Sezary cell leukaemia (SCL) is a mature T-cell leukaemia with characteristic cerebriform nuclei, ... more Sezary cell leukaemia (SCL) is a mature T-cell leukaemia with characteristic cerebriform nuclei, whereas Sezary syndrome (SS) involves a mature T-cell lymphoma with a similar nuclear morphology. We have examined these diseases by cytogenetics, chromosome painting and fluorescence in situ hybridization (FISH). Both diseases had complex cytogenetic abnormalities. All three cases of SCL investigated had inv(14)(q11;q32) and two had iso(8q). No case of SS had these abnormalities but, instead, iso(17q) or 17p+ was present in the three cases of SS investigated and FISH indicated loss of heterozygosity due to deletion of a region at 17p13 that included the tumour suppressor gene P53, implicating it in this malignancy. One case of SCL had iso(17q). The abnormalities of chromosomes 8 and 14 in SCL are commonly observed in T-prolymphocytic leukaemia (T-PLL) and suggest that SCL may be a variant of T-PLL rather than of SS.
SUMMARY. Seven patients presenting as an acute leukaemia caused difficulty in diagnosis. The lymp... more SUMMARY. Seven patients presenting as an acute leukaemia caused difficulty in diagnosis. The lymphoid appearance of the blast cells either initially or during treatment suggested acute lymphoid leukaemia (ALL). In each case the Philadelphia chromosome was shown to be present thus suggesting that these cases were examples of chronic myeloid leukaemia (CML) presenting in blast crisis without a detectable chronic phase.The implications of these findings are discussed and the difficulty in achieving a precise diagnosis in the acute leukaemias is emphasized. Cytogenetic analysis should be carried out whenever the type of acute leukaemia present is of critical importance.
SUMMARY. Fourteen cases of Philadelphia chromosome (Ph1) positive chronic myeloid leukaemia in bl... more SUMMARY. Fourteen cases of Philadelphia chromosome (Ph1) positive chronic myeloid leukaemia in blast transformation have been investigated using cell surface markers. Morphologically eight cases were lymphoid and the remainder myeloid in appearance. All cases were negative with surface markers for thymocytes and T and B lymphocytes. Five of the lymphoid cases reacted with an antiserum specific for acute lymphoid leukaemia (ALL) of non-T non-B type and were also weakly reactive with a lymphocyte reactive antiserum. A sixth patient, whose blast cells were anti-ALL negative (ALL-) at presentation, subsequently developed central nervous system leukaemia with anti-ALL positive (ALL+) blast cells in the CSF. In all cases the leukaemic blast cells showed greatly diminished expression of cholera toxin receptors when compared to granulocytic cells from the chronic phase of CML. This parallels weak or negligible expression of the cholera toxin receptor in ALL and AML.These results suggest that the blastic phase of CML may involve different cellular derivatives of a pluripotential stem cell in which the primary malignant/genetic changes reside. The blast crisis of CML can therefore be heterogeneous with respect to cellular expression and in a significant proportion of patients involves a cell which is by membrane markers and morphological criteria indistinguishable from that seen in the common form of ALL. In these cases the Philadelphia chromosome may be the only distinguishing cellular characteristic.
1,25-dihydroxyvitamin D3 induces monocyte-macrophage differentiation and inhibits proliferation o... more 1,25-dihydroxyvitamin D3 induces monocyte-macrophage differentiation and inhibits proliferation of cells from the human promyelocytic leukaemia cell line HL60. Similarly human bone marrow progenitor cells differentiate preferentially along the monocyte-macrophage pathway when incubated in the presence of 1,25-dihydroxyvitamin D3. We suggest that the inhibition of growth which occurs after addition of the vitamin to HL60 might be paralleled in vivo by inhibition of proliferation of leukaemic cells; also we speculate that the vitamin may be involved in the control of both monocyte-macrophage and osteoclast production in vivo.
