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Dermal Vγ4+ γδ T Cells Possess a Migratory Potency to the Draining Lymph Nodes and Modulate CD8+ T-Cell Activity through TNF-α Production  Satoshi Nakamizo,

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Presentation on theme: "Dermal Vγ4+ γδ T Cells Possess a Migratory Potency to the Draining Lymph Nodes and Modulate CD8+ T-Cell Activity through TNF-α Production  Satoshi Nakamizo,"— Presentation transcript:

1 Dermal Vγ4+ γδ T Cells Possess a Migratory Potency to the Draining Lymph Nodes and Modulate CD8+ T-Cell Activity through TNF-α Production  Satoshi Nakamizo, Gyohei Egawa, Michio Tomura, Shunsuke Sakai, Soken Tsuchiya, Akihiko Kitoh, Tetsuya Honda, Atsushi Otsuka, Saeko Nakajima, Teruki Dainichi, Hideaki Tanizaki, Masao Mitsuyama, Yukihiko Sugimoto, Kazuhiro Kawai, Yasunobu Yoshikai, Yoshiki Miyachi, Kenji Kabashima  Journal of Investigative Dermatology  Volume 135, Issue 4, Pages (April 2015) DOI: /jid Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Migration of gamma delta T cells (γδ T cells) from the skin to the draining lymph nodes (LNs). (a) Flow cytometry of the skin of Kaede-tg mice before (left) and immediately after (right) the violet light exposure. (b) Flow cytometry of Kaede-red+ cells in the draining popliteal LNs 24 hours after the photoconversion of the footpad. Cells were gated on CD11c+ (left) or γδ TCR+ (right) cells. (c) The number of Kaede-red+ CD11c+ (left) or γδ TCR+ cells (right) in the draining LNs 3 days after the intradermal injection of BCG. The cells in the footpad were photoconverted 24 hours before the analysis. Data are representative of three experiments (n=3) and are presented as means±SD. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Gamma delta T cells (γδ T cells) migrate from the skin to the draining lymph nodes (LNs) in a Gi-independent manner. (a, b) Flow cytometry of CD11c+ (left) and γδ TCR+ (right) cells in the skin-draining LNs of WT (upper panel) and CCR7-deficient (lower panel) Kaede-tg mice 24 hours after the photoconversion of the skin. The % frequencies of Kaede-red+ cells are shown (b). (c) Flow cytometry of CCR7 expression on γδ T cells in the skin (left panel) and in the skin-draining LN 24 hours after the photoconversion (right panel). (d, e) Flow cytometry of CD11c+ (left) and γδ TCR+ (right) cells in the draining LNs of Kaede-tg mice 24 hours after the photoconversion of the skin with pertussis toxin- or phosphate-buffered saline treatment. The % frequencies of Kaede-red+ cells were shown (e). Data are representative of three experiments (n=3) and are presented as means±SD. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Skin-derived gamma delta T cells (γδ T cells) are Vγ4+ dermal γδ T cells. (a) Flow cytometric analysis of Kaede-red+ γδ T cells in the draining LNs 24 hours after the photoconversion of the skin. The % frequencies of Vγ4+ cells are shown in the right panel. (b) Flow cytometric analysis of Vγ4+ cells in the skin-draining LNs of Kaede-tg mice 24 hours after photoconversion of the skin cells. (c) Flow cytometric analysis of Kaede-red+ Vγ4+ cells in the dermis. (d) Flow cytometric analysis of Vγ4+ cells in the skin-draining LNs. (e) Flow cytometric analysis of Kaede-red+ Vγ4+ cells in the skin-draining LNs 24 hours after the photoconversion of the skin. (f) The % frequency of CCR6+CD103+ cells among Vγ4+ cells in the dermis, LN, and spleen. Data are representative of three experiments (n=3) and are presented as means±SD. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Vγ4+ cells enhance the intranodal expansion of CD8+ T cells against bacille Calmette–Guérin (BCG). (a) Relative amount of Il17a and Tnfa mRNA expression in each subset of intranodal Vγ4+ cells. For each subset of Vγ4+ cells, equal amounts of total RNA were pooled from five mice. (b) Flow cytometric analysis of CTV-labeled OT-I tg T cells from control (Ctrl)- or neutralizing anti-Vγ4 antibody–treated mice 6 days after injection of OVA-expressing BCG. The number of CTVlow cells is shown in the lower panel. Data are representative of three experiments (n=4) and are presented as means±SD. *P<0.05. ND, not detected. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Vγ4+ gamma delta T cells (γδ T cells) stimulate antigen-specific CD8+ T-cell differentiation by enhancement of dendritic cell (DC) functions. (a) Immunohistochemical staining of the skin-draining LNs 24 hours after the photoconversion of the skin. B220+ (white) and γδ TCR+ (green) cells are shown. Right panel shows the higher-magnification view of the boxed area in the left panel. Red signals represent Kaede-red. Arrowheads in the right panel indicate Kaede-red+ γδ T cells (right). Scale bars=100 μm (left) and 50 μm (right). (b, c) The number of CTVlow (as an indication of cell proliferation) OT-I tg T cells (b) and IFN-γ-producing cells (c). CTV-labeled OT-I tg T cells were cocultured with CD11c+ DCs from OVA-expressing BCG–sensitized mice treated with control (Ctrl) or neutralizing anti-Vγ4 antibody. (d) The protein levels of IFN-γ and IL-12p40 in the coculture supernatant. Data are representative of three experiments (n=3∼4) and are presented as means±SD. *P<0.05. T, T-cell zone; B, B-cell zone. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 Vγ4+ gamma delta T cells (γδ T cells) stimulate bone marrow–derived dendritic cells (BMDCs) to produce IL-12p40. (a) The % frequency of IL-12p40+ BMDCs cultured with or without CD4+ T cells or Vγ4+ cells. (b) The mean fluorescence intensity (MFI) of IL-12p40 expression in BMDCs cultured with or without Vγ4+ cells. (c) The % frequency of IL-12p40+ BMDCs cultured with or without Vγ4+ γδ T cells in the presence of isotype control (Ctrl), IL-17RFc, and anti-TNF-α antibodies. (d) The number of TNF-α+ CD11c+ DCs (open column) and TNF-α+ Vγ4+ cells (filled column) in the skin-draining LNs 3 days after treatment without (day 0) or with BCG (day 3). Data are representative of three experiments (n=3) and are presented as means±SD. *P<0.05. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2015 The Society for Investigative Dermatology, Inc Terms and Conditions

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