학술논문

Identification of novel Mycobacterium tuberculosis CD4 T-cell antigens via high throughput proteome screening
Document Type
article
Source
Tuberculosis. 95(3)
Subject
Medical Microbiology
Biomedical and Clinical Sciences
Immunology
HIV/AIDS
Immunization
Clinical Research
Orphan Drug
Biotechnology
Prevention
Biodefense
Vaccine Related
Infectious Diseases
Tuberculosis
Rare Diseases
Infection
Good Health and Well Being
Adult
Antigens
Bacterial
Bacterial Proteins
CD4-Positive T-Lymphocytes
Cell Proliferation
Cell Separation
Cells
Cultured
Cross Reactions
Cytokines
Epitopes
T-Lymphocyte
High-Throughput Screening Assays
Humans
Latent Tuberculosis
Lymphocyte Activation
Middle Aged
Mycobacterium tuberculosis
Open Reading Frames
Proteomics
Tuberculosis Vaccines
CD4
T-cell
Antigen
CD137
Malate synthase
Tetramer
Medical and Health Sciences
Microbiology
Clinical sciences
Medical microbiology
Language
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.