Three recently-derived cell lines of prolymphocytic leukemia origin were studied. The phenotype o... more Three recently-derived cell lines of prolymphocytic leukemia origin were studied. The phenotype of the cells, analysed in terms of expression of immunoglobulin and other B cell maturation markers, indicates that the cells are mature B lymphocytes close to the plasma cell stage of differentiation. All three cell lines secrete IgM, and two of the three lines respond to B cell differentiation factor with an increased secretion of IgM.
The interactions between haemopoietic and stromal elements are crucial for stem cell proliferatio... more The interactions between haemopoietic and stromal elements are crucial for stem cell proliferation and differentiation. The bulk of this evidence is derived from experiments in rodents and in-vitro culture studies. We have studied the spatial relationships between the stromal and haemopoietic components and their cellular composition in histological sections of bone marrow (BM) from seven healthy fetuses, 10 normal adults and over 60 patients with acute myeloid leukaemia (AML) and chronic granulocytic leukaemia (CGL) at different stages of the disease. During the early developmental stage (16-18 weeks) fetal BM showed focal haemopoiesis with a characteristic spatial localization of haemopoiesis near bony trabeculae and around small blood vessels. In AML following the treatment-induced hypoplasia, large uniform unilocular fat cells arranged in groups designated 'structured fat' developed from scattered multilocular precursor fat cells. Early foci of haemopoietic regeneration were present almost exclusively in areas of structured fat. In the marrow of patients with CGL in blast transformation (BT) treated by intensive therapy and autografting with cryopreserved haemopoietic stem cells, the haemopoiesis was focal. Clusters of regenerating erythroid precursors or of megakaryocytes were seen in intimate contact with marrow sinusoids and granulopoietic precursors in intimate association with small blood vessels and also in close contact with the endosteal surface of bony trabeculae. We conclude that the endosteal cells, fat cells and the vascular endothelial cells comprise the critical non-haemopoietic stromal elements of human BM. The close associations observed between the regenerating haemopoietic cells and the stromal cells provide strong evidence in support of the existence of a permissive haemopoietic micro-environment in man and emphasize the structural and functional interrelationships that exists between bone, fat, the microvascular system and haemopoiesis in human bone marrow.
Due to scheduled maintenance access to the Wiley Online Library may be disrupted as follows: Satu... more Due to scheduled maintenance access to the Wiley Online Library may be disrupted as follows: Saturday, 2 October - New York 0500 EDT to 0700 EDT; London 1000 BST to 1200 BST; Singapore 1700 SGT to 1900 SGT. ... *Correspondence: Dr D. Catovsky, MRC ...
... Using only the technique for B lymphocytes we have demonstrated the formation of EAC rosettes... more ... Using only the technique for B lymphocytes we have demonstrated the formation of EAC rosettes in the majority of 'hairy' cells in case SH ; similar findings were reported in CLL lymphocytes (Pincus et al, 1972; Ross et a!, 1973). ...
Eighteen patients with chronic granulocytic leukaemia (CGL) were treated by chemoradiotherapy and... more Eighteen patients with chronic granulocytic leukaemia (CGL) were treated by chemoradiotherapy and transplantation of bone marrow (BMT) collected from their HLA-identical sibs; engraftment with donor marrow occurred in all cases. Ten of the patients had been subjected to splenectomy before BMT; recovery after BMT of granulocyte, lymphocyte and platelet numbers in the peripheral blood was more rapid in these patients than in the eight patients who retained their spleens. Acute graft-versus-host disease occurred in 12 of the 15 evaluable patients and appeared to be more severe in those who lacked their spleens at the time of transplant. Of the 12 patients surviving at follow-up times ranging from 59 to 207 weeks, six had been subjected to splenectomy before BMT and six retained their spleens. We conclude that engraftment was more rapid in the splenectomized patients and splenectomy might have increased the chance of eradicating the leukaemia, but these considerations must be balanced against the short-term and long-term risks associated with splenectomy.
