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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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Tissue distribution of MAIT cells. (A) Flow cytometry analysis of naive mouse thymus, enriched thymus, spleen, lymph nodes (inguinal), liver, lung, and lamina propria, showing reactivity to MR1–5-OP-RU tetramer or MR1–Ac-6-FP tetramer. Plots depict lymphocytes with B220+ B cells excluded by electronic gating. B6 mice are depicted in the top group and BALB/c in the bottom group. Numbers indicate the percentage of MAIT cells (red gate) of total αβ T cells (black gate). (B) Scatter plots depict MAIT cells as a proportion of T lymphocytes in all tissues tested, gated as shown in A. Each symbol represents an individual mouse. Bars depict mean ± SEM, and data are derived from a minimum of three independent experiments for each strain, with a combined total of 6–12 mice per tissue.
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fig2: Tissue distribution of MAIT cells. (A) Flow cytometry analysis of naive mouse thymus, enriched thymus, spleen, lymph nodes (inguinal), liver, lung, and lamina propria, showing reactivity to MR1–5-OP-RU tetramer or MR1–Ac-6-FP tetramer. Plots depict lymphocytes with B220+ B cells excluded by electronic gating. B6 mice are depicted in the top group and BALB/c in the bottom group. Numbers indicate the percentage of MAIT cells (red gate) of total αβ T cells (black gate). (B) Scatter plots depict MAIT cells as a proportion of T lymphocytes in all tissues tested, gated as shown in A. Each symbol represents an individual mouse. Bars depict mean ± SEM, and data are derived from a minimum of three independent experiments for each strain, with a combined total of 6–12 mice per tissue.

Mentions: Having identified MAIT cells in blood from WT mice, we next examined these cells in other tissues of two commonly used mouse strains (B6 and BALB/c), including thymus, spleen, liver, lymph node (inguinal), lung, and lamina propria (Fig. 2, A and B). Negative controls included MR1–Ac-6-FP tetramer (Fig. 2 A) and MR1–5-OP-RU tetramer staining of tissues from MR1−/− mice (similar to Fig. 1 C and not depicted). The data indicate the percentage of MAIT cells of total αβTCR+ cells. MAIT cells were difficult to detect in whole mouse thymus (<0.1%), but after complement-mediated depletion of immature thymocytes using anti-CD24, a clear population of MAIT cells was observed, representing ∼0.3% of mature (CD24 depleted) αβTCR+ thymocytes. MAIT cells were identified in all other tissues, although their frequency varied considerably. In B6 mice, the highest percentage was detected in lung (mean 3.3%), followed by lamina propria (mean 0.7%), liver (mean 0.6%), peripheral lymph nodes (mean 0.2%), spleen (mean 0.08%), and thymus (mean 0.05%; Fig. 2 B). In BALB/c mice, MAIT cells were generally less frequent as a percentage of total αβTCR+ cells, although a similar distribution was observed. In this strain, the liver housed the highest frequency of MAIT cells (mean 0.4%), followed by lung (mean 0.2%), spleen (mean 0.04%), thymus (mean 0.02%), and lymph nodes (mean 0.01%; Fig. 2, A and B). BALB/c lamina propria was not tested. In BALB/c thymus, MAIT cells were only clearly detectable after depletion of immature thymocytes with anti-CD24 antibody (Fig. 2 A). We were unable to detect a clear population of MAIT cells within the intraepithelial compartment of the gastrointestinal tract (not depicted). Thus, in mice, MAIT cells appear to be distributed in the same tissues as conventional T cells, although their relative frequency appears to be differentially regulated compared with control conventional T cells.


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)

Tissue distribution of MAIT cells. (A) Flow cytometry analysis of naive mouse thymus, enriched thymus, spleen, lymph nodes (inguinal), liver, lung, and lamina propria, showing reactivity to MR1–5-OP-RU tetramer or MR1–Ac-6-FP tetramer. Plots depict lymphocytes with B220+ B cells excluded by electronic gating. B6 mice are depicted in the top group and BALB/c in the bottom group. Numbers indicate the percentage of MAIT cells (red gate) of total αβ T cells (black gate). (B) Scatter plots depict MAIT cells as a proportion of T lymphocytes in all tissues tested, gated as shown in A. Each symbol represents an individual mouse. Bars depict mean ± SEM, and data are derived from a minimum of three independent experiments for each strain, with a combined total of 6–12 mice per tissue.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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

fig2: Tissue distribution of MAIT cells. (A) Flow cytometry analysis of naive mouse thymus, enriched thymus, spleen, lymph nodes (inguinal), liver, lung, and lamina propria, showing reactivity to MR1–5-OP-RU tetramer or MR1–Ac-6-FP tetramer. Plots depict lymphocytes with B220+ B cells excluded by electronic gating. B6 mice are depicted in the top group and BALB/c in the bottom group. Numbers indicate the percentage of MAIT cells (red gate) of total αβ T cells (black gate). (B) Scatter plots depict MAIT cells as a proportion of T lymphocytes in all tissues tested, gated as shown in A. Each symbol represents an individual mouse. Bars depict mean ± SEM, and data are derived from a minimum of three independent experiments for each strain, with a combined total of 6–12 mice per tissue.
Mentions: Having identified MAIT cells in blood from WT mice, we next examined these cells in other tissues of two commonly used mouse strains (B6 and BALB/c), including thymus, spleen, liver, lymph node (inguinal), lung, and lamina propria (Fig. 2, A and B). Negative controls included MR1–Ac-6-FP tetramer (Fig. 2 A) and MR1–5-OP-RU tetramer staining of tissues from MR1−/− mice (similar to Fig. 1 C and not depicted). The data indicate the percentage of MAIT cells of total αβTCR+ cells. MAIT cells were difficult to detect in whole mouse thymus (<0.1%), but after complement-mediated depletion of immature thymocytes using anti-CD24, a clear population of MAIT cells was observed, representing ∼0.3% of mature (CD24 depleted) αβTCR+ thymocytes. MAIT cells were identified in all other tissues, although their frequency varied considerably. In B6 mice, the highest percentage was detected in lung (mean 3.3%), followed by lamina propria (mean 0.7%), liver (mean 0.6%), peripheral lymph nodes (mean 0.2%), spleen (mean 0.08%), and thymus (mean 0.05%; Fig. 2 B). In BALB/c mice, MAIT cells were generally less frequent as a percentage of total αβTCR+ cells, although a similar distribution was observed. In this strain, the liver housed the highest frequency of MAIT cells (mean 0.4%), followed by lung (mean 0.2%), spleen (mean 0.04%), thymus (mean 0.02%), and lymph nodes (mean 0.01%; Fig. 2, A and B). BALB/c lamina propria was not tested. In BALB/c thymus, MAIT cells were only clearly detectable after depletion of immature thymocytes with anti-CD24 antibody (Fig. 2 A). We were unable to detect a clear population of MAIT cells within the intraepithelial compartment of the gastrointestinal tract (not depicted). Thus, in mice, MAIT cells appear to be distributed in the same tissues as conventional T cells, although their relative frequency appears to be differentially regulated compared with control conventional T cells.

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