Membrane-bound mRNA immunogens lower the threshold to activate HIV Env V2 apex-directed broadly neutralizing B cell precursors in humanized mice

Melzi, Eleonora; Willis, Jordan R.; Krystal, Mark; Lin, Ying‐Cing; Berndsen, Zachary T.; Landais, Elise; Oleksandr, Kalyuzhniy; Nair, Usha; Warner, John E.; Steichen, Jon M.; Kalyuzhniy, Anton; Le, Amber; Pecetta, Simone; Perez, Manfredo; Kirsch, Kathrin H.; Weldon, Stephanie R.; Falcone, Samantha; Himansu, Sunny; Carfı́, Andrea; Sok, Devin; Ward, Andrew B.; Schief, William R.; Batista, Facundo D.

doi:10.1016/j.immuni.2022.09.003

Cited by 28 publications

(24 citation statements)

References 90 publications

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“…We observed that a 1 μg dose of SOSIP-TM delivered as mRNA-LNP raised comparable neutralizing responses to 25 μg of SOSIP mi3-multimers, whereas 5 μg of SOSIP-TM mRNA-LNP elicited a significantly more potent neutralizing response. Note that 25 μg of adjuvanted protein antigen is a high dose for murine studies (Brouwer et al, 2021b; Gristick et al, 2022), whereas 5 μg is lower than previous studies of mRNA-expressed viral antigens (Laczkó et al, 2020; Melzi et al, 2022; Willis et al, 2022). The most efficient neutralizing response was observed when mice were boosted at 3-week intervals ( Fig.…”

Section: Resultsmentioning

confidence: 91%

“…Thus, efficient and physiologically relevant in vivo systems for measuring anti-apex responses will be critical for developing better Env antigens. However, current wild-type rodent, rabbit, or macaque models are not optimal in large part because the diversity (D) gene segments, key contributors to the HCDR3, are highly species-specific (Lefranc, 2014;Lefranc et al, 2015).Transgenic mice can be engineered to express mature or progenitor apex bnAbs (Crooks et al, 2021;Melzi et al, 2022), but these mice are slow to generate or modify, and their antigen reactive repertoires are essentially monoclonal, biasing antigen comparisons. Strategies to engineer mature murine B cells to express human bnAbs and adoptively transfer these cells into wildtype mice have been developed for novel cell-based therapies (Voss et al, 2017;Hartweger et al, 2019;Moffett et al, 2019;Voss et al, 2019;Huang et al, 2020;Nahmad et al, 2020;Nahmad et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

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Heavy-chain CDR3-engineered B cells facilitatein vivoevaluation of HIV-1 vaccine candidates

Wang

Skamangas

et al. 2022

Preprint

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V2-glycan/apex broadly neutralizing antibodies (bnAbs) recognize a closed quaternary epitope of the HIV-1 envelope glycoprotein (Env). This closed structure is necessary to elicit apex antibodies and useful to guide maturation of other bnAb classes. To compare antigens designed to maintain this conformation, apex-specific responses were monitored in mice engrafted with a diverse repertoire of B cells expressing the HCDR3 of the apex bnAb VRC26.25. Engineered B cells affinity matured, guiding improvement of VRC26.25 itself. We found that soluble Env (SOSIP) variants differed significantly in their ability to raise anti-apex responses. A transmembrane SOSIP (SOSIP-TM) delivered as an mRNA-lipid nanoparticle elicited more potent neutralizing responses than multimerized SOSIP proteins. Importantly, SOSIP-TM elicited neutralizing sera from B cells engineered with the predicted VRC26.25-HCDR3 progenitor, which also affinity matured. Our data show that HCDR3-edited B cells facilitate efficient in vivo comparisons of Env antigens and highlight the potential of an HCDR3-focused vaccine approach.

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Section: Resultsmentioning

confidence: 91%

Section: Introductionmentioning

confidence: 99%

Heavy-chain CDR3-engineered B cells facilitatein vivoevaluation of HIV-1 vaccine candidates

Wang

Skamangas

et al. 2022

Preprint

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“…Having established GT5 trimer protein as a robust activator of BG18 gH precursors, we sought to determine whether it could be effectively delivered as a membrane-anchored mRNA. Using a membrane-bound gp151 trimer format reported previously ( 35 , 52 ), we performed immunization experiments as above with 10 μg GT5 membrane-bound trimer mRNA (GT5-mRNA) and analyzed responses at 14, 28, and 56 dpi (Fig. 5A).…”