ABSTRACT CASE A 67 year old English male presented with a six month history of shortness of breat... more ABSTRACT CASE A 67 year old English male presented with a six month history of shortness of breath, malaise, lethargy, fever, and night sweats. He had not suffered pruritus or alcohol-induced pain. There was no history of recent travel. He had no hepatomegaly, lymphadenopathy, splenomegaly or abnormal findings in the chest. Results of laboratory tests were: Hb 10 g/dl, MCV 89 fl, WBC 3.8 × 109/1, neutrophil count 2.5 × 109/1, lymphocyte count 1.1 × 109/1, monocyte count 0.04 × 109/1, platelet count 269 × 109/1, ESR 116 mm/hr, aspartate transferase 44IU/1 (NR<41), alanine transferase 39 IU/1 (NR < 45), alkaline phosphatase 458 IU/1 (NR 30-115), γGT 637 IU/1 (NR < 65), bilirubin 29 μm/1 (NR 3-20), albumin 28 g/1 (NR 34-50), serum iron 4μm (NR 10-30), TIBC 43.8 μm (NR 45-70), iron saturation 9%. Normal results included urea, creatinine, serum B12, red cell and serum folate, immunoglobulin concentrations and serum electrophoretic pattern. No Bence Jones protein was detected. Serology for hepatitis A and B was negative. Culture of blood, urine, and sputum yielded no pathogens. A Ziehl-Nielsen (ZN)/auramine stain of sputum was negative as was a Mantoux test. A CT scan of the abdomen showed the liver and spleen to be normal in texture; there was no lymphadenopathy. Chest radiography and a skeletal survey were normal. The peripheral blood film showed rouleaux and occasional abnormal cells. Bone marrow aspirate was difficult and yielded a poor aspirate containing cells such as are shown in Figure 1. A trephine biopsy was markedly abnormal (Figure 2). A further diagnostic procedure was performed.
Leukaemic cells from 2 patients with B-prolymphocytic leukaemia were immortalized in vitro by mea... more Leukaemic cells from 2 patients with B-prolymphocytic leukaemia were immortalized in vitro by means of Epstein-Barr virus and phorbol-ester TPA. The resulting cell lines, named JVM-2 and JVM-3, have been growing continuously in liquid culture for more than one year. JVM-2 is characterized cytogenetically by t(11;14)(q13; q32), and JVM-3 by trisomy 12. The immunoglobulin (Ig) heavy- and light-chain genes showed the same pattern of rearrangement in both lines as in the original prolymphocytes from each case. The cells from these lines showed a spectrum of morphological and immunological features corresponding to different stages of B-cell maturation. The expression of Ig, IgM-lambda in JVM-2 and IgMD-K in JVM-3, changed from a predominantly membrane pattern in the original cells to a cytoplasmic one in the cell lines. By comparison with their original progenitors, the cells from both lines showed reduced reactivity with the monoclonal antibody (MAb) FMC7, and increased expression of the antigens recognized by the MAbs OKT10, alpha-Tac, FMC53 and Ki-I. The availability of cell lines from this rare type of lymphoid leukaemia offers a potential tool for the study of molecular events associated with the expression of Ig and other antigens by neoplastic cells.
