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Topical application of a vitamin D3 analogue and corticosteroid to psoriasis plaques decreases skin infiltration of TH17 cells and their ex vivo expansion 

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Presentation on theme: "Topical application of a vitamin D3 analogue and corticosteroid to psoriasis plaques decreases skin infiltration of TH17 cells and their ex vivo expansion "— Presentation transcript:

1 Topical application of a vitamin D3 analogue and corticosteroid to psoriasis plaques decreases skin infiltration of TH17 cells and their ex vivo expansion  Toshiharu Fujiyama, MD, PhD, Taisuke Ito, MD, PhD, Takatsune Umayahara, MD, Shigeki Ikeya, MD, Kazuki Tatsuno, MD, Atsuko Funakoshi, PhD, Hideo Hashizume, MD, PhD, Yoshiki Tokura, MD, PhD  Journal of Allergy and Clinical Immunology  Volume 138, Issue 2, Pages e5 (August 2016) DOI: /j.jaci Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

2 Fig 1 Clinical pictures of psoriatic plaques before and 2 weeks after topical treatment in 5 patients. Cntrl, Control. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

3 Fig 2 Histopathologic changes in skin biopsy specimens from psoriatic lesions after a 2-week topical treatment. A, Hematoxylin and eosin staining of specimens. Black bar = 250 μm. B, Thickness of the epidermis and stratum corneum and percentage of parakeratosis length analyzed by using digital image analyzer software. *P < .05. Cntrl, Control. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

4 Fig 3 Numbers of CD3+ and IL-17+ cells analyzed by using digital image analyzer software. A, Representative immunohistochemical staining for CD3 (case 1) and immunofluorescence staining for IL-17A (case 2). Black bar = 250 μm; white bar, 100 μm. B, Positive cell numbers in the 10 patients and means ± SDs. *P < .05. Cntrl, Control. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

5 Fig 4 A–C, Number and frequency of CD4 and CD8 cells among expanded cells. T cells were expanded with anti-CD3/CD28 mAbs and IL-2 from biopsied skin topically treated with CB, Cal, or Bet or left untreated. *P < .05. Cntrl, Control. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

6 Fig 5 A–H, Number and frequency of expanded cells. T cells were expanded with anti-CD3/CD28 mAbs and IL-2 from biopsied skin topically treated with CB, Cal, or Bet or left untreated. They were analyzed in the surface phenotype and intracellular cytokines by using flow cytometric analysis. *P < .05. Cntrl, Control. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

7 Fig 6 Cytokine production by T cells from control (Cntrl) skin in response to anti-CD3 antibody stimulation under the conditions of 100 nmol/L Cal, 1 μmol/L Bet, or both (Cal+Bet). *P < .05. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

8 Fig 7 A–D, Total cell number and frequency of live cells and frequency of Annexin V+ cells among 7AAD− cells. Cells were cultured for 96 hours under conditions of 1 μmol/L Bet, 100 nmol/L Cal, or both (Cal+Bet). *P < .05. Cntrl, Control. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

9 Fig 8 TH17-related gene expression in lesional skin and expanded T cells. A, Relative mRNA expression for IL17A, CCL20, and IL23A in skin samples from 3 representative patients (cases 6, 8, and 10) topically treated with CB, Cal, or Bet or left untreated (Cntrl). Whole skin samples were homogenized to extract mRNA and subjected to quantitative RT-PCR. B, Relative mRNA expression for RORC and RORα in expanded T cells from nontreated or Cal-treated skin specimens of 5 patients (cases 1-5). Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

10 Fig E1 SUM score, which includes the severity (0- to 4-point scale) of erythema, induration, and scaling of the 10 cases. Cntrl, Control. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

11 Fig E2 Cytokine profiles of CD4+ (upper panels) and CD4+CD45RO+ (memory helper T cells; lower panels) cells before and after expansion. CD4+ T cells were isolated from PBMCs from 5 healthy donors and expanded with IL-2 and anti-CD3/CD28 antibodies for 2 weeks. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

12 Fig E3 Cytokine concentration of culture supernatants. T cells were expanded with anti-CD3/CD28 mAbs and IL-2 from biopsied skin topically treated CB, Cal, or Bet or left untreated. Cytokine production by T cells in response to anti-CD3 antibody stimulation was assessed by means of Cytokine Bead Array measurement of cytokines in the culture supernatants. *P < .05. Cntrl, Control. Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

13 Fig E4 TH1- and TH2-related gene expression in lesional skin. Relative mRNA expression for IL4 and IFN-γ in skin samples from 3 representative patients (cases 6, 8, and 10) topically treated with CB, Cal, or Bet or left untreated (Cntrl). Journal of Allergy and Clinical Immunology  , e5DOI: ( /j.jaci ) Copyright © 2016 American Academy of Allergy, Asthma & Immunology Terms and Conditions

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