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Identification of phenotypically and functionally heterogeneous mouse mucosal-associated invariant T cells using MR1 tetramers.

Rahimpour A, Koay HF, Enders A, Clanchy R, Eckle SB, Meehan B, Chen Z, Whittle B, Liu L, Fairlie DP, Goodnow CC, McCluskey J, Rossjohn J, Uldrich AP, Pellicci DG, Godfrey DI - J. Exp. Med. (2015)

Bottom Line: These cells include CD4(-)CD8(-), CD4(-)CD8(+), and CD4(+)CD8(-) subsets, and their frequency varies in a tissue- and strain-specific manner.Mouse MAIT cells have a CD44(hi)CD62L(lo) memory phenotype and produce high levels of IL-17A, whereas other cytokines, including IFN-γ, IL-4, IL-10, IL-13, and GM-CSF, are produced at low to moderate levels.These observations contrast with previous reports that MAIT cells from Vα19 TCR transgenic mice are PLZF(-) and express a naive CD44(lo) phenotype.

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Affiliation: Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity and Australian Research Council Centre of Excellence in Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria 3010, Australia Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity and Australian Research Council Centre of Excellence in Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria 3010, Australia.

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Phenotypic diversity within mouse MAIT cells. (A) Flow cytometry analysis of CD4 and CD8 coreceptor expression on MAIT cells from thymus, spleen, lymph nodes (inguinal), liver, and lung from B6 mice. (B) CD4/CD8 coreceptor–defined subsets of MAIT cells as a proportion of total MAIT cells in each tissue, gated as shown in A. Each symbol represents an individual mouse. Bar graphs depict mean ± SEM. Data are derived from a minimum of three independent experiments for each strain, with a combined total of 6–10 mice per tissue. (C) Phenotypic analysis of MAIT cells (thick black lines) from thymus, spleen, and lung compared with other (MR1–5-OP-RU tetramer− TCR-β+) T cells (shaded gray) depicted as histograms. Histograms for each marker, apart from CD218, show data concatenated from three separate mouse tissue samples from within one experiment and are representative of three similar experiments. Histograms for CD218 are representative of two experiments with four mice per experiment. (D) CD3+ and 5-OP-RU–reactive cells were stained for a panel of Vβ antibodies, and positive cells were plotted as a proportion of total MAIT cells. Bar graphs depict mean ± SEM from three independent experiments, each involving a pool of 10 mice.
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fig3: Phenotypic diversity within mouse MAIT cells. (A) Flow cytometry analysis of CD4 and CD8 coreceptor expression on MAIT cells from thymus, spleen, lymph nodes (inguinal), liver, and lung from B6 mice. (B) CD4/CD8 coreceptor–defined subsets of MAIT cells as a proportion of total MAIT cells in each tissue, gated as shown in A. Each symbol represents an individual mouse. Bar graphs depict mean ± SEM. Data are derived from a minimum of three independent experiments for each strain, with a combined total of 6–10 mice per tissue. (C) Phenotypic analysis of MAIT cells (thick black lines) from thymus, spleen, and lung compared with other (MR1–5-OP-RU tetramer− TCR-β+) T cells (shaded gray) depicted as histograms. Histograms for each marker, apart from CD218, show data concatenated from three separate mouse tissue samples from within one experiment and are representative of three similar experiments. Histograms for CD218 are representative of two experiments with four mice per experiment. (D) CD3+ and 5-OP-RU–reactive cells were stained for a panel of Vβ antibodies, and positive cells were plotted as a proportion of total MAIT cells. Bar graphs depict mean ± SEM from three independent experiments, each involving a pool of 10 mice.

Mentions: To investigate MAIT cell subsets and examine the activation and differentiation state of mouse MAIT cells, we colabeled MR1–5-OP-RU tetramer+ MAIT cells from B6 and BALB/c strains with antibodies specific for CD4 and CD8α (Fig. 3, A and B). In all tissues tested from B6 mice, the majority of MAIT cells were CD4−CD8−, although CD4+ and CD8+ subsets were also detected in lower proportions and with varied frequencies in a tissue-specific manner. For example, very few CD8+ MAIT cells (<5%) were detected in lymph nodes, whereas CD4+ MAIT cells were abundant (∼40%) in this tissue (Fig. 3, A and B), whereas CD4+ MAIT cells were virtually undetectable in lung. Furthermore, the expression level of CD4 and CD8 on MAIT cells were typically lower than on conventional T cells (Fig. 3 A and not depicted). CD8α+ MAIT cells in mice included a mix of both CD8α+β− and CD8α+β+ cells at roughly equal ratios, similar to human CD8+ MAIT cells (not depicted; Martin et al., 2009; Walker et al., 2012; Reantragoon et al., 2013). BALB/c MAIT cells also included the same three subsets based on CD4 and CD8 expression, although in this strain there were more CD8+ MAIT cells in thymus and a similar ratio of double-negative and CD8+ MAIT cells in spleen, lymph node, and liver (Fig. 3 B). Interestingly, similar to B6 mice, CD4+ MAIT cells were abundant (55%) in lymph nodes from BALB/c mice, suggesting that CD4+ MAIT cells preferentially reside in these organs.


Identification of phenotypically and functionally heterogeneous mouse mucosal-associated invariant T cells using MR1 tetramers.

