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Author Notes:

Address correspondence and reprint requests to: David M. Koelle, MD, University of Washington, 750 Republican Street, Room E651, Seattle, Washington 98109 USA, Phone 206 616 1940, Fax 206 616 4984, viralimm@u.washington.edu.

K. Nayak and L. Jing contributed equally as first authors.

M.-K.K. and D.M.K. contributed equally as senior authors.

We wish to thank Vu Huynh and Andy Teng (Antigen Discovery, Inc.) for help with protein quality control by microarray printing, and Rick Lawlor for cytokine ELISAs at Fred Hutchinson Cancer Research Center.

The National Institute of Immunology, Delhi, India, kindly allowed use of their cell irradiator.

Subjects:

Research Funding:

Cloning of the ORFS in the Mycobacterium tuberculosis genome was supported by the Foundation of Innovative New Diagnostics (FIND) and by the Bill and Melinda Gates Foundation.

Mycobacterium tuberculosis peptides were kindly provided by BEI Resources, funded by the NIAID at the U.S. National Institutes of Health.

This work was partially supported by the U.S. Public Health Services, National Institutes of Health RO1094019 (D.M.K.), R44 AI58365 (D.H.D.), and HHSN272200900043C (W.W.K.).

Keywords:

  • Life Sciences & Biomedicine
  • Immunology
  • Microbiology
  • Respiratory System
  • Tuberculosis
  • CD4
  • T-cell
  • Antigen
  • CD137
  • Malate synthase
  • Tetramer
  • HUMORAL IMMUNE-RESPONSES
  • ANTIBODY-RESPONSES
  • MALATE SYNTHASE
  • FRANCISELLA-TULARENSIS
  • RAPID IDENTIFICATION
  • SYNTHETIC PEPTIDES
  • INTERFERON-GAMMA

Identification of novel Mycobacterium tuberculosis CD4 T-cell antigens via high throughput proteome screening

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Journal Title:

Tuberculosis

Volume:

Volume 95, Number 3

Publisher:

, Pages 275-287

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Elicitation of CD4 IFN-gamma T cell responses to Mycobacterium tuberculosis (MTB) is a rational vaccine strategy to prevent clinical tuberculosis. Diagnosis of MTB infection is based on T-cell immune memory to MTB antigens. The MTB proteome contains over four thousand open reading frames (ORFs). We conducted a pilot antigen identification study using 164 MTB proteins and MTB-specific T-cells expanded in vitro from 12 persons with latent MTB infection. Enrichment of MTB-reactive T-cells from PBMC used cell sorting or an alternate system compatible with limited resources. MTB proteins were used as single antigens or combinatorial matrices in proliferation and cytokine secretion readouts. Overall, our study found that 44 MTB proteins were antigenic, including 27 not previously characterized as CD4 T-cell antigens. Antigen truncation, peptide, NTM homology, and HLA class II tetramer studies confirmed malate synthase G (encoded by gene Rv1837) as a CD4 T-cell antigen. This simple, scalable system has potential utility for the identification of candidate MTB vaccine and biomarker antigens.

Copyright information:

© 2015 Elsevier Ltd.

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