title: Identification of novel Mycobacterium tuberculosis CD4 T-cell antigens via high throughput proteome screening.
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.
authors: ICGEB-Emory Vaccine Center, International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India. Electronic address: kaustuvnayak@gmail.com.
authors: Kaustuv
authors: K
authors: Nayak
authors: Department of Medicine, Division of Infectious Diseases, University of Washington, Box 358061, Seattle, WA 98195, USA. Electronic address: lchjing@u.washington.edu.
authors: Lichen
authors: L
authors: Jing
authors: Department of Medicine, Division of Infectious Diseases, University of Washington, Box 358061, Seattle, WA 98195, USA. Electronic address: ronnie53@u.washington.edu.
authors: Ronnie M
authors: RM
authors: Russell
authors: Department of Medicine, Division of Infectious Diseases, University of California, Room 376D Med-Surg II, Irvine, CA 92697-4068, USA; Antigen Discovery, Inc., 1 Technology Drive Suite E309, Irvine, CA 92618, USA. Electronic address: ddavies@uci.edu.
authors: D Huw
authors: DH
authors: Davies
authors: Antigen Discovery, Inc., 1 Technology Drive Suite E309, Irvine, CA 92618, USA. Electronic address: ghermason@antigendiscovery.com.
authors: Gary
authors: G
authors: Hermanson
authors: Antigen Discovery, Inc., 1 Technology Drive Suite E309, Irvine, CA 92618, USA. Electronic address: dmolina@antigendiscovery.com.
authors: Douglas M
authors: DM
authors: Molina
authors: Antigen Discovery, Inc., 1 Technology Drive Suite E309, Irvine, CA 92618, USA. Electronic address: xliang@immport-inc.com.
authors: Xiaowu
authors: X
authors: Liang
authors: Seattle Biomedical Research Institute, 307 Westlake Ave. North, No. 500, Seattle, WA 98109, USA; Department of Global Health, University of Washington, Box 359931, Seattle, WA 98195, USA. Electronic address: david.sherman@seattlebiomed.org.
authors: David R
authors: DR
authors: Sherman
authors: Benaroya Research Institute at Virginia Mason, 1201 9th Ave., Seattle, WA, 98101, USA. Electronic address: bkwok@benaroyaresearch.org.
authors: William W
authors: WW
authors: Kwok
authors: Benaroya Research Institute at Virginia Mason, 1201 9th Ave., Seattle, WA, 98101, USA. Electronic address: jyang@benaroyaresearch.org.
authors: Junbao
authors: J
authors: Yang
authors: Division of Infectious Diseases, St. John's Research Institute, St. John's National Academy of Health Sciences, Sarjapur Road, Koramangala 2 Block, Bangaluru, Karnataka 560034, India. Electronic address: Johnkennet@gmail.com.
authors: John
authors: J
authors: Kenneth
authors: Division of Infectious Diseases, St. John's Research Institute, St. John's National Academy of Health Sciences, Sarjapur Road, Koramangala 2 Block, Bangaluru, Karnataka 560034, India. Electronic address: mail2fazil@gmail.com.
authors: Syed F
authors: SF
authors: Ahamed
authors: ICGEB-Emory Vaccine Center, International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India; Emory Vaccine Center, 1510 Clifton Road, Atlanta, GA 30329, USA. Electronic address: chandeleanmol@gmail.com.
authors: Anmol
authors: A
authors: Chandele
authors: ICGEB-Emory Vaccine Center, International Center for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India; Emory Vaccine Center, 1510 Clifton Road, Atlanta, GA 30329, USA; Department of Pediatrics, Emory University, 1760 Haygood Drive, Atlanta, GA 30322, USA. Electronic address: murali.kaja@emory.edu.
authors: Kaja
authors: K
authors: Murali-Krishna
authors: Department of Medicine, Division of Infectious Diseases, University of Washington, Box 358061, Seattle, WA 98195, USA; Department of Global Health, University of Washington, Box 359931, Seattle, WA 98195, USA; Benaroya Research Institute at Virginia Mason, 1201 9th Ave., Seattle, WA, 98101, USA; Department of Laboratory Medicine, University of Washington, Box 358070, Seattle, WA 98195, USA; Vaccine and Infectious Diseases Division, Fred Hutchinson Cancer Research Center, 1100 Eastlake Ave. East, Seattle, WA 98109, USA. Electronic address: viralimm@u.washington.edu.
authors: David M
authors: DM
authors: Koelle
mesh_terms: D000328:Adult; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000382:microbiology; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D003429:Cross Reactions; D016207:Cytokines / Q000276:immunology; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D055985:Latent Tuberculosis / Q000276:immunology* / Q000382:microbiology; D008213:Lymphocyte Activation; D008875:Middle Aged; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology*; D016366:Open Reading Frames; D040901:Proteomics / Q000379:methods*; D032581:Tuberculosis Vaccines / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D016207:Cytokines; D018984:Epitopes, T-Lymphocyte; D032581:Tuberculosis Vaccines
keywords: Antigen; CD137; CD4; Malate synthase; T-cell; Tetramer; Tuberculosis