Interleukin-4 induced by measles virus and measles-derived peptides as measured by IL-4 receptor-blocking ELISA.
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.
Dhiman
Neelam
N
Mayo Vaccine Research Group, Mayo Clinic and Foundation, Rochester, MN 55905, USA.
Ovsyannikova
Inna G
IG
Howe
Rawleigh C
RC
Ryan
Jenna E
JE
Jacobson
Robert M
RM
Poland
Gregory A
GA
D000293:Adolescent; D000956:Antigens, Viral / Q000276:immunology*; D002478:Cells, Cultured; D002648:Child; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D015847:Interleukin-4 / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology* / Q000378:metabolism; D008297:Male; D008403:Mass Screening; D008459:Measles virus / Q000276:immunology*; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000276:immunology; D010750:Phosphoproteins / Q000276:immunology; D019948:Receptors, Interleukin-4 / Q000276:immunology*; D014764:Viral Proteins / Q000276:immunology
D000956:Antigens, Viral; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D010750:Phosphoproteins; D019948:Receptors, Interleukin-4; D014764:Viral Proteins; C058314:nucleoprotein, Measles virus; D015847:Interleukin-4; D007371:Interferon-gamma