Sixteen patients with chronic granulocytic leukaemia (CGL) in blast cell transformation (BT) were... more Sixteen patients with chronic granulocytic leukaemia (CGL) in blast cell transformation (BT) were studied by means of serial bone marrow trephine biopsies (BMB). The BMB were performed during the following stages of the disease: 1 at diagnosis of BT, 2 following ablative therapy and 3 during haematological recovery following autografting. In 10 of the 16 patients, BT was recognized by the presence of a distinctive infiltration with blast cells with a single large hyperchromatic nucleolus. In four of the 16 patients, the diagnosis of BT could only be made by BMB since simultaneous marrow aspiration yielded either a dry tap or no marrow fragments; in two of these four patients the BT was focal in the BMB specimen. Following ablative therapy and during haematological regeneration after autografting, BM aspirates were unsatisfactory in most cases and they were inadequate to assess cellularity or the presence of residual blasts. In contrast, sections of BMB showed clearly whether a decrease in cellularity took place or, in some, residual blasts were still present. BMB samples obtained 2 weeks after autografting showed haemopoietic regeneration consisting of discrete foci of either erythroid, granulocytic or megakaryocytic precursor cells. We conclude that BMB is essential for diagnosing CGL in BT, for monitoring the progress of these patients after therapy and in assessing the haemopoietic regeneration following autografting.
Sezary cell leukaemia (SCL) is a mature T-cell leukaemia with characteristic cerebriform nuclei, ... more Sezary cell leukaemia (SCL) is a mature T-cell leukaemia with characteristic cerebriform nuclei, whereas Sezary syndrome (SS) involves a mature T-cell lymphoma with a similar nuclear morphology. We have examined these diseases by cytogenetics, chromosome painting and fluorescence in situ hybridization (FISH). Both diseases had complex cytogenetic abnormalities. All three cases of SCL investigated had inv(14)(q11;q32) and two had iso(8q). No case of SS had these abnormalities but, instead, iso(17q) or 17p+ was present in the three cases of SS investigated and FISH indicated loss of heterozygosity due to deletion of a region at 17p13 that included the tumour suppressor gene P53, implicating it in this malignancy. One case of SCL had iso(17q). The abnormalities of chromosomes 8 and 14 in SCL are commonly observed in T-prolymphocytic leukaemia (T-PLL) and suggest that SCL may be a variant of T-PLL rather than of SS.
SUMMARY. Seven patients presenting as an acute leukaemia caused difficulty in diagnosis. The lymp... more SUMMARY. Seven patients presenting as an acute leukaemia caused difficulty in diagnosis. The lymphoid appearance of the blast cells either initially or during treatment suggested acute lymphoid leukaemia (ALL). In each case the Philadelphia chromosome was shown to be present thus suggesting that these cases were examples of chronic myeloid leukaemia (CML) presenting in blast crisis without a detectable chronic phase.The implications of these findings are discussed and the difficulty in achieving a precise diagnosis in the acute leukaemias is emphasized. Cytogenetic analysis should be carried out whenever the type of acute leukaemia present is of critical importance.
SUMMARY. Fourteen cases of Philadelphia chromosome (Ph1) positive chronic myeloid leukaemia in bl... more SUMMARY. Fourteen cases of Philadelphia chromosome (Ph1) positive chronic myeloid leukaemia in blast transformation have been investigated using cell surface markers. Morphologically eight cases were lymphoid and the remainder myeloid in appearance. All cases were negative with surface markers for thymocytes and T and B lymphocytes. Five of the lymphoid cases reacted with an antiserum specific for acute lymphoid leukaemia (ALL) of non-T non-B type and were also weakly reactive with a lymphocyte reactive antiserum. A sixth patient, whose blast cells were anti-ALL negative (ALL-) at presentation, subsequently developed central nervous system leukaemia with anti-ALL positive (ALL+) blast cells in the CSF. In all cases the leukaemic blast cells showed greatly diminished expression of cholera toxin receptors when compared to granulocytic cells from the chronic phase of CML. This parallels weak or negligible expression of the cholera toxin receptor in ALL and AML.These results suggest that the blastic phase of CML may involve different cellular derivatives of a pluripotential stem cell in which the primary malignant/genetic changes reside. The blast crisis of CML can therefore be heterogeneous with respect to cellular expression and in a significant proportion of patients involves a cell which is by membrane markers and morphological criteria indistinguishable from that seen in the common form of ALL. In these cases the Philadelphia chromosome may be the only distinguishing cellular characteristic.