Section: Resultsmentioning

confidence: 99%

“…Conceptually, GT immunogens should progressively resemble native Env (13), but which immunogens best accommodate binding to the evolving precursor set and drive antibodies along a path to the mature-like state while avoiding undesirable responses is an empirical question. Humanized mouse models have proven critical to the preclinical development of GT immunogens (11,16,30,(32)(33)(34)(35)(36)(37)(38), and we have developed a CRISPR-Cas9 method to rapidly generate mice with B cells expressing human immunoglobulin (Ig) V(D)J sequences (39). These models provided support for the use of N332-GT2 as a potential priming immunogen (19); here, we use them to preclinically validate a next-generation prime, N332-GT5 (referred to below as GT5).…”

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confidence: 99%

mRNA-LNP HIV-1 trimer boosters elicit precursors to broad neutralizing antibodies

Xie,

Lin,

Steichen

et al. 2024

Science

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Germline-targeting (GT) HIV vaccine strategies are predicated on deriving broadly neutralizing antibodies (bnAbs) through multiple boost immunogens. However, as the recruitment of memory B cells (MBCs) to germinal centers (GCs) is inefficient and may be derailed by serum antibody–induced epitope masking, driving further B cell receptor (BCR) modification in GC-experienced B cells after boosting poses a challenge. Using humanized immunoglobulin knockin mice, we found that GT protein trimer immunogen N332-GT5 could prime inferred-germline precursors to the V3-glycan–targeted bnAb BG18 and that B cells primed by N332-GT5 were effectively boosted by either of two novel protein immunogens designed to have minimum cross-reactivity with the off-target V1-binding responses. The delivery of the prime and boost immunogens as messenger RNA lipid nanoparticles (mRNA-LNPs) generated long-lasting GCs, somatic hypermutation, and affinity maturation and may be an effective tool in HIV vaccine development.

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“…Core-C4b: NCT05471076; BG505 SOSIP.GT1.1 gp140: NCT04224701; and eOD-GT8 60mer: NCT03547245 (11)] are designed around VRC01-class bnAbs targeting the Env CD4-binding site (CD4bs) (12)(13)(14)(15). Mice expressing human antibodies have been key to preclinical testing of GT vaccine candidates (6,8,10,(16)(17)(18)(19)(20)(21)(22)(23)(24), including for CD4bs (6, 17-19, 21, 24). We recently developed a CRISPR-Cas9 method to generate mice bearing human B cell receptors (BCRs) (25,26).…”

Section: Introductionmentioning

confidence: 99%

mRNA-LNP prime boost evolves precursors toward VRC01-like broadly neutralizing antibodies in preclinical humanized mouse models

Wang,

Cottrell,

et al. 2024

Sci. Immunol.

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Germline-targeting (GT) protein immunogens to induce VRC01-class broadly neutralizing antibodies (bnAbs) to the CD4-binding site of the HIV envelope (Env) have shown promise in clinical trials. Here, we preclinically validated a lipid nanoparticle–encapsulated nucleoside mRNA (mRNA-LNP) encoding eOD-GT8 60mer as a soluble self-assembling nanoparticle in mouse models. In a model with three humanized B cell lineages bearing distinct VRC01-precursor B cell receptors (BCRs) with similar affinities for eOD-GT8, all lineages could be simultaneously primed and undergo diversification and affinity maturation without exclusionary competition. Boosts drove precursor B cell participation in germinal centers; the accumulation of somatic hypermutations, including in key VRC01-class positions; and affinity maturation to boost and native-like antigens in two of the three precursor lineages. We have preclinically validated a prime-boost regimen of soluble self-assembling nanoparticles encoded by mRNA-LNP, demonstrating that multiple lineages can be primed, boosted, and diversified along the bnAb pathway.

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Membrane-bound mRNA immunogens lower the threshold to activate HIV Env V2 apex-directed broadly neutralizing B cell precursors in humanized mice

Cited by 28 publications

References 90 publications

Heavy-chain CDR3-engineered B cells facilitatein vivoevaluation of HIV-1 vaccine candidates

Heavy-chain CDR3-engineered B cells facilitatein vivoevaluation of HIV-1 vaccine candidates

mRNA-LNP HIV-1 trimer boosters elicit precursors to broad neutralizing antibodies

mRNA-LNP prime boost evolves precursors toward VRC01-like broadly neutralizing antibodies in preclinical humanized mouse models

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