NKG2D-L, ERp5, and ADAM10 expression in cHL. Twenty-five cHLs were studied. (A) Quantitative RT-PCR for MIC-A/B, ULBP1 to ULBP4, TG, ERp5, and ADAM10 was performed with the specific primers and probes on RNA extracted from the whole LN sections (8 μm). After subtracting the Ct value for RPLP0 from the Ct values of the target genes, results were expressed as ΔCt. Data are the mean ± SD from 15 NS samples (black columns) and 10 MC samples (gray columns). *P < .001 versus healthy LN (supplemental Figure 1A). (B) TG staining on NS (Bii) or MC (Biv) cHL sections (4 μm) One representative staining of 20 (11 NS and 9 MC). (Bi,iii) Negative control (CTR) on NS or MC serial sections, respectively. (C) Quantitative RT-PCR of stromal (S) or parenchymal (P) microdissected areas of serial sections (n = 20 cHL: 11 NS and 9 MC) that underwent LCM on the basis of the localization of TG. Quantitative RT-PCR for MIC-A/B, ULBP2, ULBP3, TG, ERp5, and ADAM10 was performed as in panel A, normalized for RPLP0, and results calculated as ΔCt. Data are the mean ± SD from 10 S and 10 P areas cut from 20 cHL. *P < .001 S versus parenchymal. (D-E) Immunostaining for MIC-A (Di), ULBP3 (Dii), ERp5 (Diii,Eiii) ADAM10 (Div,Eiv), and CD30 (Dii) on LN sections from a representative NS (D) or MC (E) cHL of 11 NS and 9 MC analyzed. Arrows indicate positive areas. (Ei) Negative control. Arrows in panel E indicates RS cells. The slides were analyzed under an IX70 microscope (Olympus Biosystem) equipped with a camera (Camedia 4040Zoom, Olympus; original magnification ×200 with a 20×/0.50 NA objective). Black bar represents 10 μm.
