Scand J Haematol(1979) 23, 51-54
Three Different Acid Phosphatase Patterns in Leukaemic Lymphoid T-Cells HELENA SC~NDERGMRD-PETERSEN & ANNEMARIE BOESEN 1
University Departments of Internal Medicine 11 (Haematology) and 2 Pathology, Amtssygehuset, Aarhus, Denmark
The acid phosphatase pattern was studied in leukaemic cells from 8 patients with T-cell leukaemia (5 ALL and 3 CLL). In 2 cases the enzyme activity was focal granular with paranuclear localization as earlier demonstrated by other authors, while - in contrast to these findings - the enzyme activity in 4 cases demonstrated universal granular distribution. Almost all the cells from each patient showed the same picture. I n the last 2 cases a mixed focal and universal granular pattern was observed, where half the cells possessed the focal form and the other half the universal form of granular activity. The two first-mentioned patterns were observed in cases of T-ALL as well as of T-CLL, while the mixed pattern was seen only in cases of T-ALL. Key words: acid phosphatase
- lymphocytic
leukaemia
- T-cells
Accepted fosr publication May 1, 1979 Correspondence to: Dr. Anne Marie Boesen, Institute of Pathology, Amtssygehuset, DK-8000 Aarhus C. Denmark
During the last years, there have been several reports of a distinct positive reaction of the acid phosphatase enzyme in leukaemic lymphoid cells of T-cell origin, especially with regard to the blast cells of Tlymphoblastic leukaemia (T-ALL) (Catovsky et a1 1974, 1978, Ritter et a1 1975, Brouet et a1 1976, Huhn et a1 1976ah). In these papers the characteristic strong activity of acid phosphatase is described as a dense spot-like structure with paranuclear localization, at the ultrastructural level demonstrated in membranes of the Golgi apparatus and in lysosomal granules
(Catovsky et a1 1975). Our observations suggest that another acid phosphatase pattern in leukaemic Tcells is equally typical. MAWRIAL AND METHODS Bone marrow and blood cells, obtained from 5 patients with acute lymphoblastic leukaemia and 3 patients with chronic lymphocytic leukaemia, all of T-cell origin (Table l), were studid.*
*
The examination of the lymphocyte subpopulations was performeld by Dr. J. Ellegaard, Department of Medicine and Haematology, A mt s sygehuset, Aarhus, Denmark.
0036-553W79/060051-04 $02.50/0 @ 1979 Munksgaard, Copenhagen
H. SQNDERGAARD-PETERSEN & A. M. BOESEN
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ACID PHOSPHATASE IN LEUKAEMIC T- CELLS
Figure 1. Focal, granular acid phosphatase activity - spot-like (Case 5).
Figure 2. Focal, granular acid phosphatase activity - a focal collection of granules (Case 2).
The acid phosphatase activity was demonstrated using the method described by Goldberg & Barka (1962) with the modifications worked out by Leder & Stutte (1975): Unfixed air-dried smears; naphthol-AS-BI-phosphate dissolved in dimethylformamid as a substrate and hexazonium pararosanilin (4% solution in 2 N HCI with 4 % solution of sodium nitrite) as a coupler. The slides were incubated for 4 h, rinsed in tap water, counterstained with haemalum and mounted in glycerin-gelatine. Spontaneous rosette formation with SRBC was used as a T-cell marker by employing the technique described by Jondal et a1 (1972).
53
Figure 3. Universal, granular acid phosphatase activity (Case 1).
ly to most of or to the whole cytoplasm (universal, granular distribution) (Figure 3). In the last 2 cases we observed a mixed pattern (mixed focal and universal granular distribution) in 98-100 % of the cells, in such a way that half the cells showed focal granular activity and the other half the universal granular distribution. The two first mentioned patterns were demonstrated in cases of T-ALL as well as of T-CLL, while the last pattern was seen only in cases of T-ALL.
