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Updated: Feb 24, 2026

An Optimized Hemagglutination Inhibition HI Assay to Quantify Influenza-specific Antibody Titers
Published on: December 1, 2017
Plasma cell and serum antibody responses to influenza vaccine in preterm and full-term infants
Carl T D'Angio1, Claire P Wyman1, Ravi S Misra1
1Department of Pediatrics, University of Rochester School of Medicine and Dentistry, 601 Elmwood Avenue, Rochester, NY 14642, USA.
Insights
Preterm infants demonstrate robust antibody responses to influenza vaccines, comparable to full-term infants. Higher influenza-specific antibody secreting cell numbers in preterm infants correlate with peak antibody titers, suggesting effective immune responses.
Area of Science:
- Immunology
- Pediatrics
- Vaccinology
Background:
- Preterm (PT) infants face higher risks of severe influenza and diminished vaccine antibody responses.
- This may stem from impaired antibody secreting cell (ASC) generation in PT infants.
Purpose of the Study:
- To compare influenza-specific ASC frequencies, numbers, subsets, and antibody responses to influenza vaccines (IV) between PT and full-term (FT) infants.
- To investigate the relationship between ASCs and antibody kinetics following IV in these populations.
Main Methods:
- 11 PT (≤32 weeks' gestation, ≤1500 g) and 11 FT infants (6-17 months) received two doses of inactivated influenza vaccine.
- Blood samples were collected at multiple time points for antibody (HAI), ASC numbers (ELISPOT), and ASC subsets (flow cytometry) analysis.
Main Results:
- PT infants achieved HAI titers equal to or greater than FT infants at 56 days and 9 months post-immunization.
- Influenza-specific ASC ELISPOT responses were significantly higher in PT infants at 35 days (median 100 vs. 30 per 10^6 PBMC, p=0.04).
- ASC numbers at 35 days positively correlated with serum HAI titers at 56 days (ρ=0.50-0.80).
Conclusions:
- Influenza-specific ASC numbers correlate with peak antibody titers in both PT and FT infants.
- No significant differences in ASC subsets were observed between PT and FT infants.
- ASC subsets did not correlate with the durability of serum antibody response.
Background:
Preterm (PT) infants are at greater risk for severe influenza infection and experience decrements in long-term antibody responses to vaccines. This may related to defects in antibody secreting cell (ASC) generation.
Objective:
To investigate the relationships among the frequencies of influenza-specific antibody secreting cells, ASC numbers and subsets, and antibody responses to influenza vaccines (IV) among PT and full-term (FT) infants.
Design/Methods:
We enrolled 11 former PT (≤32weeks' gestation, ≤1500 g' birth weight) and 11FT infants, 6-17months of age, receiving their first influenza immunizations. Infants received two doses of inactivated trivalent (T)IV or quadrivalent (Q)IV during the 2012-2013 and 2013-2014 influenza seasons, respectively, at 0 and 28days, and blood was drawn at 0, 10, 35, and 56days and 9months. Vaccine-specific antibody was measured by hemagglutination inhibition (HAI) at 0 and 56days and 9months, vaccine-specific ASC numbers by enzyme linked immunospot (ELISPOT) at 10 and 35days, and ASC subsets by flow cytometry at 0, 10 and 35days.
Results:
PT infants had post-vaccine HAI titers to all 4 vaccine strains at least equal to FT infants at 56days and 9months after beginning immunization. Influenza-specific ASC ELISPOT responses at 35days were higher among PT than FT infants (median 100 v. 30 per 106 PBMC, p=0.04). ASC numbers at 35days were positively correlated with serum HAI titers at 56days (ρ=0.50-0.80). There were no statistical differences between PT and FT infants in the frequency of five ASC subsets and no specific ASC subset correlated with durability of serum antibody titers.
Conclusions:
Influenza-specific ASC numbers in both FT and PT infants correlated with peak antibody titers, but ASC subsets did not correlate with durability of antibody response.
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