Europ. J. Cancer Vol. 11, pp. 91-92. Pergamon Press 1975. Printed in Great Britain
Distinctive Cytoplasmic Inclusions in Chronic Lymphocytic Leukaemia J. c. CAWLEY, J. EMMINES, A. H. GOLDSTONE, T. HAMBLIN, D. H O U G H and J. L. SMITH Department of Medicine, University of Cambridge and The Tenovus Research Laboratory, University of Southampton, Great Britain
Abstract~Distinctive cytoplasmic inclusions of a type hitherto given little attention are described in two of a series of thirty cases of chronic lymphocytic leukaemia. The possible nature and significance of these inclusions are discussed and attention is drawn to their similarity to inclusions prominent in leukaemic reticuloendotheliosis.
electron microscopy in a routine fashion [2]. Routine Leishman preparations of whole blood and buffy coat were examined in all cases.
INTRODUCTION A VARIETY of different cytoplasmic inclusions has been found in the lymphocytes of chronic lymphocytic leukaemia (CLL) [1-3]. The purpose of this paper is to report the presence in C L L of distinctive cytoplasmic inclusions of a type which has hitherto received little attention. Similar inclusions have been previously described in only one case of CLL [4] and one case of lymphosarcoma [5], although similar inclusions seem to be a prominent feature ofleukaemic reticuloendotheliosis [6-8]. These distinctive cytoplasmic inclusions were present in two of a series of 30 cases of C L L and therefore, although not as common as in hairy cell leukaemia, they are probably not particularly rare in CLL.
RESULTS The cytoplasmic inclusions were observed in two of the 30 cases studied. The inclusions could not be identified with any certainty in the Romanowsky preparations, and were only detectable during ultrastructural examination. The morphology of the inclusions is illustrated in Figs. 1 and 2. On longitudinal section (Fig. 1), they are seen to be composed of a central space flanked by two rows of approximately parallel fibrils or lamellae interspersed with ribosome-like particles. In transverse section (Fig. 2), the inclusions had a central space surrounded by several concentric lamellae. Both in longitudinal and transverse section (Figs. 1 and 2), the inclusions were frequently partially or completely surrounded by strands of rough endoplasmic reticulum. The morphology of those lymphocytes containing inclusions in no way differed from the other lymphocytes present. In both the cases of CLL, the inclusions were present in less than 10°/0 of the lymphocytes.
MATERIAL AND METHODS The cytoplasmic inclusions described in this report were observed while screening a series of 30 typical cases of C L L at the ultrastructural level for the presence of immunoglobulin crystals [9]. All cases were typical examples of C L L with regard to clinical picture, cytology and response to therapy, and none resembled leukaemic reticuloendotheliosis. Leucocytes were separated by dextran sedimentation, fixed in 3% glutaraldehyde in cacodylate-HCl buffer, and then processed for
DISCUSSION The inclusions described here closely resemble those described by Zucker Franklin [4]
Accepted 24 September 1974. 91
92
J. C. Cawley, J. Emmines, A. H. Goldstone, J. Hamblin, D. Hough and J. Smith
in a single case of C L L and those documented recently by A n d a y et al. [5] in a case of what they regarded as lymphosarcoma. The paucity of previous reports would suggest that these inclusions are rare in CLL. However, our demonstration of the inclusions in two cases of a series of 30 suggests that they may be commoner in C L L than has been recognised hitherto. Although they probably contain RNA [5], the nature and significance of the inclusions is uncertain, but it is interesting to note that similar inclusions appear to be a prominent feature in at least 50% of cases of leukaemic
reticuloendotheliosis [7]. Indeed it has been suggested that they are specific for that condition [6, 8]. However, the present study confirms the conclusion reached by Daniel and Flandrin [7] that the inclusions are present in both C L L and leukaemic reticuloendotheliosis. Since both C L L [10, 11] and leukaemic reticuloendotheliosis [12] are probably B cell diseases, it is tempting to speculate that these inclusions may represent a marker of B cells. However, why, in both C L L and leukaemic reticuloendotheliosis, only some cases, and a minority of cells in these cases, contain inclusions, remains unexplained.
REFERENCES
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11.
12.