RESULTS
The results obtained are presented in Table 1. A distinct positive reaction of acid phosphatase was present in more than 80 % of the leukaemic cells from these patients, but in different patterns. In two cases, the enzyme activity was detectable in 88-97 % of the cells as granular or spot-like deposits, confined to the paranuclear region (focal, granular distribubution) (Figures 1 and 2). In 4 cases, the activity in 89-100 % of the cells was seen as granular deposits, extending perinuclear-
COMMENTS
Until now it has been claimed that only distinct focal and paranuclear activity of acid phosphatase is diagnostic as a T-cell marker in lymphoblasts. In our experience, universal granular enzyme activity can also be taken as evidence that the leukaemic cells from patients with ALL and CLL are of T-cell origin. It might be emphasized that the enzyme activity in these cases of ALL and CLL is markedly granular, unlike the weak, mainly diffuse, activity seen
54
H. SQNDERGAARD-PETERSEN & A. M. BOESEN
in cases of CLL of the ‘common’ B-type and in cases of ‘non-T-non-B’ ALL. In normal blood lymphocytes from 120 persons aged 4 to 70 years (unpublished data), we have - concerning cytochemical demonstration of acid phosphatase activity - observed a very heterogeneous picture, including cells with no acid phosphatase activity at all, cells with diffuse, diffuse and granular, and finally granular activity alone in varying intensity and distribution quite unlike the very uniform pattern demonstrated in the above mentioned leukaemic cells. Monocytes, normal as leukaemic, possess a mixed diffuse and granular activity, are easy to distinguish morphologically and cytochemically from other cell populations, and do not make up any differential diagnostic problem. REFERENCES Brouet J-C, Valensi F, Daniel M-T, Flandrin G, Preud’Homme J L & Seligman M (1976) Immunological classification of acute lymphoblastic leukaemias. Evaluation of its clinical significance in a hundred patients. Br J Haemato1 33, 319-28. Catovsky D, Frisch B & van Noorden S (1975) B, T and ‘null’ cell leukaemias (electron cytochemistry and surface morphology). Blood
Cells 1, 115-24. Catovsky D, Galetto J, Okos A, Miliani E & Galton D A G (1974) Cytochemical profile of B and T leukaemia lymphocytes with special reference to acute lymphoblastic leukaemia. J Clin Pathol 27, 767-71. Catovsky D, Greaves M F, Pain C, Cherchi M, Janossy G & Kay H E M (1978) Acid phosphatase reaction in acute lymphoblastic leukaemia. Lancet i, 749-51. Goldberg A F & Barka T (1962) Acid phosphatase activity in human blood cells. Nature 195, 297. Huhn D, Rodt H & Thiel E (1976a) Acid phosphatase in acute lymphatic leukaemia (ALL). Immunological diagnosis of leukaemias and lymphomas, pp 169-70. Springer Verlag, Berlin J Heidelberg / New York. Huhn D, Rodt H, Thiel E, Grosse-Welde H, Fink U, Thelm H, Jager G, Steidle C & Thierfelder S (1976b) T-Zell-Leukamien des Envachsenen. Blut 33, 141-60. Jondal M, Holm G & Wigzell H (1972) Surface markers on human B and T lymphocytes. I. A large population of lymphocytes forming nonimmune rosettes with sheep red blood cells. J Exp Med 136, 207-15. Leder L-D & Stutte H J (1975) Seminar fur hamatologisch-zytochemische Techniken. Verh Dtsch Ges Pathol 59, 503-09. Ritter J, Gaedicke G, Winkler K & Landbeck G (1975) Immunologische Oberflachenmarker von Lymphoblasten bei der saure Phosphatase positiven akuten Leukaemia. Z Kinderheilkd 120, 211-15.
Three Different Acid Phosphatase Patterns in Leukaemic Lymphoid T-Cells HELENA SC~NDERGMRD-PETERSEN & ANNEMARIE BOESEN 1
University Departments of Internal Medicine 11 (Haematology) and 2 Pathology, Amtssygehuset, Aarhus, Denmark
The acid phosphatase pattern was studied in leukaemic cells from 8 patients with T-cell leukaemia (5 ALL and 3 CLL). In 2 cases the enzyme activity was focal granular with paranuclear localization as earlier demonstrated by other authors, while - in contrast to these findings - the enzyme activity in 4 cases demonstrated universal granular distribution. Almost all the cells from each patient showed the same picture. I n the last 2 cases a mixed focal and universal granular pattern was observed, where half the cells possessed the focal form and the other half the universal form of granular activity. The two first-mentioned patterns were observed in cases of T-ALL as well as of T-CLL, while the mixed pattern was seen only in cases of T-ALL. Key words: acid phosphatase
- lymphocytic
leukaemia
- T-cells
Accepted fosr publication May 1, 1979 Correspondence to: Dr. Anne Marie Boesen, Institute of Pathology, Amtssygehuset, DK-8000 Aarhus C. Denmark
During the last years, there have been several reports of a distinct positive reaction of the acid phosphatase enzyme in leukaemic lymphoid cells of T-cell origin, especially with regard to the blast cells of Tlymphoblastic leukaemia (T-ALL) (Catovsky et a1 1974, 1978, Ritter et a1 1975, Brouet et a1 1976, Huhn et a1 1976ah). In these papers the characteristic strong activity of acid phosphatase is described as a dense spot-like structure with paranuclear localization, at the ultrastructural level demonstrated in membranes of the Golgi apparatus and in lysosomal granules
(Catovsky et a1 1975). Our observations suggest that another acid phosphatase pattern in leukaemic Tcells is equally typical. MAWRIAL AND METHODS Bone marrow and blood cells, obtained from 5 patients with acute lymphoblastic leukaemia and 3 patients with chronic lymphocytic leukaemia, all of T-cell origin (Table l), were studid.*
*
The examination of the lymphocyte subpopulations was performeld by Dr. J. Ellegaard, Department of Medicine and Haematology, A mt s sygehuset, Aarhus, Denmark.
0036-553W79/060051-04 $02.50/0 @ 1979 Munksgaard, Copenhagen
H. SQNDERGAARD-PETERSEN & A. M. BOESEN
52
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c
m
?
% 4
8
W
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m
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3
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F4
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cl cl
u
=3u
W
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.
