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TitleElectron-Microscopy-Based Epitope Mapping Defines Specificities of Polyclonal Antibodies Elicited during HIV-1 BG505 Envelope Trimer Immunization.
Journal, issue, pagesImmunity, Vol. 49, Issue 2, Page 288-300.e8, Year 2018
Publish dateAug 21, 2018
AuthorsMatteo Bianchi / Hannah L Turner / Bartek Nogal / Christopher A Cottrell / David Oyen / Matthias Pauthner / Raiza Bastidas / Rebecca Nedellec / Laura E McCoy / Ian A Wilson / Dennis R Burton / Andrew B Ward / Lars Hangartner /
PubMed AbstractCharacterizing polyclonal antibody responses via currently available methods is inherently complex and difficult. Mapping epitopes in an immune response is typically incomplete, which creates a ...Characterizing polyclonal antibody responses via currently available methods is inherently complex and difficult. Mapping epitopes in an immune response is typically incomplete, which creates a barrier to fully understanding the humoral response to antigens and hinders rational vaccine design efforts. Here, we describe a method of characterizing polyclonal responses by using electron microscopy, and we applied this method to the immunization of rabbits with an HIV-1 envelope glycoprotein vaccine candidate, BG505 SOSIP.664. We detected known epitopes within the polyclonal sera and revealed how antibody responses evolved during the prime-boosting strategy to ultimately result in a neutralizing antibody response. We uncovered previously unidentified epitopes, including an epitope proximal to one recognized by human broadly neutralizing antibodies as well as potentially distracting non-neutralizing epitopes. Our method provides an efficient and semiquantitative map of epitopes that are targeted in a polyclonal antibody response and should be of widespread utility in vaccine and infection studies.
External linksPubMed:30097292 / Publisher's page
KeywordsAIDS Vaccines / Animals / Antibodies, Neutralizing / Antibody Formation / Cell Line / Epitope Mapping / Epitopes / Female / HEK293 Cells / HIV Antibodies / HIV Infections / HIV-1 / Humans / Immunization / Immunoglobulin Fab Fragments / Microscopy, Electron / Rabbits / Recombinant Proteins / env Gene Products, Human Immunodeficiency Virus / IMMUNE SYSTEM / Fab / HIV / VIRAL PROTEIN / antibodies / antibody
MethodsEM (single particle) / X-ray diffraction
Resolution1.795 - 20 A
Structure data

EMDB-7552:
CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417

EMDB-7553:
CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417

EMDB-7554:
CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417

EMDB-7555:
CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417

EMDB-7556:
CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417

EMDB-7557:
CryoEM map of BG505 SOSIP.664 in complex with post boost 1 serum from rabbit 3417

EMDB-7570:
Negative stain EM map of BG505 SOSIP.644 in complex with PG9 and 12N Fabs

EMDB-7887:
Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3418 at post boost 2

EMDB-7888:
Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3418 at post boost 2.

EMDB-7889:
Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3419 at post boost 2.

EMDB-7890:
Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3419 at post boost 2.

EMDB-7891:
Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3417 at post boost 2.

EMDB-7892:
Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3417 at post boost 2.

EMDB-7893:
Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3417 at post boost 2.

EMDB-7894:
Negative Stain EM map of polyclonal serum in complex with BG505 SOSIP.664 from rabbit 3420 at post boost 2.

EMDB-7895:
Negative Stain EM map of polyclonal serum in complex with BG505 MD39 CPG9 from rabbit 3417 at post boost 3

EMDB-7896:
HIV Env BG505 SOSIP with polyclonal Fabs from immunized rabbit #3417 post-boost#1

EMDB-7903:
Negative stain EM map of BG505 SOSIP.644 in complex with PGT121 and 12N Fabs

EMDB-7904:
Negative stain EM map of BG505 SOSIP.644 in complex with PGT121 and serum from rabbit 3417 PB1

EMDB-7906:
Negative stain EM map of BG505 SOSIP.644 in complex with PG9 Fab and serum from rabbit 3417 PB1

PDB-6cjk:
Anti HIV Fab 10A

PDB-6did:
HIV Env BG505 SOSIP with polyclonal Fabs from immunized rabbit #3417 post-boost#1

Chemicals

ChemComp-ACT:
ACETATE ION / Acetate

ChemComp-GOL:
GLYCEROL / Glycerol

ChemComp-HOH:
WATER / Water

ChemComp-NAG:
N-ACETYL-D-GLUCOSAMINE / N-Acetylglucosamine

ChemComp-BMA:
BETA-D-MANNOSE / Mannose

ChemComp-MAN:
ALPHA-D-MANNOSE / Mannose

Source
  • Human immunodeficiency virus
  • oryctolagus cuniculus (rabbit)
  • Homo sapiens (human)
  • human immunodeficiency virus 1

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