Y. TANAKA and J. R. GOODMAN,Electron Microscopy of Human Blood Cells. Harper & Row, New York (1972). J. C. CAWLEY and F. G. J. HAYHOE, Ultrastructure of Haemic Cells. W. B. Saunders, London (1973). M. BEssls, Living Blood Cells and their Ultrastructure. Springer-Verlag, Berlin (1973). D. ZUCKER-FRANKLIN.Virus-like particles in the lymphocytes of a patient with chronic lymphocytic leukaemia. Blood 21, 509 (1963). G . J . ANDAY,J. R. GOODMANand G. H. TISHKOFF,An unusual cytoplasmic ribosomal structure in pathologic lymphocytes. Blood 41, 439 (1973). I. KATAYAMA,C. Y. LI and L. T. YAM, Ulstrastructural characteristics of the hairy "cells" of leukaemic reticuloendotheliosis. Amer. J. Path. 67, 361 (1972). M . T . DANIEL and G. FLANDRIN,Fine structure of hairy cell. Lab. Invest. 30, 1 (1974). D. CATOVSKY,J. E. PETa'IT, A. G. GALTON,A. S. D. SPIERSand C. V. HARRISON, Leukaemic reticuloendotheliosis ("hairy" cell leukaemia): a distinct clinico-pathological entity. Brit. J. Haemat. 26, 9 (1974). J . C . CAWLEY, C. R. BARKER,R. BRITCHFORDand J. L. SMITH, Crystalline inclusions in a case of chronic lymphocytic leukaemia, or. clin. exp. Immunol. 13, 407 (1972). J . L . PREUD'HOMMEand M. SELIOMANN,Surface bound immunoglobulins as a cell marker in human lympho-proliferative diseases. Blood, 40, 777 (1972). A.C. AISENBER%K. J. BLOCKand J. C. LONO, Cell-surface immunoglobulins in chronic lymphocytic leukaemia and allied disorders. Am. J. Med. 55, 184 (1973). D. CATOVSKY,J. E. PETTIT, J. GALETTO, A. OKOS and D. A. G. GALTON, The B-lymphocyte nature of the hairy cell of leukaemic reticuloendotheliosis. Brit. J. Haemat. 26, 29 (1974).
Fig. 1. (x 27,500) A typical cytoplasmic inclusion is shown in approximately longitudinal section. A central area of cytoplasm containing strands of rough endoplasmic reticulum (RER) is flanked by parallel fibrils. Numerous ribosome-like particles are scattered among these fibrils. The inclusion is partially surrounded by strands of RER ( ~ ). The general cytology of the cell is typical of the lymphocytes of CLL; part of the nucleus ( N) with heavy peripheral ehromatin condensation is shown, and the cytoplasm contains scattered mitochondria, pinocytotic vesicles, RER and free ribosomes. Fig. 2. (x 27,500) Two inclusions sectioned transversely are shown. The central area of cytoplasm is enclosed by concentric fibrils interpersed with ribosome-like particles. The inclusion is enclosed, both within and without, by strands of RER. The nucleus with heavily condensedperipheral chromatin is typical of the small lymphocyte of CLL.
(to face p. 92)
Distinctive Cytoplasmic Inclusions in Chronic Lymphocytic Leukaemia J. c. CAWLEY, J. EMMINES, A. H. GOLDSTONE, T. HAMBLIN, D. H O U G H and J. L. SMITH Department of Medicine, University of Cambridge and The Tenovus Research Laboratory, University of Southampton, Great Britain
Abstract~Distinctive cytoplasmic inclusions of a type hitherto given little attention are described in two of a series of thirty cases of chronic lymphocytic leukaemia. The possible nature and significance of these inclusions are discussed and attention is drawn to their similarity to inclusions prominent in leukaemic reticuloendotheliosis.
electron microscopy in a routine fashion [2]. Routine Leishman preparations of whole blood and buffy coat were examined in all cases.
INTRODUCTION A VARIETY of different cytoplasmic inclusions has been found in the lymphocytes of chronic lymphocytic leukaemia (CLL) [1-3]. The purpose of this paper is to report the presence in C L L of distinctive cytoplasmic inclusions of a type which has hitherto received little attention. Similar inclusions have been previously described in only one case of CLL [4] and one case of lymphosarcoma [5], although similar inclusions seem to be a prominent feature ofleukaemic reticuloendotheliosis [6-8]. These distinctive cytoplasmic inclusions were present in two of a series of 30 cases of C L L and therefore, although not as common as in hairy cell leukaemia, they are probably not particularly rare in CLL.
RESULTS The cytoplasmic inclusions were observed in two of the 30 cases studied. The inclusions could not be identified with any certainty in the Romanowsky preparations, and were only detectable during ultrastructural examination. The morphology of the inclusions is illustrated in Figs. 1 and 2. On longitudinal section (Fig. 1), they are seen to be composed of a central space flanked by two rows of approximately parallel fibrils or lamellae interspersed with ribosome-like particles. In transverse section (Fig. 2), the inclusions had a central space surrounded by several concentric lamellae. Both in longitudinal and transverse section (Figs. 1 and 2), the inclusions were frequently partially or completely surrounded by strands of rough endoplasmic reticulum. The morphology of those lymphocytes containing inclusions in no way differed from the other lymphocytes present. In both the cases of CLL, the inclusions were present in less than 10°/0 of the lymphocytes.
