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Morphometry, DNA image cytometry and analysis of nucleolar organizer regions in chronic Iymphoproliferative disorders

Background: Chronic lymphoproliferative disorders (CLPD) are defined as a heterogeneous group of malignant monoclonal lymphocyte disorders, sharing their indolence and involvement of primary lymphoid tissue in lymph nodes (LN), bone marrow (BM), spleen and peripheral blood (PB). There are many studies about digital image analysis: morphometry, assessment of argyrophilic nucleolar organizer regions (AgNOR) and image DNA cytometry (ICM) in different benign, suspicious and malignant lesions of various tissue and thus also of leukemic and lymphoma cells. Objectives: To correlate morphometry, DNA ploidy, proliferative activity and the panern of AgNOR in CLPD between the different compartments of tumor mass (lymph nodes, bone marrow and peripheral blood). Methods: Peripheral blood, bone marrow and lymph nodes archive cytology smears from IS 5 patients were analyzed. Conventional cytology-stained smears (May-Grunwald-Giemsa) were used for morphometric analysis, secondarily stained according 10 Feulgen for ICM or silver staining for AgNOR, for each. Digital image analysis was performed by use of the SFORM software (VAMSTEC Zagreb). We analyzed a total of 50 parameters for each individual lymphocytic cell and nucleus: 22 of morphometry, 6 of ICM and 26 of AgNOR. Results: Morphometric analysis of cells and nuclei has shown statistically significant differences (p < 0.05) for area and convex area between the groups studied: BM/PB, PB/LN and BM/LN. The most [CM differences (S-phase, % of cells of GO/G1 peak, % of cells < GO/G1 peak and % of cells > GO/G1 peak) were statistically significant between BM/PB, PB/LN and BM/LN (P< 0.05). Size of inhomogenic and ringed AgNOR was significantly different in relationship to nuclei. Conclusion: The results obtained demonstrate that lymphocytic cells are produced in bone marrow (lower % cells in S-phase and the highest % cells of GO/G I peak). They migrate to lymph nodes, where transformation occurs (higher % cells in S-phase and lower % in GO/GI peak). High S-phase does not necessarily mean a bener prognosis, but bener survival is seen in patients with higher % of cells in GO/G I peak in lymph nodes. Cells migration from BM to LN and between lymph nodes takes place in PB (cells with high and low proliferative activity: high % of cells in S-phase and in GO/G I peak. as well as present large cells with large nuclei). Prognosis depends on the tumor mass which is a consequence of cells proliferation. Precise typing of the disease is a prerequisite for rational therapy care.

Chronic lymphoproliferative disordersdigital image analysis; morphometry; AgNOR; image DNA cytometry

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