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Updated: Jul 12, 2026
An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection
Published on: December 10, 2013
Development and comparison of immunologic assays to detect primary RSV infections in infants
Larry J Anderson1,2, Samadhan J Jadhao1,2, Laila Hussaini1,2
1Division of Infectious Diseases, Department of Pediatrics, Emory University School of Medicine, Atlanta, GA, United States.
Insights
Sensitive assays are crucial for tracking respiratory syncytial virus (RSV) immunity in children. The study found that lysate and F protein IgG enzyme immunoassays (EIAs) are most effective for detecting past RSV infection in young children.
Area of Science:
- Immunology
- Vaccinology
- Pediatrics
Background:
- Respiratory syncytial virus (RSV) vaccines are available for adults and pregnant women, but not yet for infants and young children.
- Determining a young child's RSV immune status is vital for epidemiological studies and vaccine clinical trials.
- Sensitive assays are needed to detect immunologic evidence of past RSV infection in children.
Purpose of the Study:
- To develop, characterize, and evaluate seven different assays for detecting immunologic evidence of past RSV infection in young children.
- To identify the most sensitive and reliable assays for use in pediatric RSV epidemiology and vaccine research.
Main Methods:
- Developed and evaluated four IgG enzyme immunoassays (EIAs) using lysate, F protein, G protein A, or G protein B antigens.
- Assessed two neutralizing antibody assays (subgroup A and B) and one IFN-γ EliSpot assay.
- Tested assay performance including repeatability, limit of detection, and interference from sample conditions.
Main Results:
- In children with documented RSV illness, lysate and F protein IgG EIAs showed 100% sensitivity.
- Neutralizing antibody assays had lower sensitivity (43-57%), while G protein EIAs varied (86-95%).
- The EliSpot assay showed unexpected positivity in infants without recent RSV exposure, possibly due to maternal microchimerism.
Conclusions:
- Lysate and F protein IgG EIAs are highly sensitive and reliable for detecting RSV antibodies in young children.
- These assays are suitable for use in pediatric RSV epidemiologic studies and vaccine trials.
- Further investigation into the EliSpot assay's reactivity in infants may be warranted.
Abstract:
Effective respiratory syncytial virus (RSV) vaccines have been developed and licensed for elderly adults and pregnant women but not yet for infants and young children. The RSV immune state of the young child, i.e., previously RSV infected or not, is important to the conduct and interpretation of epidemiology studies and vaccine clinical trials. To address the need for sensitive assays to detect immunologic evidence of past infection, we developed, characterized, and evaluated 7 assays including 4 IgG antibody enzyme immunoassays (EIAs), two neutralizing antibody assays, and an IFN-γ EliSpot (EliSpot) assay. The four IgG EIAs used a subgroup A plus subgroup B RSV-infected Hep-2 cell lysate antigen (Lysate), an expressed RSV F protein antigen (F), an expressed subgroup A G protein antigen (Ga), or an expressed subgroup B G protein (Gb) antigen. The two neutralizing antibody assays used either a subgroup A or a subgroup B RSV strain. The EliSpot assay used a sucrose cushion purified combination of subgroup A and subgroup B infected cell lysate. All seven assays had acceptable repeatability, signal against control antigen, lower limit of detection, and, for the antibody assays, effect of red cell lysis, lipemia and anticoagulation of sample on results. In 44 sera collected from children >6 months after an RSV positive illness, the lysate, F, Ga and Gb IgG EIAs, and the subgroup A and B neutralizing antibody assays, and the EliSpot assays were positive in 100%, 100%, 86%, 95%, 43%, and 57%, respectively. The Lysate and F EIAs were most sensitive for detecting RSV antibody in young children with a documented RSV infection. Unexpectedly, the EliSpot assay was positive in 9/15 (60%) of PBMC specimens from infants not exposed to an RSV season, possibly from maternal microchimerism. The Lysate and F EIAs provide good options to reliably detect RSV antibodies in young children for epidemiologic studies and vaccine trials.
