Validation of a CD1b tetramer assay for studies of human mycobacterial infection or vaccination

Layton, Erik D.; Yu, Krystle K. Q.; Smith, Malisa T.; Scriba, Thomas J.; Rosa, Stephen C. De; Seshadri, Chetan

doi:10.1016/j.jim.2018.04.004

Cited by 18 publications

(28 citation statements)

References 35 publications

(46 reference statements)

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“…Such positive findings have been seen in small (13,14) medium (15,16) and large studies (17,18). In contrast, this large-scale tetramer study, as well as smaller studies that did (36) or did not (11) use CD1 tetramers in South African cohorts, found no differences among highly exposed groups that differed in their IGRA status.…”

Section: Discussionmentioning

confidence: 56%

“…LDN5-like T cells were described as a TCR motif based on study of six T cell clones studied outside the context of TB (25). Polyclonal GEM T cells have been previously detected in small numbers of subjects (24,36), but their prevalence among humans, including TB patients, remained unknown. In 147 analyzable samples, we first gated CD1b-GMM tetramers + T cells to identify samples with 10 or more cells in the gate (n = 115, Supplementary Table 2).…”

Section: Subpopulations Of Cd1b-gmm Specific T Cellsmentioning

confidence: 99%

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CD1b Tetramers Broadly Detect T Cells That Correlate With Mycobacterial Exposure but Not Tuberculosis Disease State

et al. 2020

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The non-polymorphic nature of CD1 proteins creates a situation in which T cells with invariant T cell receptors (TCRs), like CD1d-specific NKT cells, are present in all humans. CD1b is an abundant protein on human dendritic cells that presents M. tuberculosis (Mtb) lipid antigens to T cells. Analysis of T cell clones suggested that semi-invariant TCRs exist in the CD1b system, but their prevalence in humans is not known. Here we used CD1b tetramers loaded with mycolic acid or glucose monomycolate to study polyclonal T cells from 150 Peruvian subjects. We found that CD1b tetramers loaded with mycolic acid or glucose monomycolate antigens stained TRAV1-2 + GEM T cells or TRBV4-1 + LDN5-like T cells in the majority of subjects tested, at rates ∼10-fold lower than NKT cells. Thus, GEM T cells and LDN5-like T cells are a normal part of the human immune system. Unlike prior studies measuring MHC-or CD1b-mediated activation, this large-scale tetramer study found no significant differences in rates of CD1b tetramer-mycobacterial lipid staining of T cells among subjects with Mtb exposure, latent Mtb infection or active tuberculosis (TB) disease. In all subjects, including "uninfected" subjects, CD1b tetramer + T cells expressed memory markers at high levels. However, among controls with lower mycobacterial antigen exposure in Boston, we found significantly lower frequencies of T cells staining with CD1b tetramers loaded with mycobacterial lipids. These data link CD1b-specific T cell detection to mycobacterial exposure, but not TB disease status, which potentially explains differences in outcomes among CD1-based clinical studies, which used control subjects with low Mtb exposure.

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Section: Discussionmentioning

confidence: 56%

Section: Subpopulations Of Cd1b-gmm Specific T Cellsmentioning

confidence: 99%

CD1b Tetramers Broadly Detect T Cells That Correlate With Mycobacterial Exposure but Not Tuberculosis Disease State

et al. 2020

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“…In vitro-derived T cell lines have been isolated from peripheral blood and lesion biopsies of individuals infected with M. leprae (15,18). GMM-and SGL-specific T cells have been detected at a higher frequency in individuals infected with M.tb than healthy individuals (16,19). Mycolipid-specific T cells have also been studied directly from the peripheral blood of individuals with active TB (20).…”

Section: Mycolipids As T Cell Antigensmentioning

confidence: 99%

“…It is unknown what factors are required for activation of naive mycolipid-specific T cells. Mycolipid-specific T cells are found at a higher frequency in individuals with documented exposure to mycobacteria, suggesting that prior antigen exposure drives activation and proliferation (16,19). There is evidence that immunization of cattle with GMM produces GMM-specific T cells with enhanced proliferative potential, also suggesting that exposure to antigen can lead to activation and enhanced recall responses (68).…”

Section: Mycolipid-specific T Cell Activation and Memory Phenotypementioning

confidence: 99%

T Cell Responses to Mycobacterial Glycolipids: On the Spectrum of “Innateness”

James

Seshadri

2020

Front. Immunol.

