Identification of broad-spectrum neutralizing antibodies against influenza A virus and evaluation of their prophylactic efficacy in mice

Son, Sumin; Ahn, Soo Bin; Kim, Geonyeong; Jang, Yejin; Ko, Chunkyu; Kim, Meehyein; Kim, Sang Jick

doi:10.1016/j.antiviral.2023.105591

Cited by 3 publications

(4 citation statements)

References 56 publications

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“…To produce S2P6 antibody [ 34 ] in the form of human IgG1, the sequences for the variable regions of both the heavy and light chains (Integrated DNA Technologies, Coralville, IA, USA) were synthetized. These sequences were then inserted into the pCEP-WPRE-hCH (hIgG1) vector for the heavy chain and the pCEP-WPRE-hCK vector for the light chain using the NEBuilder HiFi DNA Assembly system (New England Biolabs, Ipswich, MA, USA), following the protocols previously outlined [ 42 ]. After constructing the vectors for both chains, they were co-transfected into ExpiCHO cells (Thermo Fisher Scientific, Waltham, MA, USA).…”

Section: Methodsmentioning

confidence: 99%

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S2 Peptide-Conjugated SARS-CoV-2 Virus-like Particles Provide Broad Protection against SARS-CoV-2 Variants of Concern

Heo,

Lim,

Moon

et al. 2024

Vaccines

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Approved COVID-19 vaccines primarily induce neutralizing antibodies targeting the receptor-binding domain (RBD) of the SARS-CoV-2 spike (S) protein. However, the emergence of variants of concern with RBD mutations poses challenges to vaccine efficacy. This study aimed to design a next-generation vaccine that provides broader protection against diverse coronaviruses, focusing on glycan-free S2 peptides as vaccine candidates to overcome the low immunogenicity of the S2 domain due to the N-linked glycans on the S antigen stalk, which can mask S2 antibody responses. Glycan-free S2 peptides were synthesized and attached to SARS-CoV-2 virus-like particles (VLPs) lacking the S antigen. Humoral and cellular immune responses were analyzed after the second booster immunization in BALB/c mice. Enzyme-linked immunosorbent assay revealed the reactivity of sera against SARS-CoV-2 variants, and pseudovirus neutralization assay confirmed neutralizing activities. Among the S2 peptide-conjugated VLPs, the S2.3 (N1135-K1157) and S2.5 (A1174-L1193) peptide–VLP conjugates effectively induced S2-specific serum immunoglobulins. These antisera showed high reactivity against SARS-CoV-2 variant S proteins and effectively inhibited pseudoviral infections. S2 peptide-conjugated VLPs activated SARS-CoV-2 VLP-specific T-cells. The SARS-CoV-2 vaccine incorporating conserved S2 peptides and CoV-2 VLPs shows promise as a universal vaccine capable of generating neutralizing antibodies and T-cell responses against SARS-CoV-2 variants.

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Section: Methodsmentioning

confidence: 99%

“…After constructing the vectors for both chains, they were co-transfected into ExpiCHO cells (Thermo Fisher Scientific, Waltham, MA, USA). The recombinant IgG molecules were subsequently produced and purified according to the referenced protocol [ 42 ].…”

Section: Methodsmentioning

confidence: 99%

S2 Peptide-Conjugated SARS-CoV-2 Virus-like Particles Provide Broad Protection against SARS-CoV-2 Variants of Concern

Heo,

Lim,

Moon

et al. 2024

Vaccines

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“…For human IgG1 expression of S2P6 ( 17 ), variable region sequences of heavy and light chains were synthesized (Integrated DNA Technologies, Coralville, IA, USA) and cloned into pCEP-WPRE-hCH (hIgG1) and pCEP-WPRE-hCK for heavy and light chains, respectively, via the NEBuilder HiFi DNA Assembly system (New England Biolabs, Ipswich, MA, USA). Heavy and light chain constructs were cotransfected into ExpiCHO cells (Thermo Fisher Scientific, Waltham, MA, USA), and recombinant IgG molecules were produced and purified as previously described ( 18 ). The following antibodies were used: anti-S2 mAbs NBP2 (Novus Biologicals, Centennial, CO, USA, Cat#NBP2-90999) and MA5 (Thermo Fisher Scientific, Cat#MA5-35946), anti-RBD mAb MM43 (Sino Biological, Cat# 40591-MM43) and anti-SARS-CoV-2 NP mAb 1G6 ( 19 ).…”

Section: Methodsmentioning

confidence: 99%

Novel S2 subunit-specific antibody with broad neutralizing activity against SARS-CoV-2 variants of concern

Heo,

Lim,

Yang

et al. 2023

Front. Immunol.