MATERIAL AND METHODS The cytoplasmic inclusions described in this report were observed while screening a series of 30 typical cases of C L L at the ultrastructural level for the presence of immunoglobulin crystals [9]. All cases were typical examples of C L L with regard to clinical picture, cytology and response to therapy, and none resembled leukaemic reticuloendotheliosis. Leucocytes were separated by dextran sedimentation, fixed in 3% glutaraldehyde in cacodylate-HCl buffer, and then processed for
DISCUSSION The inclusions described here closely resemble those described by Zucker Franklin [4]
Accepted 24 September 1974. 91
92
J. C. Cawley, J. Emmines, A. H. Goldstone, J. Hamblin, D. Hough and J. Smith
in a single case of C L L and those documented recently by A n d a y et al. [5] in a case of what they regarded as lymphosarcoma. The paucity of previous reports would suggest that these inclusions are rare in CLL. However, our demonstration of the inclusions in two cases of a series of 30 suggests that they may be commoner in C L L than has been recognised hitherto. Although they probably contain RNA [5], the nature and significance of the inclusions is uncertain, but it is interesting to note that similar inclusions appear to be a prominent feature in at least 50% of cases of leukaemic
reticuloendotheliosis [7]. Indeed it has been suggested that they are specific for that condition [6, 8]. However, the present study confirms the conclusion reached by Daniel and Flandrin [7] that the inclusions are present in both C L L and leukaemic reticuloendotheliosis. Since both C L L [10, 11] and leukaemic reticuloendotheliosis [12] are probably B cell diseases, it is tempting to speculate that these inclusions may represent a marker of B cells. However, why, in both C L L and leukaemic reticuloendotheliosis, only some cases, and a minority of cells in these cases, contain inclusions, remains unexplained.
REFERENCES
1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11.
12.
Y. TANAKA and J. R. GOODMAN,Electron Microscopy of Human Blood Cells. Harper & Row, New York (1972). J. C. CAWLEY and F. G. J. HAYHOE, Ultrastructure of Haemic Cells. W. B. Saunders, London (1973). M. BEssls, Living Blood Cells and their Ultrastructure. Springer-Verlag, Berlin (1973). D. ZUCKER-FRANKLIN.Virus-like particles in the lymphocytes of a patient with chronic lymphocytic leukaemia. Blood 21, 509 (1963). G . J . ANDAY,J. R. GOODMANand G. H. TISHKOFF,An unusual cytoplasmic ribosomal structure in pathologic lymphocytes. Blood 41, 439 (1973). I. KATAYAMA,C. Y. LI and L. T. YAM, Ulstrastructural characteristics of the hairy "cells" of leukaemic reticuloendotheliosis. Amer. J. Path. 67, 361 (1972). M . T . DANIEL and G. FLANDRIN,Fine structure of hairy cell. Lab. Invest. 30, 1 (1974). D. CATOVSKY,J. E. PETa'IT, A. G. GALTON,A. S. D. SPIERSand C. V. HARRISON, Leukaemic reticuloendotheliosis ("hairy" cell leukaemia): a distinct clinico-pathological entity. Brit. J. Haemat. 26, 9 (1974). J . C . CAWLEY, C. R. BARKER,R. BRITCHFORDand J. L. SMITH, Crystalline inclusions in a case of chronic lymphocytic leukaemia, or. clin. exp. Immunol. 13, 407 (1972). J . L . PREUD'HOMMEand M. SELIOMANN,Surface bound immunoglobulins as a cell marker in human lympho-proliferative diseases. Blood, 40, 777 (1972). A.C. AISENBER%K. J. BLOCKand J. C. LONO, Cell-surface immunoglobulins in chronic lymphocytic leukaemia and allied disorders. Am. J. Med. 55, 184 (1973). D. CATOVSKY,J. E. PETTIT, J. GALETTO, A. OKOS and D. A. G. GALTON, The B-lymphocyte nature of the hairy cell of leukaemic reticuloendotheliosis. Brit. J. Haemat. 26, 29 (1974).
Fig. 1. (x 27,500) A typical cytoplasmic inclusion is shown in approximately longitudinal section. A central area of cytoplasm containing strands of rough endoplasmic reticulum (RER) is flanked by parallel fibrils. Numerous ribosome-like particles are scattered among these fibrils. The inclusion is partially surrounded by strands of RER ( ~ ). The general cytology of the cell is typical of the lymphocytes of CLL; part of the nucleus ( N) with heavy peripheral ehromatin condensation is shown, and the cytoplasm contains scattered mitochondria, pinocytotic vesicles, RER and free ribosomes. Fig. 2. (x 27,500) Two inclusions sectioned transversely are shown. The central area of cytoplasm is enclosed by concentric fibrils interpersed with ribosome-like particles. The inclusion is enclosed, both within and without, by strands of RER. The nucleus with heavily condensedperipheral chromatin is typical of the small lymphocyte of CLL.
(to face p. 92)
