A5005979779- HIV Research and Treatment
- Monoclonal and Polyclonal Antibodies Research
- Immune Cell Function and Interaction
- HIV/AIDS drug development and treatment
- vaccines and immunoinformatics approaches
- Glycosylation and Glycoproteins Research
- Blood groups and transfusion
- Influenza Virus Research Studies
- Enzyme Structure and Function
- Hepatitis C virus research
- T-cell and B-cell Immunology
- Viral Infections and Outbreaks Research
- Protein purification and stability
- Hepatitis B Virus Studies
- Bacteriophages and microbial interactions
- RNA Interference and Gene Delivery
- Immunotherapy and Immune Responses
- Systemic Lupus Erythematosus Research
- Chronic Lymphocytic Leukemia Research
- Viral Infections and Vectors
- Virus-based gene therapy research
- Immunodeficiency and Autoimmune Disorders
- Transgenic Plants and Applications
- Poxvirus research and outbreaks
- thermodynamics and calorimetric analyses
University of Maryland, Baltimore
2014-2024
National Institute of Standards and Technology
2014-2024
University of Maryland, College Park
2014-2024
Institute for Bioscience and Biotechnology Research
2014-2024
Advanced Bioscience Laboratories (United States)
2022-2024
National Institute of Allergy and Infectious Diseases
2004-2016
National Institutes of Health
2003-2016
Simon Fraser University
2013
National Cancer Institute
2013
Institut de Recherche Vaccinale
2012-2013
The membrane-proximal region of the ectodomain gp41 envelope glycoprotein human immunodeficiency virus type 1 (HIV-1) is target three five broadly neutralizing anti-HIV-1 antibodies thus far isolated. We have determined crystal structures antigen-binding fragment for one these antibodies, 2F5, in complex with 7-mer, 11-mer, and 17-mer peptides region, at 2.0-, 2.1-, 2.2-A resolutions, respectively. reveal an extended conformation, which stretches over 30 A length. Contacts are made...
Elicitation of antibodies against targets that are immunorecessive, cryptic, or transient in their native context has been a challenge for vaccine design. Here we demonstrate the elicitation structure-specific HIV-1 gp41 epitope broadly neutralizing antibody 2F5. This conformationally flexible region assumes mostly helical conformations but adopts kinked, extended structure when bound by Computational techniques were employed to transplant 2F5 into select acceptor scaffolds. The resultant...
Serum characterization and antibody isolation are transforming our understanding of the humoral immune response to viral infection. Here, we show that epitope specificities HIV-1-neutralizing antibodies in serum can be elucidated from pattern neutralization against a diverse panel HIV-1 isolates. We determined "neutralization fingerprints" for 30 neutralizing on 34 strains showed similarity fingerprint correlated with epitope. used these fingerprints delineate polyclonal sera 24...
A critical roadblock to HIV vaccine development is the inability induce B cell lineages of broadly neutralizing antibodies (bnAbs) in humans. In people living with HIV-1, bnAbs take years develop. The HVTN 133 clinical trial studied a peptide/liposome immunogen targeting HIV-1 envelope (Env) membrane-proximal external region (MPER) (NCT03934541). Here, we report MPER peptide-liposome induction polyclonal mature and their precursors, most potent which neutralized 15% global tier 2 strains 35%...
Next-generation sequencing of antibody transcripts from HIV-1-infected individuals with broadly neutralizing antibodies could provide an efficient means for identifying somatic variants and characterizing their lineages. Here, we used 454 pyrosequencing identity/divergence grid sampling to analyze heavy- light-chain sequences donor N152, the source 10E8. We identified up 28% difference in amino acid sequence. Heavy- phylogenetic trees 10E8 displayed similar architectures, reconstituted...
Induction of broadly neutralizing antibodies (bnAbs) is a goal HIV-1 vaccine development. Antibody 10E8, reactive with the distal portion membrane-proximal external region (MPER) gp41, neutralizing. However, ontogeny MPER and relationship memory B cell to plasma bnAbs are poorly understood. HIV-1-specific flow sorting proteomic identification anti-MPER from an HIV-1-infected individual were used isolate same clonal lineage. Structural analysis demonstrated that cells recognized envelope gp41...
The membrane-proximal external region (MPER) of the HIV-1 gp41 transmembrane glycoprotein is target broadly neutralizing antibody 2F5. Prior studies have suggested a two-component mechanism for 2F5-mediated neutralization involving both structure-specific recognition protein epitope and nonspecific interaction with viral lipid membrane. Here, we mutationally alter hydrophobic patch on third complementarity-determining heavy chain (CDR H3) 2F5 assess abilities altered variants to bind...
