<p>PDF file - 223K, Supplemental Figure 4: Activation of pDC in response to MV-infected tumor cells is independent of MV replication in pDC and infection by CD46. (A) pDC were cultured with IL-3 alone, R848, IL-3/MV-eGFP (MV), IL-3/MV-eGFP with 10mg/ml of anti-CD46 or IL-3/UV-irradiated MV-eGFP (MV*) at MOI=1 during 18hrs. Expression of CD83, CD80 and CD86 by pDC was determined by flow cytometry. (B) pDC were cultured with IL-3, UV-irradiated M18, MV-infected M18 (M18MV) in presence or absence of 10mg/ml of anti-CD46 (Hycult biotech), or MV-infected M18 irradiated by UV before exposition to pDC (M18MV*). Expression of CD83, CD80 and CD86 by pDC (gate on BDCA-4+/HLA-DR+ cells) was determined by flow cytometry. (C) IFN-a production by pDC measured by ELISA. (D) MV-eGFP infection inhibition of M18 by 10mg/ml of anti-CD46 monoclonal antibody after 72hrs of culture.</p>
<p>PDF file - 115K, Supplemental figure 3: Infection of pDC by MV-eGFP or UV-irradiated MV-eGFP: pDC in presence of IL-3 or Meso13 cells were cultured alone (NI) or with MV-eGFP (MV) or UV-irradiated (312nm - 100kj/m�) MV-eGFP (MV*) at MOI=50 during 72hrs (upper panel) or during 2hrs and then cultured during 70hrs (lower panel). Fluorescence was analyzed by flow cytometry.</p>
Diagnosis of malignant pleural mesothelioma is a challenging issue. Potential markers in mesothelioma diagnosis include soluble mesothelin-related peptides (SMRPs) and osteopontin, but no subsequent validation has been published yet.We prospectively evaluated SMRPs in serum and pleural effusion from patients with mesothelioma (n = 74), pleural metastasis of carcinomas (n = 35), or benign pleural lesions associated with asbestos exposure (n = 28), recruited when first suspected for mesothelioma.Mean serum SMRP level was higher in patients with mesothelioma (2.05 +/- 2.57 nM/L [median +/- interquartile range]) than in patients with metastasis (1.02 +/- 1.79 nM/L) or benign lesions (0.55 +/- 0.59 nM/L). The area under the receiver operating characteristic curve (AUC) for serum SMRP was 0.872 for differentiating mesothelioma and benign lesions, cut-off = 0.93 nM/L (sensitivity = 80%, specificity = 82.6%). The AUC for serum SMRP differentiating metastasis and mesothelioma was 0.693, cut-off = 1.85 nM/L (sensitivity = 58.3%, specificity = 73.3%). SMRP values in pleural fluid were higher than in serum in all groups (mesothelioma: 46.1 +/- 83.2 nM/L; benign lesions: 6.4 +/- 11.1 nM/L; metastasis: 6.36 +/- 21.73 nM/L). The AUC for pleural SMRP-differentiating benign lesions and mesothelioma was 0.831, cut-off = 10.4 nM/L (sensitivity = 76.7%, specificity = 76.2%). The AUC for pleural SMRP-differentiating metastasis and mesothelioma was 0.793.We show that SMRPs may be a promising marker for mesothelioma diagnosis when measured either in serum or pleural fluid. The diagnostic value of SMRPs was similar in both types of samples, but pleural fluid SMRPs may better discriminate mesothelioma from pleural metastasis.
An in vitro method is described to assess the influence of synthetic calcium phosphate powders on osteoblast activities. Human osteoblast cell cultures were established from iliac crest. MC3T3-E1, an established osteogenic cell line, was employed as a control. Scanning and transmission electron microscopic observations clearly demonstrated the internalization of particles of calcium phosphate by the two osteoblast cell populations. As a consequence to the phagocytotic process, RNA transcription and protein synthesis were stimulated, as indicated by the measurements of labeled uridine, leucine and proline uptakes. From these data, it is proposed that such an in vitro model, using one of the specific cell types involved in the tissue responses to implants, could be useful to assess the biological response at the cell-biomaterial interaction.
