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Neutralising antibodies against ricin toxin.

Prigent J, Panigai L, Lamourette P, Sauvaire D, Devilliers K, Plaisance M, Volland H, Créminon C, Simon S - PLoS ONE (2011)

Bottom Line: An immunotherapeutic approach could be of interest to attenuate or neutralise the effects of the toxin.We sought to characterise neutralising monoclonal antibodies against ricin and to develop an effective therapy.Seven mAbs were selected for their capacity to neutralise the cytotoxic effects of ricin in vitro.

View Article: PubMed Central - PubMed

Affiliation: CEA, iBiTec-S, Service de Pharmacologie et d'Immunoanalyse, CEA Saclay, Gif sur Yvette, France.

ABSTRACT
The Centers for Disease Control and Prevention have listed the potential bioweapon ricin as a Category B Agent. Ricin is a so-called A/B toxin produced by plants and is one of the deadliest molecules known. It is easy to prepare and no curative treatment is available. An immunotherapeutic approach could be of interest to attenuate or neutralise the effects of the toxin. We sought to characterise neutralising monoclonal antibodies against ricin and to develop an effective therapy. For this purpose, mouse monoclonal antibodies (mAbs) were produced against the two chains of ricin toxin (RTA and RTB). Seven mAbs were selected for their capacity to neutralise the cytotoxic effects of ricin in vitro. Three of these, two anti-RTB (RB34 and RB37) and one anti-RTA (RA36), when used in combination improved neutralising capacity in vitro with an IC(50) of 31 ng/ml. Passive administration of association of these three mixed mAbs (4.7 µg) protected mice from intranasal challenges with ricin (5 LD(50)). Among those three antibodies, anti-RTB antibodies protected mice more efficiently than the anti-RTA antibody. The combination of the three antibodies protected mice up to 7.5 hours after ricin challenge. The strong in vivo neutralising capacity of this three mAbs combination makes it potentially useful for immunotherapeutic purposes in the case of ricin poisoning or possibly for prevention.

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Related in: MedlinePlus

Immunoblot of ricin identified by several anti-ricin antibodies.2 µg of ricin was migrated in 15% SDS-PAGE and blotted onto PVDF membrane. Primary monoclonal antibodies obtained against the A or B chain were incubated for 1 h to bind to ricin. A secondary antibody HRP-conjugated anti-mouse IgG (diluted 1/2000) was added and proteins were detected after 10 min by chemiluminescence (ECL) using a VersaDoc imaging system (Bio-Rad). Two lanes are shown for each antibody, corresponding to the migration of ricin and of molecular weight markers (two lines, 37 and 20 kDa), respectively.
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pone-0020166-g001: Immunoblot of ricin identified by several anti-ricin antibodies.2 µg of ricin was migrated in 15% SDS-PAGE and blotted onto PVDF membrane. Primary monoclonal antibodies obtained against the A or B chain were incubated for 1 h to bind to ricin. A secondary antibody HRP-conjugated anti-mouse IgG (diluted 1/2000) was added and proteins were detected after 10 min by chemiluminescence (ECL) using a VersaDoc imaging system (Bio-Rad). Two lanes are shown for each antibody, corresponding to the migration of ricin and of molecular weight markers (two lines, 37 and 20 kDa), respectively.

Mentions: The 20 anti-RTB and 11 anti-RTA antibodies were tested for their capacity to recognise denatured ricin in reducing conditions in SDS-PAGE/western blotting analysis (Fig. 1). Four out of the 31 antibodies bound the denatured protein and thus possibly a linear epitope during immunoblot experiments (RA31 RA33, RA35 and RB37, Fig. 1).


Neutralising antibodies against ricin toxin.

Prigent J, Panigai L, Lamourette P, Sauvaire D, Devilliers K, Plaisance M, Volland H, Créminon C, Simon S - PLoS ONE (2011)

Immunoblot of ricin identified by several anti-ricin antibodies.2 µg of ricin was migrated in 15% SDS-PAGE and blotted onto PVDF membrane. Primary monoclonal antibodies obtained against the A or B chain were incubated for 1 h to bind to ricin. A secondary antibody HRP-conjugated anti-mouse IgG (diluted 1/2000) was added and proteins were detected after 10 min by chemiluminescence (ECL) using a VersaDoc imaging system (Bio-Rad). Two lanes are shown for each antibody, corresponding to the migration of ricin and of molecular weight markers (two lines, 37 and 20 kDa), respectively.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3102095&req=5

pone-0020166-g001: Immunoblot of ricin identified by several anti-ricin antibodies.2 µg of ricin was migrated in 15% SDS-PAGE and blotted onto PVDF membrane. Primary monoclonal antibodies obtained against the A or B chain were incubated for 1 h to bind to ricin. A secondary antibody HRP-conjugated anti-mouse IgG (diluted 1/2000) was added and proteins were detected after 10 min by chemiluminescence (ECL) using a VersaDoc imaging system (Bio-Rad). Two lanes are shown for each antibody, corresponding to the migration of ricin and of molecular weight markers (two lines, 37 and 20 kDa), respectively.
Mentions: The 20 anti-RTB and 11 anti-RTA antibodies were tested for their capacity to recognise denatured ricin in reducing conditions in SDS-PAGE/western blotting analysis (Fig. 1). Four out of the 31 antibodies bound the denatured protein and thus possibly a linear epitope during immunoblot experiments (RA31 RA33, RA35 and RB37, Fig. 1).

Bottom Line: An immunotherapeutic approach could be of interest to attenuate or neutralise the effects of the toxin.We sought to characterise neutralising monoclonal antibodies against ricin and to develop an effective therapy.Seven mAbs were selected for their capacity to neutralise the cytotoxic effects of ricin in vitro.

View Article: PubMed Central - PubMed

Affiliation: CEA, iBiTec-S, Service de Pharmacologie et d'Immunoanalyse, CEA Saclay, Gif sur Yvette, France.

ABSTRACT
The Centers for Disease Control and Prevention have listed the potential bioweapon ricin as a Category B Agent. Ricin is a so-called A/B toxin produced by plants and is one of the deadliest molecules known. It is easy to prepare and no curative treatment is available. An immunotherapeutic approach could be of interest to attenuate or neutralise the effects of the toxin. We sought to characterise neutralising monoclonal antibodies against ricin and to develop an effective therapy. For this purpose, mouse monoclonal antibodies (mAbs) were produced against the two chains of ricin toxin (RTA and RTB). Seven mAbs were selected for their capacity to neutralise the cytotoxic effects of ricin in vitro. Three of these, two anti-RTB (RB34 and RB37) and one anti-RTA (RA36), when used in combination improved neutralising capacity in vitro with an IC(50) of 31 ng/ml. Passive administration of association of these three mixed mAbs (4.7 µg) protected mice from intranasal challenges with ricin (5 LD(50)). Among those three antibodies, anti-RTB antibodies protected mice more efficiently than the anti-RTA antibody. The combination of the three antibodies protected mice up to 7.5 hours after ricin challenge. The strong in vivo neutralising capacity of this three mAbs combination makes it potentially useful for immunotherapeutic purposes in the case of ricin poisoning or possibly for prevention.

Show MeSH
Related in: MedlinePlus