1,25-dihydroxyvitamin D3 induces monocyte-macrophage differentiation and inhibits proliferation o... more 1,25-dihydroxyvitamin D3 induces monocyte-macrophage differentiation and inhibits proliferation of cells from the human promyelocytic leukaemia cell line HL60. Similarly human bone marrow progenitor cells differentiate preferentially along the monocyte-macrophage pathway when incubated in the presence of 1,25-dihydroxyvitamin D3. We suggest that the inhibition of growth which occurs after addition of the vitamin to HL60 might be paralleled in vivo by inhibition of proliferation of leukaemic cells; also we speculate that the vitamin may be involved in the control of both monocyte-macrophage and osteoclast production in vivo.
Three recently-derived cell lines of prolymphocytic leukemia origin were studied. The phenotype o... more Three recently-derived cell lines of prolymphocytic leukemia origin were studied. The phenotype of the cells, analysed in terms of expression of immunoglobulin and other B cell maturation markers, indicates that the cells are mature B lymphocytes close to the plasma cell stage of differentiation. All three cell lines secrete IgM, and two of the three lines respond to B cell differentiation factor with an increased secretion of IgM.
The interactions between haemopoietic and stromal elements are crucial for stem cell proliferatio... more The interactions between haemopoietic and stromal elements are crucial for stem cell proliferation and differentiation. The bulk of this evidence is derived from experiments in rodents and in-vitro culture studies. We have studied the spatial relationships between the stromal and haemopoietic components and their cellular composition in histological sections of bone marrow (BM) from seven healthy fetuses, 10 normal adults and over 60 patients with acute myeloid leukaemia (AML) and chronic granulocytic leukaemia (CGL) at different stages of the disease. During the early developmental stage (16-18 weeks) fetal BM showed focal haemopoiesis with a characteristic spatial localization of haemopoiesis near bony trabeculae and around small blood vessels. In AML following the treatment-induced hypoplasia, large uniform unilocular fat cells arranged in groups designated 'structured fat' developed from scattered multilocular precursor fat cells. Early foci of haemopoietic regeneration were present almost exclusively in areas of structured fat. In the marrow of patients with CGL in blast transformation (BT) treated by intensive therapy and autografting with cryopreserved haemopoietic stem cells, the haemopoiesis was focal. Clusters of regenerating erythroid precursors or of megakaryocytes were seen in intimate contact with marrow sinusoids and granulopoietic precursors in intimate association with small blood vessels and also in close contact with the endosteal surface of bony trabeculae. We conclude that the endosteal cells, fat cells and the vascular endothelial cells comprise the critical non-haemopoietic stromal elements of human BM. The close associations observed between the regenerating haemopoietic cells and the stromal cells provide strong evidence in support of the existence of a permissive haemopoietic micro-environment in man and emphasize the structural and functional interrelationships that exists between bone, fat, the microvascular system and haemopoiesis in human bone marrow.
Due to scheduled maintenance access to the Wiley Online Library may be disrupted as follows: Satu... more Due to scheduled maintenance access to the Wiley Online Library may be disrupted as follows: Saturday, 2 October - New York 0500 EDT to 0700 EDT; London 1000 BST to 1200 BST; Singapore 1700 SGT to 1900 SGT. ... *Correspondence: Dr D. Catovsky, MRC ...
... Using only the technique for B lymphocytes we have demonstrated the formation of EAC rosettes... more ... Using only the technique for B lymphocytes we have demonstrated the formation of EAC rosettes in the majority of 'hairy' cells in case SH ; similar findings were reported in CLL lymphocytes (Pincus et al, 1972; Ross et a!, 1973). ...
