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Mapping the epitope of neutralizing monoclonal antibodies against human adenovirus type 3
Xingui Tian1, Minglong Liu1, Xiaobo Su2
1State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, The Affiliated First Hospital of Guangzhou Medical University, Guangzhou 510230, China.
Abstract:
Human adenovirus type 3 (HAdV-3) has produced a global epidemic in recent years causing serious diseases such as pneumonia in both pediatric and adult patients. Development of an effective neutralizing monoclonal antibody (MAb) and identification of its neutralizing epitope is important for the control of HAdV-3 infection. In this study, three neutralizing MAbs were generated, of which MAb 3D7 had a high neutralization titer of 4096 (approximately 0.5 μg/ml) against HAdV-3 infection. In indirect enzyme-linked immunosorbent assays, all three MAbs specifically recognized HAdV-3 virus particles and hexon protein, but did not react with the virus particles or the hexon protein of HAdV-7. Analyses using a series of peptides and chimeric adenovirus particles of epitope mutants revealed that all three MAbs bound to the same exposed region (amino acid positions 244-254 of hexon) in hypervariable region 4 (HVR4), which is highly conserved among global HAdV-3 strains. The amino acids T246 and G250 may be the critical amino acids recognized by these MAbs. MAb 3D7 reduced the recombinant enhanced green fluorescent protein-expressing HAdV-3 (rAd3EGFP) load recovered in the lungs of mice at 3 days post-infection. The generation of MAb 3D7 and the identification of its neutralizing epitope may be useful for therapeutic treatment development, subunit vaccine construction, and virion structural analysis for HAdV-3.
Insights
Researchers developed a neutralizing antibody, MAb 3D7, effective against Human adenovirus type 3 (HAdV-3) infections. The antibody targets a specific epitope on the HAdV-3 hexon protein, showing potential for therapeutic development and HAdV-3 control.
Area of Science:
- Virology
- Immunology
- Structural Biology
Background:
- Human adenovirus type 3 (HAdV-3) causes global epidemics, leading to severe pneumonia in children and adults.
- Effective control strategies, including neutralizing monoclonal antibodies (MAbs), are crucial for managing HAdV-3 infections.
Purpose of the Study:
- To generate neutralizing MAbs against HAdV-3.
- To identify the neutralizing epitope targeted by these MAbs.
- To evaluate the therapeutic potential of a lead MAb in a mouse model.
Main Methods:
- Generation and characterization of neutralizing MAbs against HAdV-3.
- Indirect enzyme-linked immunosorbent assays (ELISAs) for specificity testing.
- Epitope mapping using peptides and chimeric adenovirus particles.
- In vivo efficacy study in a mouse model of HAdV-3 infection.
Main Results:
- Three neutralizing MAbs were generated, with MAb 3D7 showing a high neutralization titer (4096).
- MAbs specifically recognized HAdV-3 virus and hexon protein, not HAdV-7.
- Epitope mapping identified a conserved region (amino acids 244-254 in HVR4) on the hexon protein.
- MAb 3D7 significantly reduced viral load in the lungs of infected mice.
Conclusions:
- MAb 3D7 is a potent neutralizing antibody against HAdV-3.
- The identified epitope on the HAdV-3 hexon protein is a key target for neutralization.
- MAb 3D7 demonstrates therapeutic potential for HAdV-3 infections and can aid in vaccine development.