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Diseases due to mycobacteria, including tuberculosis, leprosy, and Buruli ulcer, rank among the top causes of death and disability worldwide. Animal studies have revealed the importance of T cells in controlling these infections. However, the specific antigens recognized by T cells that confer protective immunity and their associated functions remain to be definitively established. T cells that respond to mycobacterial peptide antigens exhibit classical features of adaptive immunity and have been well-studied in humans and animal models. Recently, innate-like T cells that recognize lipid and metabolite antigens have also been implicated. Specifically, T cells that recognize mycobacterial glycolipid antigens (mycolipids) have been shown to confer protection to tuberculosis in animal models and share some biological characteristics with adaptive and innate-like T cells. Here, we review the existing data suggesting that mycolipid-specific T cells exist on a spectrum of "innateness," which will influence how they can be leveraged to develop new diagnostics and vaccines for mycobacterial diseases.

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“…The gates proceeded from Live and CD3 + cells to CD14 - and CD19 - cells to single cells to a CCR7 “keeper gate” to lymphocytes by size gating. The CCR7 Keeper Gate is applied to eliminate events stained with dye aggregates, as these can interfere with downstream analysis and gating of rare cell populations (Layton et al, 2018). After this point, gates were drawn for γδ T cells, TRAV1-2, CD4 and CD8, and CD45RA and CCR7 independently for CD4 and CD8 T cells.…”

Section: Supplemental Figure Legendsmentioning

confidence: 99%

CD4 and CD8 co-receptors modulate functional avidity of CD1b-restricted T cells

James

Aguilar

et al. 2020

Preprint

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CD4 and CD8 co-receptors define distinct lineages of T cells restricted by major histocompatibility complex (MHC) Class II and I molecules, respectively. Co-receptors interact with the T cell receptor (TCR) at the surface of MHC-restricted T cells to facilitate antigen recognition, thymic selection, and functional differentiation. T cells also recognize lipid antigens presented by CD1 molecules, but the role that CD4 and CD8 play in lipid antigen recognition is unknown. We studied the effect of CD4 and CD8 on the avidity, activation, and function of T cells specific for two CD1b-presented mycobacterial lipid antigens, glucose monomycolate (GMM) and diacylated sulfoglycolipids (SGL). In a human cohort study using SGL-loaded CD1b tetramers, we discovered a hierarchy among SGL-specific T cells in which T cells expressing the CD4 or CD8 co-receptor stain with a higher tetramer mean fluorescence intensity (MFI) than CD4-CD8- T cells. To determine the role of the TCR co-receptor in lipid antigen recognition, we exogenously expressed GMM and SGL-specific TCRs in Jurkat or polyclonal T cells and quantified tetramer staining and activation thresholds. Transduced CD4+ primary T cells bound the lipid-loaded CD1b tetramer with a higher MFI than CD8+ primary T cells, and transduced CD8+ Jurkat cells bound the SGL-CD1b tetramer with higher MFI than CD4-CD8- Jurkat cells. The presence of either co-receptor also decreased the threshold for IFN-γ secretion. Further, co-receptor expression increased surface expression of CD3ε, suggesting a mechanism for increased tetramer binding and activation. Finally, we used single-cell sequencing to define the TCR repertoire and ex vivo functional profiles of SGL-specific T cells from individuals with M.tb disease. We found that CD8+ T cells specific for SGL express canonical markers associated with cytotoxic T lymphocytes, while CD4+ T cells could be classified as T regulatory or T follicular helper cells. Among SGL-specific T cells, only those expressing the CD4 co-receptor also expressed Ki67, suggesting that they were actively proliferating at the time of sample collection. Together, these data reveal that expression of CD4 and CD8 co-receptor modulates TCR avidity for lipid antigen, leading to functional diversity and differences in in vivo proliferation during M.tb disease.

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Validation of a CD1b tetramer assay for studies of human mycobacterial infection or vaccination

Cited by 18 publications

References 35 publications

CD1b Tetramers Broadly Detect T Cells That Correlate With Mycobacterial Exposure but Not Tuberculosis Disease State

CD1b Tetramers Broadly Detect T Cells That Correlate With Mycobacterial Exposure but Not Tuberculosis Disease State

T Cell Responses to Mycobacterial Glycolipids: On the Spectrum of “Innateness”

CD4 and CD8 co-receptors modulate functional avidity of CD1b-restricted T cells

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