A disulfide conjugate between anti‐tetanus antibodies and HIV (37–72)Tat neutralizes tetanus toxin inside chromaffin cells

Abstract

Conjugates between anti-tetanus F(abP P) 2 fragments and the (37^72) fragment of the HIV Tat protein were taken up by chromaffin cells, NG108-15 neurohybridoma cells and Rev-2-T-6 lymphoma cells. The uptake could not be inhibited by competition with (37^72)Tat, but was reduced in the presence of metabolic inhibitors or at low temperature. The disulfide as well as the thioether conjugate were translocated to the cytoplasmic space, but only the disulfide conjugate moderately restored the stimulated transmitter release inhibited by tetanus toxin. Therefore, disulfide conjugates are more promising than thioethers for the neutralization of intracellular antigens. These conjugates provide new tools to study neuroprotection against bacterial neurotoxins.z 1999 Federation of European Biochemical Societies.

Cell-penetrating and cell-targeting peptides in drug delivery

“…They demonstrated that only the disulfide conjugates effectively neutralized the toxin. Surprisingly, the disulfide conjugate also showed a higher nuclear accumulation compared to the thioether conjugate [72]. One would have expected the opposite result since the intracellular reducing environment should be strong enough to cause the disulfide bridge reduction as shown in a previous study with a CPP-peptide conjugate [73].…”

“…Stein et al conjugated a longer version of the Tat peptide spanning along residues 37 to 72 of the HIV-1 Tat protein to a Fab fragment of anti-tetanus toxin antibodies either by thioether or disulfide linkage [72]. They demonstrated that only the disulfide conjugates effectively neutralized the toxin.…”

Targeting antibodies to the cytoplasm

“…Chemical conjugation was employed for Antibodies or antibody fragments linked to PTDs have been used to neutralize tetanus toxin in chromaffin cells and influenza A viral activity. 142,156 Further applications included interfering with the cell cycle, such as inhibition of a G 1 -S-phase arrest by an anti-Cdk (cyclin dependent kinase) inhibitor, 154 or inhibition of cell cycle progression by an anti-cyclin D1 transbody. 151 Transbodies have also been used to target apoptosis in order to promote, as well as to suppress, apoptosis.…”

Cell‐penetrating peptides: Nanocarrier for macromolecule delivery in living cells

“…Recombinant SOCS3-CPP has been shown to effectively inhibit the cytokine-mediated signal transduction associated with acute inflammation and liver apoptosis (67). Disulfide linkage between Tat and anti-tetanus antibody caused neutralization of the toxin and showed higher nuclear localization (68). Transduction of transposase sleeping beauty protein using M918 has been reported (69).…”