1. Role of CCL17 in the Generation of Cutaneous Inflammatory Reactions in Hu-PBMC- SCID Mice Grafted with Human Skin 
Jules Gilet, Ying Chang, Cécile Chenivesse, Benjamin Legendre, Han Vorng, Catherine Duez, Benoît Wallaert, Henri Porte, Stéphanie Senechal, Anne Tsicopoulos 
Journal of Investigative Dermatology 
Volume 129, Issue 4, Pages (April 2009)
DOI: /jid
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
Kinetics and distribution of intradermally injected human 125I-CCL I-CCL17 was injected intradermally into one skin graft (homolateral) and the control diluent into the contralateral skin graft of each mouse. Radioactivity was evaluated 15, 60, and 120minutes after injection in skin, skin draining homolateral and contralateral LN, kidney, mesenteric LN, spleen, liver, and blood. Tissues were weighed and counted on a γ-counter. The specific radioactivity associated with each organ was expressed per unit wet weight as the mean c.p.m.±SEM for n=4–6 per time point. *P<0.05.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Kinetics of cell recruitment in draining LN. Mice were reconstituted i.p. with autologous PBMC and CCL17 or the diluent were immediately injected into human skin xenografts. (a) Kinetics of T-cell recruitment in LN draining CCL17- versus diluent-injected sites, draining diluent/diluent-injected mice, or draining CCL17/anti-CCL17-injected sites. Cryostat sections of the draining LN were immunostained with monoclonal antibodies against CD45+ leukocytes, CD4+ T cells, CD45RO+ memory T cells. Results are expressed as the mean number±SEM of positive cells per lymph node section for n=6–8 mice per time point. *P<0.05. **P<0.01. (b) Kinetics of CCR4+ cell and DC recruitment in LN draining CCL17- versus diluent-injected sites, draining diluent/diluent-injected mice, or draining CCL17/anti-CCL17-injected sites. Immature DC were assessed using the CD1a antibody, mature DC using the DC-LAMP antibody. Results are expressed as above.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Human cell cutaneous recruitment in skin xenografts from humanized SCID mice 24hours after i.d. CCL17 versus diluent injections. (a) Characterization of the T-cell infiltrate. CCL17 was injected into human skin xenografts immediately after i.p. reconstitution with autologous PBMC. Cryostat sections of human skin biopsies were immunostained with monoclonal antibodies against CD45+ leukocytes, CD4+ and CD8+ T-cell subpopulations, CD45RO+ memory, CD45RA+ naïve, and CD25+ subpopulations. Results are expressed as number of positive cells per mm2 for n=8 mice. Mean values are represented by the solid bars. (b) Infiltration of BB1+ basophils, of murine major basic protein mMBP+ eosinophils, and of CD68+ monocytes/macrophages. Results are expressed as above.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Phenotypic characterization of the generated Th-1 and Th-2 subsets. (a) Th-1 and Th-2 subsets were stained with an annexin V/propidium iodide kit and analyzed by flow cytometry. Dot plots show the percentage of apoptotic cells (annexin V+/PI−) and the percentage of dead cells (annexin V+/PI+). (b) Th-1 and Th-2 cells were stained using a cytokine secretion assay kit. Histogram plots of IL-4 and IFN-γ secreting cells are overlaid (thick line) on isotype-matched control (thin line). (c) Histogram plots of Th-1 and Th-2 cells expressing CCR4 (thick line) overlaid on isotype-matched control (thin line). One representative experiment out of three is shown.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
Cutaneous versus lymph node recruitment of IL-4 and IFN-γ positive cells 24hours after i.d. CCL17 versus diluent injections in Th-1 or Th-2 reconstituted SCID mice. CD4, CCR4, IL-4, and IFN-γ positive cells were assessed by immunostaining on skin (a) or lymph node (b) cryostat sections. Results are shown as the mean number±SEM of positive cells per mm2 for skin, and per lymph node section for lymph nodes, for n=6 mice per group.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

7. Figure 6
Direct chemotactic effect of CCL17 on: (a) Human Th-2, Th-1, and CD45RA+ T cells. (b) Murine eosinophils, human basophils, and monocytes. Chemotaxis assays were performed using a Boyden chamber, with CCL17 at different doses and RPMI control. Results are expressed as the mean number of recruited cells per well for T cells and per field for the other cells.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

8. Figure 7
Microphotographs of immunohistochemistry on cryostat sections. (a) Skin draining LN 3hours after CCL17 intradermal injection immunostained with anti-CD45RO antibody. (b, c) Human skin graft 24hours after diluent (b) or CCL17 (c) injection, immunostained with anti-CD4 antibody. (d) IL-4 positive cells in a cryostat section of a human skin graft injected with CCL17 taken from a mouse reconstituted with polarized Th-2 cells. Arrows point out positive cells. Scale bar=50μm.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions