# FLOW CYTOMETRIC ANALYSIS OF DNA IN HUMAN LYMPHOMA CELLS

> **NIH NIH R01** · UNIVERSITY OF FLORIDA · 1987 · $130,725

## Abstract

Flow cytometry (CFM) allows the rapid measurement of antigens and nucleic
acids in individual cells.  DNA analysis by FCM is particularly useful
because it provides rapid information on cell cycle kinetics and tumor
ploidy.  B-cell lymphomas are kinetically heterogeneous tumors composed of
immunoglobulin-bearing lymphocytes.  Curiously, these tumor often contain
large numbers of normal T-cells which hamper the precise analysis of DNA in
tumor B-cells.  By labelling cell surface immunoglobulins, we measured DNA
in the neoplastic cells of B-cell lymphomas without interference from
T-cells.  The DNA synthetic fraction for the neoplastic cells correlated
remarkabley well with histologic type and it should be an useful indicator
of tumor proliferative activity.  By comparing the DNA content of tumor
B-cells with that of non-neoplastic T-cells from the same tissue, we
detected minimal (2%) aneuploidy in neoplastic cells.  We also observed two
neoplastic populations with different DNA content in approximately 10% of
the tumors suggesting ciclonality.  In the present study we propose to
answer the following:  What is the nature of aneuploidy detected by FCM in
the B-cell lymphomas?  Does it only reflect genomic size or is it in part
due to binding properties of the fluorescent dyes to neoplastic or cycling
cells?  Is the conversion of low grade lymphomas into high grade lymphomas
accompanied by ploid or kinetic changes?  What is the nature of the
"biclonality" observed in 10% of B-cell lymphomas?  Since most of the
second "clones" in these cases are near-tetraploid, do they represent
arrested or cycling cells?  If they are cycling, is the DNA synthetic rate
of the aneuploid cells higher or lower than that of the diploid cells of
the same tumor?  Are multiple "clones" related to previous therapy?  Is
there a relationship between T-cell subset distribution within the lymph
node and the kinetic properties of the neoplastic B-cells?  We plan to
answer these questions as follows:  Modal chromosome number determined on
metaphases obtained in direct preparations of tumor cells or in normal and
tumor cells induced by stimulation with B-cell mitogens will be compared
with the DNA index obtained by FCM.  Various DNA fluorescent probes will be
tested to determine if ploidy changes reflect DNA binding properties of the
fluorescent dyes.  DNA indices and kinetics properties of recurrent tumors
will be compared with those obtained in previous biopsies.  Simultaneous
analysis of cellular DNA content and bromodeoxyuridine incorporation will be
performed to establish if aneuploid cells are cycling and their rate of DNA
synthesis.  Correlations will be made between therapy prior to biopsy and
tumor ploidy.  T-lymphocyte subsets within the tumors will be analyzed to
assess if a relationship exists between T-cell distribution and kinetic
properties of the neoplastic B-cells.

## Key facts

- **NIH application ID:** 3177654
- **Project number:** 5R01CA039022-03
- **Recipient organization:** UNIVERSITY OF FLORIDA
- **SAM.gov UEI:** NNFQH1JAPEP3
- **Principal Investigator:** RAUL C. BRAYLAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $130,725
- **Award type:** 5
- **Project period:** 1984-12-01 → 1989-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3177654

## Citation

> US National Institutes of Health, RePORTER application 3177654, FLOW CYTOMETRIC ANALYSIS OF DNA IN HUMAN LYMPHOMA CELLS (5R01CA039022-03). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3177654. Licensed CC0.

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