Rahimpour A, Koay HF, Enders A, Clanchy R, Eckle SB, Meehan B, Chen Z, Whittle B, Liu L, Fairlie DP, Goodnow CC, McCluskey J, Rossjohn J, Uldrich AP, Pellicci DG, Godfrey DI - J. Exp. Med. (2015)

Phenotypic diversity within mouse MAIT cells. (A) Flow cytometry analysis of CD4 and CD8 coreceptor expression on MAIT cells from thymus, spleen, lymph nodes (inguinal), liver, and lung from B6 mice. (B) CD4/CD8 coreceptor–defined subsets of MAIT cells as a proportion of total MAIT cells in each tissue, gated as shown in A. Each symbol represents an individual mouse. Bar graphs depict mean ± SEM. Data are derived from a minimum of three independent experiments for each strain, with a combined total of 6–10 mice per tissue. (C) Phenotypic analysis of MAIT cells (thick black lines) from thymus, spleen, and lung compared with other (MR1–5-OP-RU tetramer− TCR-β+) T cells (shaded gray) depicted as histograms. Histograms for each marker, apart from CD218, show data concatenated from three separate mouse tissue samples from within one experiment and are representative of three similar experiments. Histograms for CD218 are representative of two experiments with four mice per experiment. (D) CD3+ and 5-OP-RU–reactive cells were stained for a panel of Vβ antibodies, and positive cells were plotted as a proportion of total MAIT cells. Bar graphs depict mean ± SEM from three independent experiments, each involving a pool of 10 mice.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4493408&req=5

fig3: Phenotypic diversity within mouse MAIT cells. (A) Flow cytometry analysis of CD4 and CD8 coreceptor expression on MAIT cells from thymus, spleen, lymph nodes (inguinal), liver, and lung from B6 mice. (B) CD4/CD8 coreceptor–defined subsets of MAIT cells as a proportion of total MAIT cells in each tissue, gated as shown in A. Each symbol represents an individual mouse. Bar graphs depict mean ± SEM. Data are derived from a minimum of three independent experiments for each strain, with a combined total of 6–10 mice per tissue. (C) Phenotypic analysis of MAIT cells (thick black lines) from thymus, spleen, and lung compared with other (MR1–5-OP-RU tetramer− TCR-β+) T cells (shaded gray) depicted as histograms. Histograms for each marker, apart from CD218, show data concatenated from three separate mouse tissue samples from within one experiment and are representative of three similar experiments. Histograms for CD218 are representative of two experiments with four mice per experiment. (D) CD3+ and 5-OP-RU–reactive cells were stained for a panel of Vβ antibodies, and positive cells were plotted as a proportion of total MAIT cells. Bar graphs depict mean ± SEM from three independent experiments, each involving a pool of 10 mice.
Mentions: To investigate MAIT cell subsets and examine the activation and differentiation state of mouse MAIT cells, we colabeled MR1–5-OP-RU tetramer+ MAIT cells from B6 and BALB/c strains with antibodies specific for CD4 and CD8α (Fig. 3, A and B). In all tissues tested from B6 mice, the majority of MAIT cells were CD4−CD8−, although CD4+ and CD8+ subsets were also detected in lower proportions and with varied frequencies in a tissue-specific manner. For example, very few CD8+ MAIT cells (<5%) were detected in lymph nodes, whereas CD4+ MAIT cells were abundant (∼40%) in this tissue (Fig. 3, A and B), whereas CD4+ MAIT cells were virtually undetectable in lung. Furthermore, the expression level of CD4 and CD8 on MAIT cells were typically lower than on conventional T cells (Fig. 3 A and not depicted). CD8α+ MAIT cells in mice included a mix of both CD8α+β− and CD8α+β+ cells at roughly equal ratios, similar to human CD8+ MAIT cells (not depicted; Martin et al., 2009; Walker et al., 2012; Reantragoon et al., 2013). BALB/c MAIT cells also included the same three subsets based on CD4 and CD8 expression, although in this strain there were more CD8+ MAIT cells in thymus and a similar ratio of double-negative and CD8+ MAIT cells in spleen, lymph node, and liver (Fig. 3 B). Interestingly, similar to B6 mice, CD4+ MAIT cells were abundant (55%) in lymph nodes from BALB/c mice, suggesting that CD4+ MAIT cells preferentially reside in these organs.

Bottom Line: These cells include CD4(-)CD8(-), CD4(-)CD8(+), and CD4(+)CD8(-) subsets, and their frequency varies in a tissue- and strain-specific manner.Mouse MAIT cells have a CD44(hi)CD62L(lo) memory phenotype and produce high levels of IL-17A, whereas other cytokines, including IFN-γ, IL-4, IL-10, IL-13, and GM-CSF, are produced at low to moderate levels.These observations contrast with previous reports that MAIT cells from Vα19 TCR transgenic mice are PLZF(-) and express a naive CD44(lo) phenotype.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity and Australian Research Council Centre of Excellence in Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria 3010, Australia Department of Microbiology and Immunology, Peter Doherty Institute for Infection and Immunity and Australian Research Council Centre of Excellence in Advanced Molecular Imaging, University of Melbourne, Parkville, Victoria 3010, Australia.

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Related in: MedlinePlus