Vaccine induction of broadly neutralizing antibodies (bnAbs) to HIV remains a major challenge. Germline-targeting immunogens hold promise for initiating the induction of certain bnAb classes; yet for most bnAbs, a strong dependence on antibody heavy chain complementarity-determining region 3 (HCDR3) is a major barrier. Exploiting ultradeep human antibody sequencing data, we identified a diverse set of potential antibody precursors for a bnAb with dominant HCDR3 contacts. We then developed HIV envelope trimer–based immunogens that primed responses from rare bnAb-precursor B cells in a mouse model and bound a range of potential bnAb-precursor human naïve B cells in ex vivo screens. Our repertoire-guided germline-targeting approach provides a framework for priming the induction of many HIV bnAbs and could be applied to most HCDR3-dominant antibodies from other pathogens.
SUMMARYUnderstanding how broadly neutralizing antibodies (bnAbs) to HIV envelope
(Env) develop during natural infection can help guide the rational design of an
HIV vaccine. Here, we described a bnAb lineage targeting the Env V2 apex and the
Ab-Env co-evolution that led to development of neutralization breadth. The
lineage Abs bore an anionic heavy chain complementarity-determining region 3
(CDRH3) of 25 amino acids, among the shortest known for this class of Abs, and
achieved breadth with only 10% nucleotide somatic hypermutation and no
insertions or deletions. The data suggested a role for Env glycoform
heterogeneity in the activation of the lineage germ-line B cell. Finally, we
showed that localized diversity at key V2 epitope residues drove bnAb maturation
toward breadth, mirroring the Env evolution pattern described for another donor
who developed V2-apex targeting bnAbs. Overall, these findings suggest potential
strategies for vaccine approaches based on germline-targeting and serial
immunogen design.
Summary
The VH1-2 restricted VRC01-class of antibodies targeting the HIV envelope CD4 binding site are a major focus of HIV vaccine strategies. However, a detailed analysis of VRC01-class antibody development has been limited by the rare nature of these responses during natural infection and the lack of longitudinal sampling of such responses. To inform vaccine strategies, we mapped the development of a VRC01-class antibody lineage (PCIN63) in the subtype C infected IAVI Protocol C neutralizer PC063. PCIN63 monoclonal antibodies had the hallmark VRC01-class features and demonstrated neutralization breadth similar to the prototype VRC01 antibody, but were 2- to 3-fold less mutated. Maturation occurred rapidly within ∼24 months of emergence of the lineage and somatic hypermutations accumulated at key contact residues. This longitudinal study of broadly neutralizing VRC01-class antibody lineage reveals early binding to the N276-glycan during affinity maturation, which may have implications for vaccine design.
SummaryBroadly neutralizing antibodies (bnAbs) targeting the HIV envelope glycoprotein (Env) typically take years to develop. Longitudinal analyses of both neutralizing antibody lineages and viruses at serial time points during infection provide a basis for understanding the co-evolutionary contest between HIV and the humoral immune system. Here, we describe the structural characterization of an apex-targeting antibody lineage and autologous clade A viral Env from a donor in the Protocol C cohort. Comparison of Ab-Env complexes at early and late time points reveals that, within the antibody lineage, the CDRH3 loop rigidifies, the bnAb angle of approach steepens, and surface charges are mutated to accommodate glycan changes. Additionally, we observed differences in site-specific glycosylation between soluble and full-length Env constructs, which may be important for tuning optimal immunogenicity in soluble Env trimers. These studies therefore provide important guideposts for design of immunogens that prime and mature nAb responses to the Env V2-apex.
A nitrosopurine-ene reaction easily assembles the asmarine pharmacophore and transmits remote stereochemistry to the diazepine-purine hetereocycle. This reaction generates potent cytotoxins that exceed the potency of asmarine A (1.2 μM IC 50 ) and supersede the metabolites as useful leads for biological discovery.
Keywordsalkaloids; terpenoids; nitroso; purine; ene reaction Asmarines A and B (1&2, Figure 1) were identified in 1998 by Kashman and co-workers as the bioactive constituents of a Red Sea sponge (Raspailia sp.) extract, exhibiting cytotoxicity against several cancer cell lines with a minimum EC 50 of 1.2 μM and 120 nM, respectively. [1] The asmarines are unique among alkaloids by virtue of the embedded Nhydroxypurine diazepine (primary pharmacophore) [2] connected by an ethyl bridge to a
We describe a general
method to synthesize the iminium tetrahydrothiophene
embedded in the dimeric Nuphar alkaloids. In contrast
to prior studies, the sulfur atom of the thiaspirane pharmacophore
is shown to be electrophilic. This α-thioether reacts with thiophenol
or glutathione at ambient temperature to cleave the C–S bond
and form a disulfide. Rates of conversion are proportional to the
corresponding ammonium ion pKa and exhibit
half-lives less than 5 h at a 5 mM concentration of thiol. A simple
thiophane analogue of the Nuphar dimers causes apoptosis
at single-digit micromolar concentration and labels reactive cysteines
at similar levels as the unsaturated iminium “warhead”.
Our experiments combined with prior observations suggest the sulfur
of the Nuphar dimers can react as an electrophile
in cellular environments and that sulfur-triggered retrodimerization
can occur in the cell.
Wake up, protein! Small molecules that directly activate proteins are rare and their discovery opens new avenues for the development of drugs and chemical tools to probe the functions and mechanisms of protein targets. To address the one-sided dichotomy between enzyme inhibition and activation, we describe a series of procaspase activators as chemical tools in the study of caspase biology.
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