Table 1.
Demographic and clinical characteristics of patients with hypereosinophilia.
Figure 1.
Representative cytocentrifuge smears of two high purity eosinophil preparations obtained from peripheral blood samples.
May-Grünwald-Giemsa staining, original magnification: X 400 in the upper panels and X 1000 (immersion) in the lower panels.
Figure 2.
Binding of human recombinant tissue factor (TF) by three commercial anti-TF antibodies (2K1, 4G4 and GMA) evaluated by western blot (upper panel) and enzyme immunoassay (lower panel) methods.
Only 2K1 efficiently recognizes TF with both methods. The last lane of western blotting refers to the size markers (SM). In enzyme immunoassay experiments, data represent the mean of three different measurements.
Figure 3.
Panel A shows the western blot analysis of tissue factor (TF) in homogenate samples of purified eosinophils from 9 patients with hyperosinophilic conditions (HE) (top), purified eosinophils from 9 normal controls (N) (middle), and purified monocytes from 2 normal controls (M), purified endothelial cells from 2 samples of cell line ECV304 and purified fibroblasts from 4 samples of cell line IMR90 (bottom).
A major band with Mr of 47,000 corresponding to the native TF was found in the eosinophil homogenates from the 9 patients and the 9 controls, with a higher intensity in the former than in the latter. The intensity of the TF band was weaker in monocytes (M1, M2) than in endothelial cells (ECV304) and in fibroblasts (IMR90). Panel B shows the western blot analysis of the ubiquitary protein beta-actin, which was well represented in all patients, normal subjects and positive controls.
Figure 4.
Panel A shows real-time polymerase chain reaction (RT-PCR) analysis of tissue factor (TF) in purified eosinophils from 9 patients with hyperesinophilia (HE, red lines) and 9 normal controls (N, blue lines).
Panel B shows RT-PCR analysis of beta-actin in purified eosinophils from 9 patients with hyperesinophilia (HE, red lines) and 9 normal controls (N, blue lines). Panel C shows RT-PCR analysis of TF in purified monocytes from 4 normal controls (green lines), in 4 samples of endothelial cell line ECV304 (blue lines) and in 8 samples of fibroblast cell line IMR90 (red lines). Panel D shows RT-PCR analysis of beta-actin in purified monocytes from 4 normal controls (green lines), in 4 samples of endothelial cell line ECV304 (blue lines) and in 8 samples of fibroblast cell line IMR90 (red lines).
Table 2.
Expression of target (tissue factor) and housekeeping (beta-actin) genes in purified eosinophils obtained from 9 patients with hypereosinophilia and 9 normal controls.