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Soluble Peptide Treatment Reverses CD8 T-Cell-Induced Disease in a Mouse Model of Spontaneous Tissue-Selective Autoimmunity  So Yeon Paek, Fumi Miyagawa,

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Presentation on theme: "Soluble Peptide Treatment Reverses CD8 T-Cell-Induced Disease in a Mouse Model of Spontaneous Tissue-Selective Autoimmunity  So Yeon Paek, Fumi Miyagawa,"— Presentation transcript:

1 Soluble Peptide Treatment Reverses CD8 T-Cell-Induced Disease in a Mouse Model of Spontaneous Tissue-Selective Autoimmunity  So Yeon Paek, Fumi Miyagawa, Hong Zhang, Jay T. Linton, Shelley B. Hoover, R Mark Simpson, Stephen I. Katz  Journal of Investigative Dermatology  Volume 132, Issue 3, Pages (March 2012) DOI: /jid Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Development of keratin-14-soluble chicken ovalbumin/OT-I (#17) double-transgenic (DTg) mouse ear on days 1–9 after birth. Hematoxylin and eosin stain (a) DTg mice developed normally until birth, after which the pinnae (external ear auricles) developed bullae at the dermal–epidermal junction associated with progressive epidermal necrosis and leukocyte infiltration over days 3–6, resulting in the loss of epidermis on the pinnal surface, and eventually the loss of external ears by day 10. The crown of the mouse is on the right and the ventral aspect on the left (bars=100μm). (b) DTg mice ears displayed at higher resolution from mouse #17 at day 3 (d3; left and middle panels), and mouse #5 at d4 (right panel). The changes include evolving bullae at the dermal–epidermal junction associated with epidermal necrosis and leukocyte infiltration (bars=25μm). Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 OVA mRNA expression levels in different tissues from C57BL/6 and K14-sOVA (#5 and #17) adult single Tg mice. The OVA Tg mRNA expression levels were quantified by real-time PCR with mGAPDH as the housekeeping gene. The average of two mice from each group is shown. K14-sOVA, keratin-14-soluble chicken ovalbumin; OVA, ovalbumin; Tg, transgene. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Inflammatory infiltrates in K14-sOVA/OT-I (#5 and #17) mice ears. (a) Few T cells in the dermis of OVA-peptide-treated DTg mice, compared with more florid dermal infiltration of CD8+ T cells (red; 4,6-diamidino-2-phenylindole blue) observed in d1–3 DTg control mice (epidermis on right; original magnification × 40). (b) FACS of cell suspensions (liberase-digested) from day 5 DTg pup ears, and (c) histology of DTg mice at embryonic day 19 (day E19). Arrows represent the zone of epidermal–keratinocyte programmed cell death; open space at left arrow is precursor to external auditory canal (bars=50μm). (d) FACS data of spleens of day E18 DTg mice with Vα2/Vβ5-positive TCR (range 11.4–24.7%) and CD8+ T cells (range 10.8–23.9%). (e) Histology of ears of CD-8-depleted DTg mice is normal (bars=100μm). APC, allophycocyanin; DTg, double transgenic; K14-sOVA, keratin-14-soluble chicken ovalbumin; OVA, ovalbumin; PE, phycoerythrin. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 Marked reduction of OT-I cell numbers at earlier time points and downregulation of the CD8 co-receptor following peptide treatment. (a) FACS analysis of spleens of untreated and peptide-treated K14-sOVA/OT-I (#5 and #17) pups harvested on days E18, 3, 5, and 9 and stained for Vα2/Vβ5 (OT-I cells). (b) Spleens harvested and pooled from untreated K14-sOVA/OT-I (#5) and peptide-treated K14-sOVA/OT-I (#5 and #17) mice on days 3, 5, and 9 were stained for CD8. Mean fluorescence intensity of CD8 in OT-I cells was lower at all time points in peptide-treated mice compared with untreated mice. K14-sOVA, keratin-14-soluble chicken ovalbumin; PE, phycoerythrin. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 Activation markers on OT-I cells in vivo. OT-I cells of untreated and peptide-treated K14-sOVA/OT-I DTg pup spleens were stained for the surface markers CD25, CD44, CD62L, and CD69 on days 3, 5, and 9 after birth. K14-sOVA/OT-I pups treated with SIINFEKL peptide demonstrated an activated phenotype on postnatal day 3, but were CD25low CD44high CD62Lhigh CD69low by days 5 and 9. Legend: red, sOVA/OT-I (#5) untreated; blue, sOVA/OT-I (#5) peptide treated; green, sOVA/OT-I (#17) peptide treated. APC, allophycocyanin; DTg, double transgenic; K14-sOVA, keratin-14-soluble chicken ovalbumin. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

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