Passive immunotherapeutics are important in post exposure prophylaxis and treatment of infectious diseases. Reagents for passive immunotherapy include convalescent sera and monoclonal antibodies (mAb). Murine, humanized and fully human mAb (fhmAb) share advantages with respect to defined specificity, and enhanced safety profiles. However, only fhmAb possess native effector functions and eliminate risk of immune responses to nonhuman components that have the potential to reduce efficacy and/or cause autoimmune disease. FhmAb that neutralize Rift Valley fever virus (RVFV), Nipah/Hendra virus (NiV/HeV), and West Nile virus (WNV) were created using a fully human fusion partner cell line (MFP-2) and PEL from patients after natural infection (NiV and WNV) or vaccination (RVFV).
In Aim 1 of this project we will characterize these fhmAb in vivo. PBL from immune donors can difficult to obtain, particularly early in the course of an outbreak;thus, alternative strategies are needed whereby fhmAbs with optimized reactivity can be generated using synthetic reagents and in vitro maturation methods. Toward this end we will test (in Aims 2 and 3) the utility of a genetically engineered Veloclmmune? mouse for producing fhmAb from recombinant antigens representing NiV and Chikungunya virus (CHIKV). Functionality of Veloclmmune-drived mAb to NiV will be compared to that of the mAb generated with MFP-2. Furthermore, (Aim 4) we will apply two complementary methods for in vitro optimization of mAb reactivity: a yeast dual-display system that allows for in vitro mutation of cloned sequences followed by high throughput selection of reactive variants in a yeast system, and somatic hypermutation using the Ramos cell and/or activation induced cytidine deaminase (AID) systems.
Specific Aims are: (1) Characterize biological efficacy of existing fhmAb to RVFV, NiV/HeV and WNV;( 2) Generate fhmAb from protective Veloclmmune mAb to NiV and CHIKV;(3) Characterize Veloclmmune immune response and functionality of generated mAb;(4) Use recombinant genetics, mutagenesis, and somatic hypermutation to optimize mAb reactivity.

Public Health Relevance

Passive immunotherapy has an established track record in post exposure prophylaxis and treatment of infectious diseases and will continue to be a significant primary or complementary line of defense. With this project we will generate safe and specific fully human monoclonal antibody (fhmAb) therapeutics for Category A-C agents and establish a universal platform for efficiently generating these tools from entirely synthetic source materials, enabling a swift response to newly identified disease threats.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Specialized Center--Cooperative Agreements (U54)
Project #
5U54AI057158-07
Application #
8043570
Study Section
Special Emphasis Panel (ZAI1)
Project Start
Project End
Budget Start
2010-03-01
Budget End
2011-02-28
Support Year
7
Fiscal Year
2010
Total Cost
$515,568
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
Li, Xiao-Ping; Kahn, Jennifer N; Tumer, Nilgun E (2018) Peptide Mimics of the Ribosomal P Stalk Inhibit the Activity of Ricin A Chain by Preventing Ribosome Binding. Toxins (Basel) 10:
Goldman, David L; Nieves, Edward; Nakouzi, Antonio et al. (2018) Serum-Mediated Cleavage of Bacillus anthracis Protective Antigen Is a Two-Step Process That Involves a Serum Carboxypeptidase. mSphere 3:
Marié, Isabelle J; Chang, Hao-Ming; Levy, David E (2018) HDAC stimulates gene expression through BRD4 availability in response to IFN and in interferonopathies. J Exp Med 215:3194-3212
Uhde, Melanie; Ajamian, Mary; Wormser, Gary P et al. (2017) Reply to Naktin. Clin Infect Dis 64:1145-1146
Chen, Han; Coseno, Molly; Ficarro, Scott B et al. (2017) A Small Covalent Allosteric Inhibitor of Human Cytomegalovirus DNA Polymerase Subunit Interactions. ACS Infect Dis 3:112-118
Aguilar, Jorge L; Varshney, Avanish K; Pechuan, Ximo et al. (2017) Monoclonal antibodies protect from Staphylococcal Enterotoxin K (SEK) induced toxic shock and sepsis by USA300 Staphylococcus aureus. Virulence 8:741-750
Zhou, Yijun; Li, Xiao-Ping; Chen, Brian Y et al. (2017) Ricin uses arginine 235 as an anchor residue to bind to P-proteins of the ribosomal stalk. Sci Rep 7:42912
Lauretti, Flavio; Chattopadhyay, Anasuya; de Oliveira França, Rafael Freitas et al. (2016) Recombinant vesicular stomatitis virus-based dengue-2 vaccine candidate induces humoral response and protects mice against lethal infection. Hum Vaccin Immunother 12:2327-33
Tadin, Ante; Tokarz, Rafal; Markoti?, Alemka et al. (2016) Molecular Survey of Zoonotic Agents in Rodents and Other Small Mammals in Croatia. Am J Trop Med Hyg 94:466-73
Basu, Debaleena; Li, Xiao-Ping; Kahn, Jennifer N et al. (2016) The A1 Subunit of Shiga Toxin 2 Has Higher Affinity for Ribosomes and Higher Catalytic Activity than the A1 Subunit of Shiga Toxin 1. Infect Immun 84:149-61

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