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New modified method for analysis of cytrometry diploid DNA histogram by image cytrometry in chronic lymphoproliferative disorders (CLPD)

CLPD are typically low-grade neoplasms with a diploid DNA index and low proliferative activity with cells arrested in Go phase of cell cycle. Aim of the study: 1. to evaluate new modified method for analysis of diploid DNA histogram ; 2. to correlate proliferative activity including new parameters of diploid DNA histogram with lymphocyte size doubling time (DTL) ; tumor mass size doubling time (DTM), high or low total tumor mass (TTM) and survival. Patients and methods: 82 peripheral blood (PB), 87 bone marrow (BM) and 52 lymph nodes (LN) cytology smears from 155 patients were analyzed. Cytology smears stained according to Feulgen for image cytometry (ICM). Digital image analysis was performed by use of the SFORM software (VAMSTEC, Zagreb). We analyzed a total of 6 parameters for each individual lymphatic nucleus: DNA index (DI) ; percentage of cells in peak of DNA histogram (PEAK) ; percentage of cells with lower contents of DNA than cells in peak of DNA histogram(<PEAK) ; percentage of cells with higher contents DNA than cells in peak of DNA histogram (>PEAK) ; percentage of cells with higher contents DNA than 4N (>4N) and percentage of cells in S-phase (SFC). Results: Significantly better survival is found in patients with cells in Go/G1-PEAK between 28, 2-47%, >20% of cells with contents DNA<PEAK ; >4, 1 % of cells in SFC (BM lymphatic cells) ; DI> 1, 01 ; <45% of cells with contents DNA>PEAK (PB lymphatic cells) ; >35% of cells in Go/G1-PEAK and <2, 5 % of cells with contents DNA>4N (LN lymphatic cells). DTM has shown statistically significant differences (p<0, 05) for % of cells in SFC (PB lymphatic cells) ; DTL for % of cells with contents DNA>PEAK (PB lymphatic cells) ; high and low TTM for both % of cells in SFC and % of cells with contents DNA>PEAK (PB lymphatic cells) and % of cells with contents DNA>PEAK (LN lymphatic cells). Conclusion: The results obtained demonstrate that new parameters of diploid DNA histogram correlate with survival and prognostic factors (DTM and DTL) as well as assessment of TTM in patients with CLPD. The material of diploid tumor is sufficient for cell kinetic analysis by ICM with new modified analysis of diploid DNA histogram.

Chronic lymphoproliferative disorders; diploid DNA histogram

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