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

Neelam Dhiman, Inna G. Ovsyannikova, Rawleigh C. Howe, Jenna E. Ryan, Robert M. Jacobson, Gregory A. Poland

Research output: Contribution to journalArticle

23 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)217-225
Number of pages9
JournalJournal of Immunological Methods
Volume287
Issue number1-2
DOIs
StatePublished - Apr 2004

Fingerprint

Interleukin-4 Receptors
Measles virus
Measles
Interleukin-4
Enzyme-Linked Immunosorbent Assay
Peptides
Blood Cells
Measles-Mumps-Rubella Vaccine
Blocking Antibodies
Phosphoproteins
Histocompatibility Antigens Class II
Healthy Volunteers
Cell Culture Techniques
Cytokines
Antigens

Keywords

  • ELISA
  • ELISpot
  • Enzyme-linked immunosorbent assay
  • IFN-γ
  • IL-4
  • IL-4R
  • Interferon-gamma
  • Interleukin-4
  • Interleukin-4 receptors
  • LPA
  • Lymphoproliferative assay
  • Mass spectrometry
  • Measles
  • Measles-mumps-rubella-II
  • MMR-II
  • MS
  • MV
  • PBMC

ASJC Scopus subject areas

  • Biotechnology
  • Immunology

Cite this

Dhiman, N., Ovsyannikova, I. G., Howe, R. C., Ryan, J. E., Jacobson, R. M., & Poland, G. A. (2004). Interleukin-4 induced by measles virus and measles-derived peptides as measured by IL-4 receptor-blocking ELISA. Journal of Immunological Methods, 287(1-2), 217-225. https://doi.org/10.1016/j.jim.2004.02.002

Interleukin-4 induced by measles virus and measles-derived peptides as measured by IL-4 receptor-blocking ELISA. / Dhiman, Neelam; Ovsyannikova, Inna G.; Howe, Rawleigh C.; Ryan, Jenna E.; Jacobson, Robert M.; Poland, Gregory A.

In: Journal of Immunological Methods, Vol. 287, No. 1-2, 04.2004, p. 217-225.

Research output: Contribution to journalArticle

Dhiman, Neelam ; Ovsyannikova, Inna G. ; Howe, Rawleigh C. ; Ryan, Jenna E. ; Jacobson, Robert M. ; Poland, Gregory A. / Interleukin-4 induced by measles virus and measles-derived peptides as measured by IL-4 receptor-blocking ELISA. In: Journal of Immunological Methods. 2004 ; Vol. 287, No. 1-2. pp. 217-225.
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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.",
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T1 - Interleukin-4 induced by measles virus and measles-derived peptides as measured by IL-4 receptor-blocking ELISA

AU - Dhiman, Neelam

AU - Ovsyannikova, Inna G.

AU - Howe, Rawleigh C.

AU - Ryan, Jenna E.

AU - Jacobson, Robert M.

AU - Poland, Gregory A.

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N2 - 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.

AB - 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.

KW - ELISA

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KW - Interferon-gamma

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KW - Interleukin-4 receptors

KW - LPA

KW - Lymphoproliferative assay

KW - Mass spectrometry

KW - Measles

KW - Measles-mumps-rubella-II

KW - MMR-II

KW - MS

KW - MV

KW - PBMC

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