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A simple assay to quantify mycobacterial lipid antigen-specific T cell receptors in human tissues and blood.
- Source :
-
PLoS neglected tropical diseases [PLoS Negl Trop Dis] 2021 Dec 16; Vol. 15 (12), pp. e0010018. Date of Electronic Publication: 2021 Dec 16 (Print Publication: 2021). - Publication Year :
- 2021
-
Abstract
- T cell receptors (TCRs) encode the history of antigenic challenge within an individual and have the potential to serve as molecular markers of infection. In addition to peptide antigens bound to highly polymorphic MHC molecules, T cells have also evolved to recognize bacterial lipids when bound to non-polymorphic CD1 molecules. One such subset, germline-encoded, mycolyl lipid-reactive (GEM) T cells, recognizes mycobacterial cell wall lipids and expresses a conserved TCR-ɑ chain that is shared among genetically unrelated individuals. We developed a quantitative PCR assay to determine expression of the GEM TCR-ɑ nucleotide sequence in human tissues and blood. This assay was validated on plasmids and T cell lines. We tested blood samples from South African subjects with or without tuberculin reactivity or with active tuberculosis disease. We were able to detect GEM TCR-ɑ above the limit of detection in 92% of donors but found no difference in GEM TCR-ɑ expression among the three groups after normalizing for total TCR-ɑ expression. In a cohort of leprosy patients from Nepal, we successfully detected GEM TCR-ɑ in 100% of skin biopsies with histologically confirmed tuberculoid and lepromatous leprosy. Thus, GEM T cells constitute part of the T cell repertoire in the skin. However, GEM TCR-ɑ expression was not different between leprosy patients and control subjects after normalization. Further, these results reveal the feasibility of developing a simple, field deployable molecular diagnostic based on mycobacterial lipid antigen-specific TCR sequences that are readily detectable in human tissues and blood independent of genetic background.<br />Competing Interests: The authors have declared that no competing interests exist.
- Subjects :
- Antigens, CD1 genetics
Antigens, CD1 immunology
Cell Wall genetics
Cell Wall immunology
Cohort Studies
Humans
Leprosy blood
Leprosy immunology
Leprosy microbiology
Mycobacterium genetics
Mycobacterium isolation & purification
Nepal
Polymerase Chain Reaction
Receptors, Antigen, T-Cell, alpha-beta blood
Receptors, Antigen, T-Cell, alpha-beta genetics
South Africa
T-Lymphocytes immunology
T-Lymphocytes microbiology
Tuberculosis blood
Tuberculosis immunology
Tuberculosis microbiology
Leprosy diagnosis
Lipids immunology
Molecular Diagnostic Techniques methods
Mycobacterium immunology
Receptors, Antigen, T-Cell, alpha-beta immunology
Tuberculosis diagnosis
Subjects
Details
- Language :
- English
- ISSN :
- 1935-2735
- Volume :
- 15
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- PLoS neglected tropical diseases
- Publication Type :
- Academic Journal
- Accession number :
- 34914694
- Full Text :
- https://doi.org/10.1371/journal.pntd.0010018