Argyrophilic proteins of nucleolar organizer regions (AgNORs) in fallopian tube carcinomas.

Forty-four patients diagnosed with primary cancer of the fallopian tube (PFTC) were analyzed with regard to AgNORs expression, morphological classification, clinical stage and survival rate. Twenty-seven patients were FIGO stage I, 7 FIGO II and 10 FIGO III. Histological classification of PFTC revealed 18 endometrioid types, 9 serous, 7 undifferentiated, 6 urothelial, 2 clear cell and 2 another type of (intestinal, squamous cell) cancer. Histological grading revealed 11 G1, 16 G2 and 17 G3 tumors. The number of AgNORs per nucleus ranged from 1 to 7, mean 2.54+/-0.77. The smallest number of silver stained NORs was observed in the endometrioid type (mean 2.32+/-0.62) and the biggest number of AgNORs in undifferentiated carcinoma (mean 3.05+/-0.82). There was no correlation between number of AgNORs and AgNOR area/nuclear area ratio and survival (p=0.71), histological stage or histological type of PFTC. There was a correlation between the number of AgNORs among tumors with histological grade 1 and grade 3 (p=0.023), and grade 2 and grade 3 (p=0.045). However, there was no correlation between AgNOR number and survival rate in these groups.

Quantitative analysis of three-dimensional distribution of AgNOR proteins during interphase in leukemic cells.

Acidic proteins of the nucleolar organizer regions, selectively stained by silver (AgNOR-proteins), were investigated during interphase in leukemia cells with a confocal scanning laser microscope (CSLM). Simultaneous confocal fluorescence (for specific labeling of DNA, using propidium iodide) and transmitted light microscopy combined with digital deconvolution (for the location of the AgNOR proteins in nonconfocal mode) were used. The distribution of the AgNOR proteins measured by 3D microscopy was described by their number, the volume occupation of the nucleus by the AgNOR aggregates, the distance between each AgNOR, the distance of each AgNOR to the nucleolar border, and their anisotropy. The results of the 3D analysis were compared to those obtained by conventional 2D analysis, cytogenetical analysis of metaphase nucleolar organiser regions (NORs), and cell duplication rate. The descriptive power of these 3D parameters were assessed for nine leukemic cell lines. The measurements of the 3D spatial distribution of AgNORs was a better discriminant parameter than the morphological parameters (i.e., number and volume). The 3D expression of AgNORs is also a reliable parameter for assessing proliferative activity of leukemic cells and seems to be in relation with the differentiation stage of these leukemic cells.

Interest of argyrophilic proteins nucleolar organizer regions (AgNOR) to estimate the reactivity of T cell clones against autologous malignant B-NHL cells.

The silver-stained acidic proteins of interphase nucleolar organizer regions (AgNOR) were studied to assess the reactivity of 11 T cell clones (Tcc) against autologous B-NHL cells. Tcc, derived from tumour-infiltrating T lymphocytes of seven patients, were cultured in the presence of irradiated autologous B-NHL cells with recombinant IL-2. Then the percentage of activated T cells expressing the CD25 antigen and their proliferating rate (measured by [3H]thymidine incorporation) were estimated. Simultaneously, at the end of this culture period, B-NHL cells were eliminated by using a cell-sorter, and the resulting purified T cells were studied for AgNOR expression. Tcc cultured without B-NHL cells served as controls. In eight out of the 11 Tcc, the increase in [3H]thymidine incorporation and CD25 expression paralleled the increase of AgNOR area and number. By contrast, in the three remaining Tcc, we observed a decrease (one Tcc) or an increase (two Tcc) of AgNOR parameters, whereas CD25 expression and/or [3H]thymidine incorporation remained unchanged in comparison to control cultures. We concluded that quantification of AgNOR should be a more sensitive technique than thymidine incorporation and CD25 expression for detecting the activation in vitro of T cells induced by autologous B-NHL cells.