A broadly applicable approach to T cell epitope identification: Application to improving tumor associated epitopes and identifying epitopes in complex pathogens

Michael D. Valentino, C. Siddiq Abdul-Alim, Zachary J. Maben, Denise Skrombolas, Lucinda L. Hensley, Thomas H. Kawula, Michelle Dziejman, Edith M. Lord, Jeffrey A Frelinger, John G. Frelinger

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Epitopes are a hallmark of the antigen specific immune response. The identification and characterization of epitopes is essential for modern immunologic studies, from investigating cellular responses against tumors to understanding host/pathogen interactions especially in the case of bacteria with intracellular residence. Here, we have utilized a novel approach to identify T cell epitopes exploiting the exquisite ability of particulate antigens, in the form of beads, to deliver exogenous antigen to both MHC class I and class II pathways for presentation to T cell hybridomas. In the current study, we coupled this functional assay with two distinct protein expression libraries to develop a methodology for the characterization of T cell epitopes. One set of expression libraries containing single amino acid substitutions in a defined epitope sequence was interrogated to identify epitopes with enhanced T cell stimulation for a MHC class I epitope. The second expression library is comprised of the majority of open reading frames from the intracellular pathogen and potential biowarfare agent, Francisella tularensis. By automating aspects of this technology, we have been able to functionally screen and identify novel T cell epitopes within F. tularensis. We have also expanded upon these studies to generate a novel expression vector that enables immunization of recombinant protein into mice, which has been utilized to facilitate T cell epitope discovery for proteins that are critically linked to Francisella pathogenicity. This methodology should be applicable to a variety of systems and other pathogens.

Original languageEnglish (US)
Pages (from-to)111-126
Number of pages16
JournalJournal of Immunological Methods
Volume373
Issue number1-2
DOIs
StatePublished - Oct 28 2011

Fingerprint

T-Lymphocyte Epitopes
Epitopes
Francisella tularensis
Neoplasms
Francisella
Biological Warfare Agents
T-Lymphocytes
Host-Pathogen Interactions
Antigens
Histocompatibility Antigens Class II
Hybridomas
Amino Acid Substitution
Recombinant Proteins
Open Reading Frames
Virulence
Immunization
Proteins
Technology
Bacteria

Keywords

  • Altered peptide ligands
  • Francisella tularensis
  • High-throughput screening
  • Prostate specific antigen
  • Protein expression library
  • T cell epitope discovery

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

A broadly applicable approach to T cell epitope identification : Application to improving tumor associated epitopes and identifying epitopes in complex pathogens. / Valentino, Michael D.; Abdul-Alim, C. Siddiq; Maben, Zachary J.; Skrombolas, Denise; Hensley, Lucinda L.; Kawula, Thomas H.; Dziejman, Michelle; Lord, Edith M.; Frelinger, Jeffrey A; Frelinger, John G.

In: Journal of Immunological Methods, Vol. 373, No. 1-2, 28.10.2011, p. 111-126.

Research output: Contribution to journalArticle

Valentino, Michael D. ; Abdul-Alim, C. Siddiq ; Maben, Zachary J. ; Skrombolas, Denise ; Hensley, Lucinda L. ; Kawula, Thomas H. ; Dziejman, Michelle ; Lord, Edith M. ; Frelinger, Jeffrey A ; Frelinger, John G. / A broadly applicable approach to T cell epitope identification : Application to improving tumor associated epitopes and identifying epitopes in complex pathogens. In: Journal of Immunological Methods. 2011 ; Vol. 373, No. 1-2. pp. 111-126.
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