Fluorescing Particles (F-Bodies) in Lymphocytes in Chronic

PRELIMINARY COMMUNICATION

Scand J Haematol(l980) 25,93-95

Fluorescing Particles (F-Bodies) in Lymphocytes in Chronic Lymphocytic Leukaemia Resembling Y-Bodies IDA MARIA LISSE,J0RGEN L. THOMSEN & NIELSHENFUK HOLLANDER Department of Pathology and Division ofHaematology, Department of Medicine, Hvidovre University Hospital of Copenhagen, Hvidovre, Denmark The finding of fluorescing Y-body-like particles (F-bodies) in lymphocytes in patients with chronic lymphocytic leukaemia is reported. This finding may have diagnostic and pathogenetic implications. Key words: F-bodies- fluorescingparticles - Y-bodies- Y-chromosomes Accepted for publication July 20,1980 Correspondence to: Dr. Ida Maria Lisse, Department of Pathology, Hvidovre Hospital, DK-2650 Hvidovre, Denmark

The technique fqr Y-chromosome detection in interphase nuclei has been known for a decade as it was originally described by Pearson et a1 (1970). It has proven valuable in the field of genetics, forensic medicine etc. for sex determination and detection of Y-chromosome abnormalities such as XYY. We report on the finding of fluorescent Y-body-like particles (F-bodies) in lymphocytes in patients with chronic lymphocytic leukaemia (CLL). METHOD Blood with anticoagulant (K2EDTA) added was smeared on slides, air dried, fixed in Carnoy’s solution for a minimum of 30 min and stained in Quinacrine Dihydrochloride (Thomsen 1975). RESULTS

Our observation was an incidental finding of F-bodies in 30 % of mononuclear (MN) cells

in blood from a female with CLL (Figure 1 ~ ) No . F-bodies were observed in polymorphonuclear (PMN) leucocytes. A normal XX karyotype was found after stimulation of leucocytes from this patient with phytohaemagglutinin. The Trypsin-Giemsa and Quinacrine banding techniques were used. No abnormal fluorescing part that would explain the observation was found on any of the chromosomes. However, using this technique only T-lymphocytes are stimulated, and this does not exclude chromosome abnormalities in B-lymphocytes, the predominant cell in CLL. In a case of CLL in a female Chrz et a1 (1972) has described the finding of fluorescing bodies. We are not avare of any other reports on this. Figure 2 shows the results of F-body detection in MN cells in blood smears from 4 females (including the one described above)

0036553X/80/060093-03$02.50/0 0 1980 Munksgaard, Copenhagen

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1. M. LISSE, J. L. THOMSEN & N. H. HOLLANDER

. from female with CLL showing Figure l ~ Lymphocyte fluorescing particle indistinguishablefrom a Y-chromosome.

Figure 1 ~Lymphocyte . from male with CLL showing 2 fluorescing particles.

with CLL (mean age: 69 years), a control material of 6 females without haematologic disorders (mean age: 72 years), 5 males with CLL (mean age: 71 years), and 4 males without haematologic disorders (mean age: No of F-bodies 75 years). The detection was performed in per 50 MN blind trials. It appears that the number of F-bodies was much higher in patients with CLL. 75 a 3 out of 4 females with CLL had a F-body frequency well above the discrimination limit a of 10 % between males and females. This is a a very rare occurrence in normal females (Kringsholm et a1 1977). These F-bodies were indistinguishable from normal small Y-chromosomes. More than one fluorescing particle was seen in 34% of MN cells from males with CLL as compared to 2 % in non-CLL males (Figure 1 ~ ) Usually . there were 2, but 1 male had 4 cells with triple bodies and 1 cell with 5 fluorescent bodies. The bodies differed in size. In all of the cases a F-body count was also performed in PMN I ? I I d 1 leucocytes. The results were in accordance with the phenotypic sex of the persons with Figure 2. The number of flu0rescent.F-bodies per 50 MN cells in patients with CLL as compared to controls. the exception of 1 non-CLL male who was The male-female discrimination limit 10 % is depicted. false negative. It is, however, known that See text for further explanation. Y-body detection in PMN leucocytes is more

f

EXTRA FLUORESCING BODIES IN CLL

difficult than in MN cells (Thomsen 1979). The findings described seem to be restricted to lymphocytes, indicating an acquired anomaly. Chromosome abnormalities in haematologic diseases are frequently observed, e.g. the Philadelphia chromosome. DISCUSSION

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selectively stimulating B-lymphocytes etc. with the hope of revealing the nature of the anomaly. REFERENCES Chrz F, Michalovl K, Benesovl E & Possnerovl V (1972) A finding of F body in blood cells of a woman with karyotype 46 XX with chronic lymphatic leukemia. Lek 74,218-21. a i n g s h o h B, Thomsen J L 8~ Henningsen K (1977) ~ Fluorescent ~ ~ ~Y-chromosomes in hairs and blood stains. Forensic Sci 9, 117-26. Pearson L, Bobrow Vosa (1970) Technique

The anomaly we have observed is probably abnormal D N A - of~intra~ or~ extra~ cellular origin. This could for instance refleet a chromosome abnormality and/or a for identifying Y-chromosomes in human interphase virus particle. Our investigations so far do nuclei. Nature 226,78430. not allow a conclusion regarding this. We Thomsen J L (1975) Eine verbesserte Methode zum Nachweis des Y-chromosoms in Blutspuren. Z believe that the finding may show aspects

with regard to the pathogenesis Of cLL and it may further have diagnostic imp1icationsWork is in progress with the purpose of

Rechtsmed 76,81436. Thomsen J L (1979) Y-chromosome detection in blood stains. 8th Int Congr Soc Forensic Haemagenetics, p 183, London Sept 23-27.