Peanut T-cell epitope discovery

Ara h 1

Manish Ramesh, Araya Yuenyongviwat, George N. Konstantinou, Jay Lieberman, Mariona Pascal, Madhan Masilamani, Hugh A. Sampson

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background Identification of potential T-cell epitopes in the peanut major allergens is essential for development of peptide-based immunotherapy. Traditional methods of T-cell epitope discovery use overlapping short peptides spanning the full length of the protein in T-cell proliferation assays. Because large proteins, such as Ara h 1, require a large number of peptides, this limits screening to a small number of allergic subject-derived T-cell lines. Objective We sought to identify candidate peptides of Ara h 1 that display promiscuous binding to MHC class II and induce T H 2 cytokine production by T cells. Methods In silico MHC class II binding prediction was performed with NetMHCIIpan 2.0 (peptide length, 15; 1-mer offset) and the most abundant class II alleles in the North American population and with an in vitro MHC class II peptide reporter assay performed in parallel, which used synthetic 15-mer peptides offset by 5 mer spanning the protein. High-resolution MHC class II typing and a T-cell proliferation assay using preselected peptides were performed with PBMCs from 98 subjects with peanut allergy and 14 healthy control subjects. IL-4, IL-13, IL-5, IFN-γ, and TNF-α levels were measured in culture supernatants. Results Thirty-six Ara h 1 peptides were identified by using in silico predictions and MHC class II binding assays. In combination with T-cell proliferation and cytokines secreted in T-cell assays, we have identified 4 vaccine candidate Ara h 1 peptides. Conclusions Preselection of peptides by using in silico and in vitro approaches in combination with conventional methods appears to be an effective strategy for identifying peanut T-cell peptide vaccine candidates.

Original languageEnglish (US)
Pages (from-to)1764-1771.e4
JournalJournal of Allergy and Clinical Immunology
Volume137
Issue number6
DOIs
StatePublished - Jun 1 2016

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T-Lymphocyte Epitopes
Peptides
T-Lymphocytes
Computer Simulation
Cell Proliferation
Arachis
Peanut Hypersensitivity
Cytokines
Subunit Vaccines
Proteins
Interleukin-13
Interleukin-5
Interleukin-4
Immunotherapy
Allergens
Healthy Volunteers
Vaccines
Alleles

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Ramesh, M., Yuenyongviwat, A., Konstantinou, G. N., Lieberman, J., Pascal, M., Masilamani, M., & Sampson, H. A. (2016). Peanut T-cell epitope discovery: Ara h 1. Journal of Allergy and Clinical Immunology, 137(6), 1764-1771.e4. https://doi.org/10.1016/j.jaci.2015.12.1327

Peanut T-cell epitope discovery : Ara h 1. / Ramesh, Manish; Yuenyongviwat, Araya; Konstantinou, George N.; Lieberman, Jay; Pascal, Mariona; Masilamani, Madhan; Sampson, Hugh A.

In: Journal of Allergy and Clinical Immunology, Vol. 137, No. 6, 01.06.2016, p. 1764-1771.e4.

Research output: Contribution to journalArticle

Ramesh, M, Yuenyongviwat, A, Konstantinou, GN, Lieberman, J, Pascal, M, Masilamani, M & Sampson, HA 2016, 'Peanut T-cell epitope discovery: Ara h 1', Journal of Allergy and Clinical Immunology, vol. 137, no. 6, pp. 1764-1771.e4. https://doi.org/10.1016/j.jaci.2015.12.1327
Ramesh M, Yuenyongviwat A, Konstantinou GN, Lieberman J, Pascal M, Masilamani M et al. Peanut T-cell epitope discovery: Ara h 1. Journal of Allergy and Clinical Immunology. 2016 Jun 1;137(6):1764-1771.e4. https://doi.org/10.1016/j.jaci.2015.12.1327
Ramesh, Manish ; Yuenyongviwat, Araya ; Konstantinou, George N. ; Lieberman, Jay ; Pascal, Mariona ; Masilamani, Madhan ; Sampson, Hugh A. / Peanut T-cell epitope discovery : Ara h 1. In: Journal of Allergy and Clinical Immunology. 2016 ; Vol. 137, No. 6. pp. 1764-1771.e4.
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abstract = "Background Identification of potential T-cell epitopes in the peanut major allergens is essential for development of peptide-based immunotherapy. Traditional methods of T-cell epitope discovery use overlapping short peptides spanning the full length of the protein in T-cell proliferation assays. Because large proteins, such as Ara h 1, require a large number of peptides, this limits screening to a small number of allergic subject-derived T-cell lines. Objective We sought to identify candidate peptides of Ara h 1 that display promiscuous binding to MHC class II and induce T H 2 cytokine production by T cells. Methods In silico MHC class II binding prediction was performed with NetMHCIIpan 2.0 (peptide length, 15; 1-mer offset) and the most abundant class II alleles in the North American population and with an in vitro MHC class II peptide reporter assay performed in parallel, which used synthetic 15-mer peptides offset by 5 mer spanning the protein. High-resolution MHC class II typing and a T-cell proliferation assay using preselected peptides were performed with PBMCs from 98 subjects with peanut allergy and 14 healthy control subjects. IL-4, IL-13, IL-5, IFN-γ, and TNF-α levels were measured in culture supernatants. Results Thirty-six Ara h 1 peptides were identified by using in silico predictions and MHC class II binding assays. In combination with T-cell proliferation and cytokines secreted in T-cell assays, we have identified 4 vaccine candidate Ara h 1 peptides. Conclusions Preselection of peptides by using in silico and in vitro approaches in combination with conventional methods appears to be an effective strategy for identifying peanut T-cell peptide vaccine candidates.",
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N2 - Background Identification of potential T-cell epitopes in the peanut major allergens is essential for development of peptide-based immunotherapy. Traditional methods of T-cell epitope discovery use overlapping short peptides spanning the full length of the protein in T-cell proliferation assays. Because large proteins, such as Ara h 1, require a large number of peptides, this limits screening to a small number of allergic subject-derived T-cell lines. Objective We sought to identify candidate peptides of Ara h 1 that display promiscuous binding to MHC class II and induce T H 2 cytokine production by T cells. Methods In silico MHC class II binding prediction was performed with NetMHCIIpan 2.0 (peptide length, 15; 1-mer offset) and the most abundant class II alleles in the North American population and with an in vitro MHC class II peptide reporter assay performed in parallel, which used synthetic 15-mer peptides offset by 5 mer spanning the protein. High-resolution MHC class II typing and a T-cell proliferation assay using preselected peptides were performed with PBMCs from 98 subjects with peanut allergy and 14 healthy control subjects. IL-4, IL-13, IL-5, IFN-γ, and TNF-α levels were measured in culture supernatants. Results Thirty-six Ara h 1 peptides were identified by using in silico predictions and MHC class II binding assays. In combination with T-cell proliferation and cytokines secreted in T-cell assays, we have identified 4 vaccine candidate Ara h 1 peptides. Conclusions Preselection of peptides by using in silico and in vitro approaches in combination with conventional methods appears to be an effective strategy for identifying peanut T-cell peptide vaccine candidates.

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