Interleukin-4 induced by measles virus and measles-derived peptides as measured by IL-4 receptor-blocking ELISA

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.