This proposal focuses on identifying and diversifying novel monoclonal antibodies (Mabs) isolated from humans (huMabs) that are exposed to viral sexually transmitted pathogens. Coupling antibody discovery technology (FACS/RT-PCR) with a versatile and rapid (days to weeks) production system facilitates evaluation of the greatest range of protective huMabs. The variable regions isolated from STI exposed individuals when matched with an array of pre-built scaffolds that have defined constant region features (e.g. multimeric, + complement binding), significantly increases the diversity and utility of the STI Mab portfolio for understanding antibody correlates of protection for vaccines and microbicides.In the first Specific Aim, we will isolate and clone huMabs from STI exposed individuals. Initial efforts will focus on isolation of human Mabs from the antibody repertoire of HIV exposed uninfected (HIV-EU) individuals. In the later years of the project, efforts will be expanded to HSV-2 and HPV seropositive individuals, as well as individuals immunized in Project 3 with the experimental vaccines generated by the Production Core. Antigen-specific IgG+, IgM+, and IgA+ B cells from these individuals will be subjected to single ceil sorting (FACS) against antigens identified as being relevant in the immunity observed against the specific pathogen. The antibody variable genes will be cloned by RT-PCR, and inserted in pre-built vector scaffolds of constant regions with defined features (e.g. non-inflammatory, stable in mucus, valency, potential to perform effector functions). These huMabs will then be expressed as part of the ProductionCore in a transient plant system capable of producing milligram quantities in 1-2 weeks for in vitro and in vivo evaluation.In the second Specific Aim, huMabs will be evaluated in vitro for neutralization activity and somehuMabs will be further characterized in mouse vaginal challenge models that have been established for evaluating microbicides and vaccines. Resulting data will help identify antibody correlates of protection from STIs (e.g. identification of protective epitopes, the role of antibody isotype, and the neutralizing titer of mucosal antibodies necessary for protection), providing guidance in vaccine development, and may yield huMabs worthy of clinical development as microbicides.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program--Cooperative Agreements (U19)
Project #
1U19AI062150-01
Application #
6866240
Study Section
Special Emphasis Panel (ZAI1-NBS-M (M4))
Project Start
2004-08-01
Project End
2009-07-31
Budget Start
2004-09-01
Budget End
2005-08-31
Support Year
1
Fiscal Year
2004
Total Cost
$308,621
Indirect Cost
Name
Arizona State University-Tempe Campus
Department
Type
DUNS #
943360412
City
Tempe
State
AZ
Country
United States
Zip Code
85287
Meador, Lydia R; Kessans, Sarah A; Kilbourne, Jacquelyn et al. (2017) A heterologous prime-boosting strategy with replicating Vaccinia virus vectors and plant-produced HIV-1 Gag/dgp41 virus-like particles. Virology 507:242-256
Kessans, Sarah A; Linhart, Mark D; Meador, Lydia R et al. (2016) Immunological Characterization of Plant-Based HIV-1 Gag/Dgp41 Virus-Like Particles. PLoS One 11:e0151842
Mathew, Lolita George; Herbst-Kralovetz, Melissa M; Mason, Hugh S (2014) Norovirus Narita 104 virus-like particles expressed in Nicotiana benthamiana induce serum and mucosal immune responses. Biomed Res Int 2014:807539
Whaley, Kevin J; Morton, Josh; Hume, Steve et al. (2014) Emerging antibody-based products. Curr Top Microbiol Immunol 375:107-26
Lee, Ho-Hsien; Cherni, Irene; Yu, HongQi et al. (2014) Expression, purification and crystallization of CTB-MPR, a candidate mucosal vaccine component against HIV-1. IUCrJ 1:305-17
Hjelm, Brooke E; Kilbourne, Jacquelyn; Herbst-Kralovetz, Melissa M (2014) TLR7 and 9 agonists are highly effective mucosal adjuvants for norovirus virus-like particle vaccines. Hum Vaccin Immunother 10:410-6
McGowin, Chris L; Radtke, Andrea L; Abraham, Kyle et al. (2013) Mycoplasma genitalium infection activates cellular host defense and inflammation pathways in a 3-dimensional human endocervical epithelial cell model. J Infect Dis 207:1857-68
Kessans, Sarah A; Linhart, Mark D; Matoba, Nobuyuki et al. (2013) Biological and biochemical characterization of HIV-1 Gag/dgp41 virus-like particles expressed in Nicotiana benthamiana. Plant Biotechnol J 11:681-90
Lai, Huafang; Chen, Qiang (2012) Bioprocessing of plant-derived virus-like particles of Norwalk virus capsid protein under current Good Manufacture Practice regulations. Plant Cell Rep 31:573-84
Jackson, Erin M; Herbst-Kralovetz, Melissa M (2012) Intranasal vaccination with murabutide enhances humoral and mucosal immune responses to a virus-like particle vaccine. PLoS One 7:e41529

Showing the most recent 10 out of 32 publications