Reliability of pseudotyped influenza viral particles in neutralizing antibody detection
Jinghui Yang1, Weidong Li1, Yunfeng Long1
1Institute of Medical Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Kunming, Yunnan, China.
Background:
Current influenza control strategies require an active surveillance system. Pseudotyped viral particles (pp) together with the evaluation of pre-existing immunity in a population might satisfy this requirement. However, the reliability of using pp in neutralizing antibody (nAb) detection are undefined.
Methodology/Principal Findings:
Pseudotyped particles of A(H1N1)pmd09 (A/California/7/2009) and HPAI H5N1 (A/Anhui/1/2005), as well as their reassortants, were generated. The reliability of using these pp in nAb detection were compared concurrently with the corresponding viruses by a hemagglutination inhibition test, as well as ELISA-, cytopathic effect-, and fluorescence-based microneutralization assays. In the qualitative detection on nAbs, the pp and their corresponding viruses were in complete agreement, with an R2 value equal to or near 1 in two different populations. In the quantitative detection on nAbs, although the geometric mean titers (95% confidence interval) differed between the pp and viruses, no significant difference was observed. Furthermore, humoral immunity against the reassortants was evaluated; our results indicated strong consistency between the nAbs against reassortant pp and those against naïve pp harboring the same hemagglutinin.
Conclusion/Significance:
The pp displayed high reliability in influenza virus nAb detection. The use of reassortant pp is a safe and convenient strategy for characterizing emerging influenza viruses and surveying the disease burden.
Insights
Pseudotyped viral particles (pp) offer a reliable method for detecting neutralizing antibodies (nAbs) against influenza viruses. This safe and convenient strategy aids in characterizing emerging strains and monitoring disease burden.
Area of Science:
- Virology
- Immunology
- Public Health
Background:
- Active surveillance is crucial for influenza control.
- Pseudotyped viral particles (pp) offer a potential tool for evaluating population immunity.
- The reliability of pp for neutralizing antibody (nAb) detection requires validation.
Purpose of the Study:
- To assess the reliability of pseudotyped viral particles (pp) for detecting neutralizing antibodies (nAbs) against influenza.
- To compare nAb detection using pp with conventional methods and live viruses.
- To evaluate the utility of reassortant pp for characterizing emerging influenza strains.
Main Methods:
- Generated pseudotyped particles (pp) for A(H1N1)pmd09 and HPAI H5N1 strains, including reassortants.
- Compared pp-based nAb detection with hemagglutination inhibition, ELISA, cytopathic effect, and microneutralization assays.
- Assessed qualitative and quantitative nAb detection and humoral immunity against reassortant pp.
Main Results:
- Pseudotyped particles demonstrated complete agreement with live viruses in qualitative nAb detection (R2 ≈ 1).
- Quantitative nAb detection showed no significant differences between pp and live viruses, despite titer variations.
- Strong consistency was observed in nAb detection against reassortant pp and their corresponding naïve counterparts.
Conclusions:
- Pseudotyped viral particles (pp) exhibit high reliability for influenza virus neutralizing antibody (nAb) detection.
- Reassortant pp provide a safe and convenient method for characterizing emerging influenza viruses.
- This approach facilitates effective surveillance of influenza disease burden.
