Antibodies have been used to protect against the effects of biological toxins for over a century. It is generally accepted that the mechanism whereby antibodies protect is by blocking the entry of toxins into cells. However, this ignores an important body of evidence demonstrating that antibodies to the A-chain of A-B toxins have equal or greater neutralizing activity than anti-B chain antibodies. We will present data demonstrating this with ricin toxin. Based upon these results we have formed the hypothesis that antibodies can protect by altering the intracellular processing and routing of the toxin and that the affinity of antibody binding may determine its ability to do so. To test this hypothesis we propose the following Specific Aims: ? ? Specific Aim 1: To produce and characterize a panel of high affinity anti-ricin A chain antibodies. The relationship between antibody affinity, in vitro neutralization and in vivo protection will be studied. ? ? Specific Aim 2: To study the intracellular routing of fluorescent-labeled ricin toxin in the presence of neutralizing and non-neutralizing antibodies. Confocal, deconvolution, and electron microscopy, and subcellular isolations will be used to study effects of antibody on subcellular localization of ricin. ? ? Specific Aim 3: To study the intracellular association between toxin and antibody. Double-label studies and fluorescence-energy transfer (FRET) will be used to determine how long antibody remains bound to the toxin during intracellular processing. ? ? Ricin is a prototype A-B toxin, a group that includes many bacterial and plant toxins. In addition to defining basic processes, the studies proposed in this application have utility for the development of vaccines and treatments for intoxications. ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI059376-04
Application #
7015595
Study Section
Special Emphasis Panel (ZRG1-VACC (02))
Program Officer
Schmitt, Clare K
Project Start
2004-02-01
Project End
2009-01-31
Budget Start
2006-02-01
Budget End
2007-01-31
Support Year
4
Fiscal Year
2006
Total Cost
$489,550
Indirect Cost
Name
Children's Hospital (New Orleans)
Department
Type
DUNS #
069523405
City
New Orleans
State
LA
Country
United States
Zip Code
70118
Pincus, Seth H; Das, Anushka; Song, Kejing et al. (2014) Role of Fc in antibody-mediated protection from ricin toxin. Toxins (Basel) 6:1512-25
Sun, Yong; Zhu, Yun; Wang, Liangxi et al. (2013) Recombinant adenovirus-mediated intestinal trefoil factor gene therapy for burn-induced intestinal mucosal injury. PLoS One 8:e62429
Song, Kejing; Mize, R Ranney; Marrero, Luis et al. (2013) Antibody to ricin a chain hinders intracellular routing of toxin and protects cells even after toxin has been internalized. PLoS One 8:e62417
Zhao, Zhiyu; Worthylake, David; LeCour Jr, Louis et al. (2012) Crystal structure and computational modeling of the fab fragment from a protective anti-ricin monoclonal antibody. PLoS One 7:e52613
Roy, Chad J; Song, Kejing; Sivasubramani, Satheesh K et al. (2012) Animal models of ricin toxicosis. Curr Top Microbiol Immunol 357:243-57
Pincus, Seth H; Smallshaw, Joan E; Song, Kejing et al. (2011) Passive and active vaccination strategies to prevent ricin poisoning. Toxins (Basel) 3:1163-84
Roche, James K; Stone, Matthew K; Gross, Lisa K et al. (2008) Post-exposure targeting of specific epitopes on ricin toxin abrogates toxin-induced hypoglycemia, hepatic injury, and lethality in a mouse model. Lab Invest 88:1178-91
Maddaloni, Massimo; Cooke, Corrie; Wilkinson, Royce et al. (2004) Immunological characteristics associated with the protective efficacy of antibodies to ricin. J Immunol 172:6221-8