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IntroductionSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), had a major impact on both the global health and economy. Numerous virus-neutralizing antibodies were developed against the S1 subunit of SARS-CoV-2 spike (S) protein to block viral binding to host cells and were authorized for control of the COVID-19 pandemic. However, frequent mutations in the S1 subunit of SARS-CoV-2 enabled the emergence of immune evasive variants. To address these challenges, broadly neutralizing antibodies targeting the relatively conserved S2 subunit and its epitopes have been investigated as antibody therapeutics and universal vaccines.MethodsWe initiated this study by immunizing BALB/c mice with β-propiolactone-inactivated SARS-CoV-2 (IAV) to generate B-cell hybridomas. These hybridomas were subsequently screened using HEK293T cells expressing the S2-ECD domain. Hybridomas that produced anti-S2 antibodies were selected, and we conducted a comprehensive evaluation of the potential of these anti-S2 antibodies as antiviral agents and versatile tools for research and diagnostics.ResultsIn this study, we present a novel S2-specific antibody, 4A5, isolated from BALB/c mice immunized with inactivated SARS-CoV-2. 4A5 exhibited specific affinity to SARS-CoV-2 S2 subunits compared with those of other β-CoVs. 4A5 bound to epitope segment F1109–V1133 between the heptad-repeat1 (HR1) and the stem-helix (SH) region. The 4A5 epitope is highly conserved in SARS-CoV-2 variants, with a significant conformational feature in both pre- and postfusion S proteins. Notably, 4A5 exhibited broad neutralizing activity against variants and triggered Fc-enhanced antibody-dependent cellular phagocytosis.DiscussionThese findings offer a promising avenue for novel antibody therapeutics and insights for next-generation vaccine design. The identification of 4A5, with its unique binding properties and broad neutralizing capacity, offers a potential solution to the challenge posed by SARS-CoV-2 variants and highlights the importance of targeting the conserved S2 subunit in combating the COVID-19.

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“…HA consists of two subunits: HA1 and HA2; these subunits play different roles in the onset of the infectious process. A number of antibodies that specifically interact with H3 HA and the entire phylogenetic group 2 through different mechanisms have been described [ 16 , 17 , 18 , 19 , 20 , 1 , 22 , 23 , 24 , 25 , 26 , 27 ]. The Fc-mediated antibody function is one of the mechanisms involved in combating an influenza infection [ 28 , 29 ].…”

Section: Introductionmentioning

confidence: 99%

Broadly Reactive Nanobody Targeting the H3 Hemagglutinin of the Influenza A Virus

Shcheblyakov,

Voronina,

Favorskaya

et al. 2024

Acta Naturae

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Monoclonal antibodies and recombinant antibody fragments are a very promising therapeutic tool to combat infectious diseases. Due to their unique paratope structure, nanobodies (VHHs) hold several advantages over conventional monoclonal antibodies, especially in relation to viral infections. Influenza A viruses (IAVs) remain a major threat to public health. The hemagglutinin (HA) protein is the main protective and immunodominant antigen of IAVs. In this study, three broadly reactive nanobodies (D9.2, E12.2, and D4.2) to H3N2 influenza strains were isolated and Fc-fusion proteins (VHH-Fcs) were obtained and characterized in vitro. This modification improved the nanobodies’ binding activity and allowed for their interaction with a wider range of strains. The D9.2-Fc antibody showed a 100% protection rate against mortality in vivo in a mouse lethal model. Furthermore, we demonstrated that the observed protection has to do with Fc-FcγR interactions. These results indicate that D9.2-Fc can serve as an effective antiviral agent against the H3N2 influenza infection.

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Identification of broad-spectrum neutralizing antibodies against influenza A virus and evaluation of their prophylactic efficacy in mice

Cited by 3 publications

References 56 publications

S2 Peptide-Conjugated SARS-CoV-2 Virus-like Particles Provide Broad Protection against SARS-CoV-2 Variants of Concern

S2 Peptide-Conjugated SARS-CoV-2 Virus-like Particles Provide Broad Protection against SARS-CoV-2 Variants of Concern

Novel S2 subunit-specific antibody with broad neutralizing activity against SARS-CoV-2 variants of concern

Broadly Reactive Nanobody Targeting the H3 Hemagglutinin of the Influenza A Virus

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