Engineering an Antibody V Gene-Selective Vaccine

Ronsard, Larance; Yousif, Ashraf S.; Peabody, Julianne; Okonkwo, Vintus; Devant, Pascal; Mogus, Alemu Tekewe; Barnes, Ralston M.; Rohrer, Daniel K.; Lönberg, Nils; Peabody, David S.; Chackerian, Bryce; Lingwood, Daniel

doi:10.3389/fimmu.2021.730471

Cited by 6 publications

(7 citation statements)

References 94 publications

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“…Despite this 4-fold difference in affinity, we observed comparable total BCR signaling relative to the IgM control for all functionalized DNA-VLPs, consistent with previously described avidity effects at the B-cell surface 61 . We concluded that our DNA-VLPs efficiently interacted with and induced signaling by RBD-specific BCRs, analogous to previous studies using similar assays to evaluate multivalent subunit vaccines 58,[62][63][64][65][66][67][68] . The increased B-cell activation for I52-30x-RBD contrasts our previous findings for HIV antigens for which total Ca 2+ flux saturated beyond a valency of 10x 48 .…”

Section: Resultssupporting

confidence: 83%

Enhancing antibody responses by multivalent antigen display on thymus-independent DNA origami scaffolds

Wamhoff

Ronsard

Feldman

et al. 2022

Preprint

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Multivalent antigen display is a well-established design principle to enhance humoral immunity elicited by subunit vaccines. Protein-based virus-like particles (VLPs) are an important vaccine platform that implements this principle but also contain thymus-dependent off-target epitopes, thereby generating neutralizing and defocused antibody responses against the scaffold itself. Here, we present DNA origami as an alternative platform to display the receptor binding domain (RBD) of SARS-CoV-2. DNA-based scaffolds provide nanoscale control over antigen organization and, as thymus-independent antigens, are expected to induce only extrafollicular B-cell responses. Our icosahedral DNA-based VLPs elicited valency-dependent BCR signaling in two reporter B-cell lines, with corresponding increases in RBD-specific antibody responses following sequential immunization in mice. Mouse sera also neutralized the Wuhan strain of SARS-CoV-2 - but did not contain boosted, DNA-specific antibodies. Thus, multivalent display using DNA origami can enhance immunogenicity of protein antigens without generating scaffold-directed immunological memory and may prove useful for rational vaccine design.

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Section: Resultssupporting

confidence: 83%

Enhancing antibody responses by multivalent antigen display on thymus-independent DNA origami scaffolds

Wamhoff

Ronsard

Feldman

et al. 2022

Preprint

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show abstract

“…Despite this four-fold difference in affinity, we observed comparable total BCR signaling relative to the IgM control for all functionalized DNA-VLPs, consistent with previously described avidity effects at the B cell surface 63 . We conclude that our DNA-VLPs efficiently bound and induced signaling by RBD-specific BCRs in a valency-dependent manner, analogous to studies using similar assays to evaluate protein-and DNA-scaffolded multivalent subunit vaccines 53,61,[64][65][66][67][68][69][70] .…”

Section: Dna-vlps Elicit Valency-dependent B Cell Signaling In Vitrosupporting

confidence: 53%

“…The B cell activation assay was adapted from previously established protocols 61 , 69 , 98 . Briefly, the human anti-RBD antibodies CR3022 59 and B38 62 were expressed as IgM B cell receptors (BCRs) in Ramos B cells after lentiviral transfection of the corresponding light chain and transmembrane IgM heavy chain genes.…”

Section: Methodsmentioning

confidence: 99%

Enhancing antibody responses by multivalent antigen display on thymus-independent DNA origami scaffolds

Wamhoff,

Ronsard,

Feldman

et al. 2024

Nat Commun

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Protein-based virus-like particles (P-VLPs) are commonly used to spatially organize antigens and enhance humoral immunity through multivalent antigen display. However, P-VLPs are thymus-dependent antigens that are themselves immunogenic and can induce B cell responses that may neutralize the platform. Here, we investigate thymus-independent DNA origami as an alternative material for multivalent antigen display using the receptor binding domain (RBD) of the SARS-CoV-2 spike protein, the primary target of neutralizing antibody responses. Sequential immunization of mice with DNA-based VLPs (DNA-VLPs) elicits protective neutralizing antibodies to SARS-CoV-2 in a manner that depends on the valency of the antigen displayed and on T cell help. Importantly, the immune sera do not contain boosted, class-switched antibodies against the DNA scaffold, in contrast to P-VLPs that elicit strong B cell memory against both the target antigen and the scaffold. Thus, DNA-VLPs enhance target antigen immunogenicity without generating scaffold-directed immunity and thereby offer an important alternative material for particulate vaccine design.

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“…The HC2 mice are an established transgenic model in which human V H gene usage is constrained to user-defined gene segments, while allowing for recombination with diverse human D and J segments, generating an antibody CDRH3 repertoire that is similar to humans and which supports affinity matured antibodies following immunization with protein antigens 47 – 52 . HC2 mice using the human V H gene IGHV1-2*02 were deployed in which LC input was from the mouse repertoire 47 , 50 – 52 . The HC2 mice were a gift to D.L.…”

Section: Methodsmentioning

confidence: 99%

Engaging an HIV vaccine target through the acquisition of low B cell affinity

Ronsard,

Yousif,

Nait Mohamed

et al. 2023

Nat Commun

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Low affinity is common for germline B cell receptors (BCR) seeding development of broadly neutralizing antibodies (bnAbs) that engage hypervariable viruses, including HIV. Antibody affinity selection is also non-homogenizing, insuring the survival of low affinity B cell clones. To explore whether this provides a natural window for expanding human B cell lineages against conserved vaccine targets, we deploy transgenic mice mimicking human antibody diversity and somatic hypermutation (SHM) and immunize with simple monomeric HIV glycoprotein envelope immunogens. We report an immunization regimen that focuses B cell memory upon the conserved CD4 binding site (CD4bs) through both conventional affinity maturation and reproducible expansion of low affinity BCR clones with public patterns in SHM. In the latter instance, SHM facilitates target acquisition by decreasing binding strength. This suggests that permissive B cell selection enables the discovery of antibody epitopes, in this case an HIV bnAb site.

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Engineering an Antibody V Gene-Selective Vaccine

Cited by 6 publications

References 94 publications

Enhancing antibody responses by multivalent antigen display on thymus-independent DNA origami scaffolds

Enhancing antibody responses by multivalent antigen display on thymus-independent DNA origami scaffolds

Enhancing antibody responses by multivalent antigen display on thymus-independent DNA origami scaffolds

Engaging an HIV vaccine target through the acquisition of low B cell affinity

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