Abs capable of effectively neutralizing HIV-1 generally exhibit very high levels somatic hypermutation, both in their CDR and framework-variable regions. In many cases, full reversion the Ab-framework mutations back to germline results substantial complete loss HIV-1-neutralizing activity. However, it has been unclear whether all or most observed framework would be necessary a small subset these might sufficient for broad potent neutralization. To address this issue explore dependence...
The genes encoding broadly HIV-1-neutralizing human monoclonal antibodies (MAbs) are highly divergent from their germ line counterparts. We have hypothesized that such high levels of somatic hypermutation could pose a challenge for elicitation the neutralizing (bn) Abs and identification less somatically mutated bn may help in design effective vaccine immunogens. In quest Abs, phage- yeast-displayed antibody libraries, constructed using peripheral blood mononuclear cells (PBMCs) patient with...
The HIV-1 envelope glycoproteins (Env) gp120 and gp41 mediate entry are the targets for neutralizing antibodies. Within gp41, a continuous epitope defined by broadly antibody 2F5, is one of few conserved sites accessible to antibodies on functional HIV Env spike. Recently, as an initial attempt at structure-guided design, we transplanted 2F5 onto several non-HIV acceptor scaffold proteins that termed scaffolds (ES). As immunogens, these ES elicited with exquisite binding specificity matching...
ABSTRACT Extraordinary antibodies capable of near pan-neutralization HIV-1 have been identified. One the broadest is antibody 10E8, which recognizes membrane-proximal external region (MPER) envelope and neutralizes >95% circulating strains. If delivered passively, 10E8 might serve to prevent or treat infection. Antibody however, markedly less soluble than other antibodies. Here, we describe use both structural biology somatic variation develop optimized versions with increased solubility....
A component to the problem of inducing broad neutralizing HIV-1 gp41 membrane proximal external region (MPER) antibodies is need focus antibody response transiently exposed MPER pre-hairpin intermediate neutralization epitope. Here we describe a envelope (Env) gp140 oligomer prime followed by peptide-liposomes boost strategy for eliciting serum responses in rhesus macaques that bind fusion protein. This Env-liposome immunization induced 2F5 epitope 664DKW residues, and these preferentially...
One strategy for isolating or eliciting antibodies against a specific target region on the envelope glycoprotein trimer (Env) of human immunodeficiency virus type 1 (HIV-1) involves creation site transplants, which present heterologous protein scaffold with preserved antibody-binding properties. If is supersite HIV-1 vulnerability, recognized by collection broadly neutralizing antibodies, this affords "supersite transplants", capable binding (and potentially eliciting) similar to template...
Antibody 10E8 targets the membrane-proximal external region (MPER) of HIV-1 gp41, neutralizes >97% isolates, and lacks auto-reactivity often associated with MPER-directed antibodies. The developmental pathway might therefore serve as a promising template for vaccine design, but samples from time-of-infection-often used to infer B cell record-are unavailable. In this study, we crystallography, next-generation sequencing (NGS), functional assessments single time point. Mutational analysis...
Antibodies m66.6 and 2F5 are the only effective human HIV-1-neutralizing antibodies reported thus far to recognize N-terminal region of membrane-proximal external (MPER) gp41 subunit HIV-1 envelope glycoprotein. Although has been extensively characterized, much less is known about antibody or m66, a closely related light-chain variant. Here, we report crystal structure m66 in complex with its epitope, along unbound structures m66.6. We used mutational binding analyses decipher elements...
Abstract Hepatitis C virus (HCV) is a major global health burden as the leading causative agent of chronic liver disease and hepatocellular carcinoma. While main antigenic target for HCV-neutralizing antibodies membrane-associated E1E2 surface glycoprotein, development effective vaccines has been hindered by complications in biochemical preparation soluble ectodomains. Here, we present cryo-EM structure an engineered, secreted ectodomain genotype 1b complex with neutralizing AR4A, HEPC74,...
Hepatitis C virus (HCV) is a major medical health burden and the leading cause of chronic liver disease cancer worldwide. More than 58 million people are chronically infected with HCV, 1.5 new infections occurring each year. An effective HCV vaccine public need as recognized by World Health Organization. However, due to high variability its ability escape immune response, rapidly accumulates mutations, making development formidable challenge. must elicit broadly neutralizing antibodies...
The severity of the 2014-2016 ebolavirus outbreak in West Africa expedited clinical development therapeutics and vaccines though countermeasures on hand were largely monospecific lacked efficacy against other species that previously emerged. Recent studies indicate glycoprotein (GP) fusion loops are targets for cross-protective antibodies. Here we report 3.72 Å resolution crystal structure one such antibody, CA45, bound to ectodomain Ebola virus (EBOV) GP. CA45 epitope spans multiple faces...