Botulinum neurotoxin (BoNT) is an extremely potent neuromuscular blocker that has been designated a category A select bioterror agent by the United States Government because of its extreme lethality and its availability from environmental sources. Current countermeasures for BoNT exposure are insufficient, and there is an urgent need for therapeutic antibodies able to counteract the effects of BoNT exposure. Our working hypothesis is that a combination of human antibodies that bind BoNT will potently protect individuals from the complications of BoNT exposure. We have recently developed a novel hybridoma protocol for the cloning of human antibodies and have used it to clone antibodies that bind BoNT serotypes A and B. One of the serotype A antibodies is neutralizing in vivo. We plan to use our method to clone native human antibodies that bind and neutralize botulinum toxin serotypes A and B. For this purpose we will obtain peripheral blood lymphocytes from individuals vaccinated with the botulinum toxoid vaccine. We will assess in vitro and in vivo neutralizing activities of the cloned antibodies, singly and in combination. We will characterize the binding and primary sequences of the antibodies to determine the features that correlate with the ability to neutralize toxin and to cooperate synergistically in toxin neutralization. The primary result of these experiments will be the creation of combinations of cloned human antibodies that can effectively counteract BoNT/A and BoNT/B exposure. ? ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI065967-02
Application #
7176215
Study Section
Special Emphasis Panel (ZRG1-IDM-H (10))
Program Officer
Van de Verg, Lillian L
Project Start
2006-02-15
Project End
2007-11-30
Budget Start
2007-02-01
Budget End
2007-11-30
Support Year
2
Fiscal Year
2007
Total Cost
$202,479
Indirect Cost
Name
Thomas Jefferson University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
053284659
City
Philadelphia
State
PA
Country
United States
Zip Code
19107
Al-Saleem, Fetweh H; Sharma, Rashmi; Puligedda, Rama Devudu et al. (2017) RBC Adherence of Immune Complexes Containing Botulinum Toxin Improves Neutralization and Macrophage Uptake. Toxins (Basel) 9:
Sharma, Rashmi; Zhao, Huiwu; Al-Saleem, Fetweh H et al. (2014) Mechanisms of enhanced neutralization of botulinum neurotoxin by monoclonal antibodies conjugated to antibodies specific for the erythrocyte complement receptor. Mol Immunol 57:247-54
Benefield, Desirée A; Dessain, Scott K; Shine, Nancy et al. (2013) Molecular assembly of botulinum neurotoxin progenitor complexes. Proc Natl Acad Sci U S A 110:5630-5
Adekar, Sharad P; Segan, Andrew T; Chen, Cindy et al. (2011) Enhanced neutralization potency of botulinum neurotoxin antibodies using a red blood cell-targeting fusion protein. PLoS One 6:e17491
Adekar, Sharad P; Jones, R Mark; Elias, M D et al. (2008) Hybridoma populations enriched for affinity-matured human IgGs yield high-affinity antibodies specific for botulinum neurotoxins. J Immunol Methods 333:156-66
Adekar, Sharad P; Takahashi, Tsuyoshi; Jones, R Mark et al. (2008) Neutralization of botulinum neurotoxin by a human monoclonal antibody specific for the catalytic light chain. PLoS One 3:e3023
Adekar, Sharad P; Al-Saleem, Fetweh H; Elias, M D et al. (2008) A natural human IgM antibody that neutralizes botulinum neurotoxin in vivo. Hybridoma (Larchmt) 27:65-9
Dessain, S K; Adekar, S P; Berry, J D (2008) Exploring the native human antibody repertoire to create antiviral therapeutics. Curr Top Microbiol Immunol 317:155-83
Adekar, Sharad P; Jones, R Mark; Elias, M D et al. (2008) A human monoclonal antibody that binds serotype A botulinum neurotoxin. Hybridoma (Larchmt) 27:11-7