increased incubation times failed to result in significant cell death at concentrations of melittin below 0.875 microM. Kinetic analysis revealed that the cytolytic activity of melittin was independent of time of exposure beyond 90 min. Flow cytometric analysis of HMy2 cells incubated with FITC-labeled melittin demonstrated that the cells could incorporate up to 2.5 x 10(5) FITC-melittin molecules per cell with no reduction in viability. Extrapolation of this data indicates that 10(6) melittin molecules per cell are required for maximum cytotoxicity to HMy2 cells. Further analysis of HMy2 cells that incorporated melittin, but that remained viable, revealed that these cells were able to reduce the number of melittin molecules per cell over time. The data indicate a potential threshold value for the number of melittin molecules that may be required to be delivered to the cell surface in the form of an immunotoxin if effective selective cell death is to be achieved.

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“…Such molecules are small and highly cytotoxic, and they do not have to be internalized by a cell to kill it. This activity has recently led to a novel and potentially clinically important use of melittin as part of a recombinant immunotoxin (8). The immunotoxin retained the specificity of the original antibody and its cytotoxic activity, when tested against the specific cellular target in vitro, was significantly greater on a molar basis than that of native melittin, although the kinetics of killing were very different.…”

Section: Discussionmentioning

confidence: 99%

“…Residues 1-20 are able to form an amphiphilic ␣-helix and are important for binding to the cell membrane; the carboxyterminal hexapeptide contains four basic residues and seems to be vital for cytolysis (3,19). In recent years, the cytotoxic effects of melittin on human cell lines have been studied using flow cytometry (8,14,15,23). The criterion for cell death was the uptake (and consequent fluorescence) of either ethidium bromide or propidium iodide.…”

mentioning

confidence: 99%

Flow cytometric analysis of cell killing by the jumper ant venom peptide pilosulin 1

King

Donovan

et al. 1998

Cytometry

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Antigen binding and cytotoxic properties of a recombinant immunotoxin incorporating the lytic peptide, melittin

Cited by 27 publications

References 30 publications

Synthesis and preliminary evaluation of poly(amidoamine)–melittin conjugates as endosomolytic polymers and/or potential anticancer therapeutics

Synthesis and preliminary evaluation of poly(amidoamine)–melittin conjugates as endosomolytic polymers and/or potential anticancer therapeutics

Interaction of melittin with a human lymphoblastoid cell line, HMy2

Flow cytometric analysis of cell killing by the jumper ant venom peptide pilosulin 1

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