1 / 16

Jacob Glanville

Neutralizing the VSG Repertoire. Jacob Glanville. Trypanosoma brucei and the Variable Surface Glycoprotein (VSG). Anti-VSG antibodies neutralize T Brucei.

aneko
Download Presentation

Jacob Glanville

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Neutralizing the VSG Repertoire Jacob Glanville

  2. Trypanosomabrucei and the Variable Surface Glycoprotein (VSG)

  3. Anti-VSG antibodies neutralize T Brucei Stijlemans B, Caljon G, Natesan SKA, Saerens D, et al. (2011) High Affinity Nanobodies against the Trypanosome brucei VSG Are Potent Trypanolytic Agents that Block Endocytosis. PLoSPathog 7(6): e1002072. doi:10.1371/journal.ppat.1002072

  4. Immunodominance & cryptic epitopes

  5. If bnMabs can be made, why don’t we all make them? If in HA and HIV, why not VSG?

  6. Hypothesis: There exist conserved epitopes in the VSG repertoire that cannot escape antibody recognition.

  7. Aim 1: Identify conserved candidate broadly neutralizing epitopes from high-throughput primary and tertiary structure analysis of the VSG repertoire • Sequence many people • Map variation to structure • Identify putative conserved epitopes

  8. Trypanosomes from 100-1000 patients • 5’RACE cDNAamplification • SafeSeqS protocol • Multiplex barcoding (5-50 samples/run) • Miseq (>5M 250x2 paired-end reads) • Profile Hidden Markov Models • Structural modeling • Estimating recovery • 10 samples pooled per run such that each patient sample is sequenced to an approximate depth of 500,000 250x2 paired end reads. Given an assumed mean minor VSG frequency of 1e-4 and dominant VSG at 99%, we can expect to recover ~100 unique active VSGs per patient, and ~1,000 active VSGs per sequencing run, at 50x coverage. Given 200 patients over 20 sequencing runs, a database of 20,000 VSGs could be recovered for analysis.

  9. Aim 2: Recover cross-reactive anti-VSG antibodies from human patients. • Generate protein panel of VSGs • Test serum of many infected people • Single cell isolate B-cells and test monoclonal VDG reactivity profiles • Epitope mapping

  10. 1800 patients -> 1 primary candidate -> 30,000 single cells -> 2 bnAbs

  11. Significance Database for analyzing VSG Diversity dynamics Reagents for analyzing VSG antigens Practical outcome for vaccine design Fundamental implications for immune evasion

  12. Take-away Pathogens use immunodominance and antigenic drift to rig the B-cell activation race so that the losers always win.

More Related