$2
W
ACID PHOSPHATASE IN LEUKAEMIC T- CELLS
Figure 1. Focal, granular acid phosphatase activity - spot-like (Case 5).
Figure 2. Focal, granular acid phosphatase activity - a focal collection of granules (Case 2).
The acid phosphatase activity was demonstrated using the method described by Goldberg & Barka (1962) with the modifications worked out by Leder & Stutte (1975): Unfixed air-dried smears; naphthol-AS-BI-phosphate dissolved in dimethylformamid as a substrate and hexazonium pararosanilin (4% solution in 2 N HCI with 4 % solution of sodium nitrite) as a coupler. The slides were incubated for 4 h, rinsed in tap water, counterstained with haemalum and mounted in glycerin-gelatine. Spontaneous rosette formation with SRBC was used as a T-cell marker by employing the technique described by Jondal et a1 (1972).
53
Figure 3. Universal, granular acid phosphatase activity (Case 1).
ly to most of or to the whole cytoplasm (universal, granular distribution) (Figure 3). In the last 2 cases we observed a mixed pattern (mixed focal and universal granular distribution) in 98-100 % of the cells, in such a way that half the cells showed focal granular activity and the other half the universal granular distribution. The two first mentioned patterns were demonstrated in cases of T-ALL as well as of T-CLL, while the last pattern was seen only in cases of T-ALL.
RESULTS
The results obtained are presented in Table 1. A distinct positive reaction of acid phosphatase was present in more than 80 % of the leukaemic cells from these patients, but in different patterns. In two cases, the enzyme activity was detectable in 88-97 % of the cells as granular or spot-like deposits, confined to the paranuclear region (focal, granular distribubution) (Figures 1 and 2). In 4 cases, the activity in 89-100 % of the cells was seen as granular deposits, extending perinuclear-
COMMENTS
Until now it has been claimed that only distinct focal and paranuclear activity of acid phosphatase is diagnostic as a T-cell marker in lymphoblasts. In our experience, universal granular enzyme activity can also be taken as evidence that the leukaemic cells from patients with ALL and CLL are of T-cell origin. It might be emphasized that the enzyme activity in these cases of ALL and CLL is markedly granular, unlike the weak, mainly diffuse, activity seen
54
H. SQNDERGAARD-PETERSEN & A. M. BOESEN
in cases of CLL of the ‘common’ B-type and in cases of ‘non-T-non-B’ ALL. In normal blood lymphocytes from 120 persons aged 4 to 70 years (unpublished data), we have - concerning cytochemical demonstration of acid phosphatase activity - observed a very heterogeneous picture, including cells with no acid phosphatase activity at all, cells with diffuse, diffuse and granular, and finally granular activity alone in varying intensity and distribution quite unlike the very uniform pattern demonstrated in the above mentioned leukaemic cells. Monocytes, normal as leukaemic, possess a mixed diffuse and granular activity, are easy to distinguish morphologically and cytochemically from other cell populations, and do not make up any differential diagnostic problem. REFERENCES Brouet J-C, Valensi F, Daniel M-T, Flandrin G, Preud’Homme J L & Seligman M (1976) Immunological classification of acute lymphoblastic leukaemias. Evaluation of its clinical significance in a hundred patients. Br J Haemato1 33, 319-28. Catovsky D, Frisch B & van Noorden S (1975) B, T and ‘null’ cell leukaemias (electron cytochemistry and surface morphology). Blood
Cells 1, 115-24. Catovsky D, Galetto J, Okos A, Miliani E & Galton D A G (1974) Cytochemical profile of B and T leukaemia lymphocytes with special reference to acute lymphoblastic leukaemia. J Clin Pathol 27, 767-71. Catovsky D, Greaves M F, Pain C, Cherchi M, Janossy G & Kay H E M (1978) Acid phosphatase reaction in acute lymphoblastic leukaemia. Lancet i, 749-51. Goldberg A F & Barka T (1962) Acid phosphatase activity in human blood cells. Nature 195, 297. Huhn D, Rodt H & Thiel E (1976a) Acid phosphatase in acute lymphatic leukaemia (ALL). Immunological diagnosis of leukaemias and lymphomas, pp 169-70. Springer Verlag, Berlin J Heidelberg / New York. Huhn D, Rodt H, Thiel E, Grosse-Welde H, Fink U, Thelm H, Jager G, Steidle C & Thierfelder S (1976b) T-Zell-Leukamien des Envachsenen. Blut 33, 141-60. Jondal M, Holm G & Wigzell H (1972) Surface markers on human B and T lymphocytes. I. A large population of lymphocytes forming nonimmune rosettes with sheep red blood cells. J Exp Med 136, 207-15. Leder L-D & Stutte H J (1975) Seminar fur hamatologisch-zytochemische Techniken. Verh Dtsch Ges Pathol 59, 503-09. Ritter J, Gaedicke G, Winkler K & Landbeck G (1975) Immunologische Oberflachenmarker von Lymphoblasten bei der saure Phosphatase positiven akuten Leukaemia. Z Kinderheilkd 120, 211-15.
