CD158K/KIR3DL2 Transcript Detection in Lesional Skin of Patients with Erythroderma Is a Tool for the Diagnosis of Sézary Syndrome Nicolas Ortonne, Sabine.

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CD158K/KIR3DL2 Transcript Detection in Lesional Skin of Patients with Erythroderma Is a Tool for the Diagnosis of Sézary Syndrome Nicolas Ortonne, Sabine Le Gouvello, Hicham Mansour, Catherine Poillet, Nadine Martin, Marie-Hélène Delfau-Larue, Karen Leroy, Jean-Pierre Farcet, Martine Bagot, Armand Bensussan Journal of Investigative Dermatology Volume 128, Issue 2, Pages 465-472 (February 2008) DOI: 10.1038/sj.jid.5701013 Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Immunohistochemistry results for CD3 and CD158k/e (AZ158 mAb) in representative SS and EID skin samples. Histology, CD3 and AZ158 mAb stainings in cutaneous biopsies from (a, c, and e) the representative SS patient 8, and from (b, d, and f) a patient with erythrodermic psoriasis. (a and b) Hematoxylin and eosin-stained sections show in both samples a sub-epidermal band-like lymphocytic infiltrate associated with scattered epidermotropic lymphocytes. (c and d) CD3+ T cells are seen, but (c) a higher T-cell infiltrate is present in the SS sample, associated with Pautrier micro-abscesses (arrow). (e) Lymphocytes stained with AZ158 mAb are seen in both the dermis and the epidermis in the SS sample, and also (f) in a significant proportion of dermal lymphocytes (arrows) in the erythrodermic psoriasis cutaneous sample. Slides are shown at original magnification × 200 (a and b) and × 400 (c–f). Bar=0.1mm. Journal of Investigative Dermatology 2008 128, 465-472DOI: (10.1038/sj.jid.5701013) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Conventional RT–PCR amplification of CD158e/KIR3DL1 and CD158k/KIR3DL2 in erythrodermic skin samples. (a) Controls. In natural killer lymphocytes from a healthy donor, used as a positive control, both CD158e/KIR3DL1 and CD158k/KIR3DL2 transcripts are found (left), whereas in the Sézary cell line OGU, only CD158k/KIR3DL2 appears to be positive (middle). By contrast, in normal skin used as a negative control, both CD158e and CD158k RT–PCR are negative (right). (b) Skin biopsies from patients with erythroderma. In the skin sample from the EID group, neither CD158e/KIR3DL1 nor CD158k/KIR3DL2 mRNA expression is detected (left). By contrast, in the two SS samples (middle and right), a significant expression of CD158k/KIR3DL2 is found, with a bright distinctive band at the expected size, whereas CD158e/KIR3DL1 transcript are not detected. Journal of Investigative Dermatology 2008 128, 465-472DOI: (10.1038/sj.jid.5701013) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Quantitative RT–PCR amplification of CD3, CD158k/KIR3DL2 and T-plastin in erythrodermic skin samples. Activated peripheral blood mononuclear cells from a healthy control donor were used as a calibrator and arbitrarily set as a CD3, CD158e/KIR3DL1, CD158k/KIR3DL2 and T-plastin expression level of 1. The middle bar in each group represents the median of that group. (a) The median level of CD3 is higher in the SS group (P=0.0133) (left). CD158k/KIR3DL2 mRNA expression is highly overexpressed in SS versus EID skin samples (P<0.0001) (middle). By contrast, T-plastin transcript quantification is not discriminant between SS and EID (P=0.8421) (right). (b) Raw data (relative transcript expression × 100), showing a significant overexpression of CD158k/KIR3DL2 mRNA, but not T-plastin in SS (left) versus EID (right) skin samples. All data points represent the average of duplicate tests. Journal of Investigative Dermatology 2008 128, 465-472DOI: (10.1038/sj.jid.5701013) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 KIR3DL2 gene, RT–PCR products, and primers localization. Relative positions of the different cDNA segments (bottom) across the nine exons of the KIR3DL2 gene (top), amplified according to the different sets of primers used for conventional and quantitative RT–PCR. Journal of Investigative Dermatology 2008 128, 465-472DOI: (10.1038/sj.jid.5701013) Copyright © 2008 The Society for Investigative Dermatology, Inc Terms and Conditions