Abstract
Interleukin-4 (IL-4) is a signature cytokine for T-helper 2 (Th2) type immune responses in humans. However, data on antigen-specific secretion of IL-4 is limited due to difficulties detecting IL-4. We evaluated an IL-4 receptor-blocking assay for the detection of secreted IL-4 in peripheral blood mononuclear cells (PBMC) stimulated in vitro with measles virus (MV) and MV-derived nucleoprotein (N) and phosphoprotein (P) peptides. We recruited 20 healthy subjects, previously immunized with two doses of measles-mumps-rubella- II (MMR-II) vaccine. We evaluated the cellular and humoral immune status of these study subjects by an in vitro lymphoproliferation assay and an enzyme-linked immunosorbent assay (ELISA), respectively. We analyzed the MV-induced and N and P peptide-induced IL-4 levels in PBMC culture supernatants. Using the IL-4 receptor blocking assay, 50% of the subjects were positive for secreted IL-4 in response to MV stimulation, and 5% and 23.1% of study subjects were positive for secreted IL-4 in response to MV-derived N and P peptides, respectively. In contrast, we did not find any positive secreted IL-4 response to MV using conventional ELISA without IL-4 receptor-blocking antibody in our optimization study. Further, we found very low frequencies of IL-4 secreting cells using an alternate ELISpot technique, accounting for only a 5% positive response to MV and no response to P peptide. We propose that the IL-4 receptor-blocking assay is an easy-to-adapt technique for screening antigen-specific immune responses in large-scale population-based studies.
Original language | English (US) |
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Pages (from-to) | 217-225 |
Number of pages | 9 |
Journal | Journal of Immunological Methods |
Volume | 287 |
Issue number | 1-2 |
DOIs | |
State | Published - Apr 2004 |
Keywords
- ELISA
- ELISpot
- Enzyme-linked immunosorbent assay
- IFN-γ
- IL-4
- IL-4R
- Interferon-gamma
- Interleukin-4
- Interleukin-4 receptors
- LPA
- Lymphoproliferative assay
- MMR-II
- MS
- MV
- Mass spectrometry
- Measles
- Measles-mumps-rubella-II
- PBMC
ASJC Scopus subject areas
- Immunology and Allergy
- Immunology