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Blood, Vol. 111, Issue 6, 3108-3115, March 15, 2008
Recruitment of circulating NK cells through decidual tissues: a possible mechanism controlling NK cell accumulation in the uterus during early pregnancy
Blood Carlino et al.
111: 3108
Supplemental materials for: Carlino et al
Files in this Data Supplement:
- Table S1. Chemokine receptors and integrins expression on pbNK cells (PDF, 43 KB)
- Figure S1. Characterization of decidual endothelial cell primary cultures (JPG, 23.6 KB)
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DEC primary cultures were stained with Ab anti-vWF (A), anti-CD31 ULEX (B), anti-CD105 (C) and anti-VE-cadherin (D) and then assayed by immunofluorescence and cytofluorimetric analysis. Black histograms represent negative controls. The purity of all DEC primary cultures used was more than 98%. One representative staining is shown.
- Figure S2. Characterization of decidual stromal cell primary cultures (JPG, 47.3 KB)
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ST cell primary cultures were stained with anti-vimentin (A), anti-CK8-18 (B), anti-vWF (C), anti-CD31 (D), anti-smooth muscle alpha actin (F) upon fixation and permeabilization or with anti-CD105 (E), anti-CD10 (G) and anti-CD13 (H) and then assayed by immunofluorescence and cytofluorimetric analysis. Black histograms represent negative controls. The purity of all ST cell primary cultures used was more than 98%. One representative staining is shown.
- Figure S3. Progesterone receptor expression on DEC and ST cell primary cultures (JPG, 77.9 KB)
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Immunohistochemical analysis on DEC (panels A and B) and ST (panels C and D) cells pre-fixed with 10% buffered formalin and then stained with anti-progesterone receptor Ab (clone 1A6, Novocastra, UK) (panels A and C) or control Ab (panels B and D) for 1 hour at room temperature. The bound Ab was revealed using the labeled streptavidin biotin plus horseradish peroxidase kit and diaminobenzidine as substrate (Dako, Milano, Italy). Sections were counterstained with hematoxylin. One representative staining experiment of three performed is shown. Slides were evaluated using a Leica DM3000 optical microscope (Leica, Germany) and images were collected using Leica DFC320 digital camera at 400× magnification.
- Figure S4. Oestrogen enhances chemokine mRNA expression in decidual ST cells (JPG, 24.6 KB)
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mRNA isolated from primary cultures of decidual ST cells grown with or without oestrogen (1 nM) were analyzed for the expression of CXCL10/IP-10, CX3CL1/Fractalkine, CCL2/MCP-1 and CXCL12/SDF-1 by real time quantitative PCR assay as described in figure 1 panel D. Similar results were observed in two independent experiments. *, P < 0.05, as evaluated by performing statistical analysis between oestrogen versus non-oestrogen grown cells using Student’s t test.
- Figure S5. Chemokine receptor expression on pbNK and dNK cells of women in the first trimester of pregnancy (JPG, 50.7 KB)
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The chemokine receptor expression on NK cells freshly isolated from peripheral blood (PB) or decidual tissue (Decidua), obtained from the same women in the first trimester of pregnancy, was evaluated by performing a three color immunofluorescence and cytofluorimetric analysis as described in the material and method section. The dot plots shown indicate the expression of chemokine receptors on gated CD56+CD3− NK cells. Control represent staining with FITC-conjugated GAM or GARB Abs. Data are representative of four independent experiments.
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