Camelid VHH Antibodies that Neutralize Botulinum Neurotoxin Serotype E Intoxication or Protease Function

被引:18
作者
Tremblay, Jacqueline M. [1 ]
Vazquez-Cintron, Edwin [2 ]
Lam, Kwok-Ho [3 ]
Mukherjee, Jean [1 ]
Bedenice, Daniela [4 ]
Ondeck, Celinia A. [2 ]
Conroy, Matthieu T. [2 ]
Bodt, Skylar M. L. [2 ]
Winner, Brittany M. [2 ]
Webb, Robert P. [5 ]
Ichtchenko, Konstantin [6 ]
Jin, Rongsheng [3 ]
McNutt, Patrick M. [2 ]
Shoemaker, Charles B. [1 ]
机构
[1] Tufts Univ, Cummings Sch Vet Med, Dept Infect Dis & Global Hlth, North Grafton, MA 01536 USA
[2] US Army, Med Res Inst Chem Def, Aberdeen Proving Ground, MD 21010 USA
[3] Univ Calif Irvine, Dept Physiol & Biophys, Irvine, CA 92697 USA
[4] Tufts Univ, Cummings Sch Vet Med, Dept Clin Sci, North Grafton, MA 01536 USA
[5] US Army, Med Res Inst Infect Dis, Bacteriol Div, Ft Detrick, MD 21702 USA
[6] NYU, Sch Med, Dept Biochem & Mol Pharmacol, New York, NY 10016 USA
关键词
botulinum neurotoxin; botulism; toxin; antitoxin; single-domain antibody; VHH; neutralization; protease; SINGLE; BINDING; NANOBODIES; AGENTS; CHALLENGE; PROTECTS; DOMAINS; TOXINS; MICE; PH;
D O I
10.3390/toxins12100611
中图分类号
TS2 [食品工业];
学科分类号
0832 ;
摘要
Botulinum neurotoxin (BoNT) serotype E is one of three serotypes that cause the preponderance of human botulism cases and is a Tier 1 Select Agent. BoNT/E is unusual among BoNT serotypes for its rapid onset and short duration of intoxication. Here we report two large panels of unique, unrelated camelid single-domain antibodies (VHHs) that were selected for their ability to bind to BoNT/E holotoxin and/or to the BoNT/E light chain protease domain (LC/E). The 19 VHHs which bind to BoNT/E were characterized for their subunit specificity and 8 VHHs displayed the ability to neutralize BoNT/E intoxication of neurons. Heterodimer antitoxins consisting of two BoNT/E-neutralizing VHHs, including one heterodimer designed using structural information for simultaneous binding, were shown to protect mice against co-administered toxin challenges of up to 500 MIPLD50. The 22 unique VHHs which bind to LC/E were characterized for their binding properties and 9 displayed the ability to inhibit LC/E protease activity. Surprisingly, VHHs selected on plastic-coated LC/E were virtually unable to recognize soluble or captured LC/E while VHHs selected on captured LC/E were poorly able to recognize LC/E coated to a plastic surface. This panel of anti-LC/E VHHs offer insight into BoNT/E function, and some may have value as components of therapeutic antidotes that reverse paralysis following BoNT/E exposures.
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页数:18
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共 49 条
[1]   Nanobodies as novel therapeutic agents in envenomation [J].
Alirahimi, Ehsan ;
Kazemi-Lomedasht, Fatemeh ;
Shahbazzadeh, Delavar ;
Habibi-Anbouhi, Mandi ;
Chafi, Mohammad Hosseininejad ;
Sotoudeh, Nazli ;
Ghaderi, Hajarossadat ;
Muyldermans, Serge ;
Behdani, Mandi .
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 2018, 1862 (12) :2955-2965
[2]  
Anonymous, 2010, Morbidity and Mortality Weekly Report, V59, P299
[3]  
[Anonymous], 2011, GUIDE CARE USE LAB A, V8th
[4]   Creation and development of the public service orphan drug human botulism immune globulin [J].
Arnon, Stephen S. .
PEDIATRICS, 2007, 119 (04) :785-789
[5]   Botulinum neurotoxin type D enables cytosolic delivery of enzymatically active cargo proteins to neurones via unfolded translocation intermediates [J].
Bade, S ;
Rummel, A ;
Reisinger, C ;
Karnath, T ;
Ahnert-Hilger, G ;
Bigalke, H ;
Binz, T .
JOURNAL OF NEUROCHEMISTRY, 2004, 91 (06) :1461-1472
[6]   In Vivo Neutralization of Botulinum Neurotoxins Serotype E with Heavy-chain Camelid Antibodies (VHH) [J].
Bakherad, Hamid ;
Gargari, Seyed Latif Mousavi ;
Rasooli, Iraj ;
RajabiBazl, Masoumeh ;
Mohammadi, Mohammad ;
Ebrahimizadeh, Walead ;
Ardakani, Leila Safaee ;
Zare, Hamed .
MOLECULAR BIOTECHNOLOGY, 2013, 55 (02) :159-167
[7]   Botulinum and Tetanus Neurotoxin-Induced Blockade of Synaptic Transmission in Networked Cultures of Human and Rodent Neurons [J].
Beske, Phillip H. ;
Bradford, Aaron B. ;
Grynovicki, Justin O. ;
Glotfelty, Elliot J. ;
Hoffman, Katie M. ;
Hubbard, Kyle S. ;
Tuznik, Kaylie M. ;
McNutt, Patrick M. .
TOXICOLOGICAL SCIENCES, 2016, 149 (02) :503-515
[8]   Botulinum neurotoxin light chain refolds at endosomal pH for its translocation [J].
Cai, Shuowei ;
Kukreja, Roshan ;
Shoesmith, Sue ;
Chang, Tzuu-Wang ;
Singh, Bal Ram .
PROTEIN JOURNAL, 2006, 25 (7-8) :455-462
[9]   Heat-Labile Enterotoxin IIa, a Platform To Deliver Heterologous Proteins into Neurons [J].
Chen, Chen ;
Przedpelski, Amanda ;
Tepp, William H. ;
Pellett, Sabine ;
Johnson, Eric A. ;
Barbieri, Joseph T. .
MBIO, 2015, 6 (04)
[10]   Solubility of the catalytic domains of Botulinum neurotoxin serotype E subtypes [J].
Chen, Sheng ;
Barbieri, Joseph T. .
PROTEIN EXPRESSION AND PURIFICATION, 2016, 118 :18-24