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Blood, Vol. 106, Issue 3, 779-786, August 1, 2005
The tumor suppressor TSLC1/NECL-2 triggers NK-cell and CD8+ T-cell responses through the cell-surface receptor CRTAM
Blood Boles et al.
106: 779
Supplemental materials for: Boles et al, Vol 106, Issue 3, 779-786
Files in this Data Supplement:
- Figure S1. Multiple sequence alignment of a portion of the membrane distal Ig-like domains of the Nectin and Necl molecules and their receptors (JPG, 109 KB)
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Shading indicates the reliability of the alignment.
- Figure S2 CRTAM and Necl-2 expression and mAb binding on transfected cell lines (JPG, 47 KB)
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(A) Expression of CRTAM and Necl-2 on Daudi- and P815-transfected cells detected with the anti-FLAG mAb M2 (solid histograms) and compared with control mouse IgG1 (open histograms). (B) Cr24.1 stains CRTAM-Daudi (bottom panel), but does not recognize mock-transfected Daudi (top panel).
- Figure S3. CRTAM ligand is expressed on various epithelial cell lines and on a subset of B cells in the tonsil (JPG, 20 KB)
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(A) Flow cytometric analysis of epithelial (MZ-2, HeLa, HEK293, A549), neuroblastoma (SKNS1), B- (Daudi), T- (Jurkat), NK (NK92), erythroleukemia (K562), and endothelial (Huvec and Huvec treated with TNF-α and IFN-γ) cell lines, as well as immature (imMoDC) and CD40L-activated (CD40L-mMoDC) monocyte-derived dendritic cells with soluble CRTAM-mouse Fc. (B) Tonsil cells were labeled with soluble CRTAM-mouse Fc and CD20 mAb. Approximately 10% of the B cells expressed CRTAM ligand.
- Figure S4. Detection of mouse CRTAM and Necl-2 with Necl-2-Fc and mouse CRTAM-Fc, respectively (JPG, 39 KB)
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(A) Necl-2 binds mouse CRTAM. Daudi cells were transiently transfected with mouse CRTAM cDNA (mCRTAM-Daudi) or a control plasmid (mock-Daudi). Binding was performed with Necl-2-hFc. (B) Mouse NK cells up-regulate CRTAM upon coculture with YAC lymphoma cells. CRTAM expression on NK1.1+ NK cells was evaluated by Necl-2-hFc binding before and after stimulation and compared with a control soluble molecule (Nkp46-hFc). (C) A fraction of NK cells purified ex vivo from mice that rejected tumor cells express CRTAM. Cells were purified from peritoneal lavage of mice injected with Necl-2/EL-4 and EL-4 and stained with Necl-2-hFc and control-hFc and counterstained with NK1.1 and CD3. Necl-2-hFc+ cells are CD3– (not shown).
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