Eighteen patients with chronic granulocytic leukaemia (CGL) were treated by chemoradiotherapy and... more Eighteen patients with chronic granulocytic leukaemia (CGL) were treated by chemoradiotherapy and transplantation of bone marrow (BMT) collected from their HLA-identical sibs; engraftment with donor marrow occurred in all cases. Ten of the patients had been subjected to splenectomy before BMT; recovery after BMT of granulocyte, lymphocyte and platelet numbers in the peripheral blood was more rapid in these patients than in the eight patients who retained their spleens. Acute graft-versus-host disease occurred in 12 of the 15 evaluable patients and appeared to be more severe in those who lacked their spleens at the time of transplant. Of the 12 patients surviving at follow-up times ranging from 59 to 207 weeks, six had been subjected to splenectomy before BMT and six retained their spleens. We conclude that engraftment was more rapid in the splenectomized patients and splenectomy might have increased the chance of eradicating the leukaemia, but these considerations must be balanced against the short-term and long-term risks associated with splenectomy.
ABSTRACT CASE A 67 year old English male presented with a six month history of shortness of breat... more ABSTRACT CASE A 67 year old English male presented with a six month history of shortness of breath, malaise, lethargy, fever, and night sweats. He had not suffered pruritus or alcohol-induced pain. There was no history of recent travel. He had no hepatomegaly, lymphadenopathy, splenomegaly or abnormal findings in the chest. Results of laboratory tests were: Hb 10 g/dl, MCV 89 fl, WBC 3.8 × 109/1, neutrophil count 2.5 × 109/1, lymphocyte count 1.1 × 109/1, monocyte count 0.04 × 109/1, platelet count 269 × 109/1, ESR 116 mm/hr, aspartate transferase 44IU/1 (NR<41), alanine transferase 39 IU/1 (NR < 45), alkaline phosphatase 458 IU/1 (NR 30-115), γGT 637 IU/1 (NR < 65), bilirubin 29 μm/1 (NR 3-20), albumin 28 g/1 (NR 34-50), serum iron 4μm (NR 10-30), TIBC 43.8 μm (NR 45-70), iron saturation 9%. Normal results included urea, creatinine, serum B12, red cell and serum folate, immunoglobulin concentrations and serum electrophoretic pattern. No Bence Jones protein was detected. Serology for hepatitis A and B was negative. Culture of blood, urine, and sputum yielded no pathogens. A Ziehl-Nielsen (ZN)/auramine stain of sputum was negative as was a Mantoux test. A CT scan of the abdomen showed the liver and spleen to be normal in texture; there was no lymphadenopathy. Chest radiography and a skeletal survey were normal. The peripheral blood film showed rouleaux and occasional abnormal cells. Bone marrow aspirate was difficult and yielded a poor aspirate containing cells such as are shown in Figure 1. A trephine biopsy was markedly abnormal (Figure 2). A further diagnostic procedure was performed.
Leukaemic cells from 2 patients with B-prolymphocytic leukaemia were immortalized in vitro by mea... more Leukaemic cells from 2 patients with B-prolymphocytic leukaemia were immortalized in vitro by means of Epstein-Barr virus and phorbol-ester TPA. The resulting cell lines, named JVM-2 and JVM-3, have been growing continuously in liquid culture for more than one year. JVM-2 is characterized cytogenetically by t(11;14)(q13; q32), and JVM-3 by trisomy 12. The immunoglobulin (Ig) heavy- and light-chain genes showed the same pattern of rearrangement in both lines as in the original prolymphocytes from each case. The cells from these lines showed a spectrum of morphological and immunological features corresponding to different stages of B-cell maturation. The expression of Ig, IgM-lambda in JVM-2 and IgMD-K in JVM-3, changed from a predominantly membrane pattern in the original cells to a cytoplasmic one in the cell lines. By comparison with their original progenitors, the cells from both lines showed reduced reactivity with the monoclonal antibody (MAb) FMC7, and increased expression of the antigens recognized by the MAbs OKT10, alpha-Tac, FMC53 and Ki-I. The availability of cell lines from this rare type of lymphoid leukaemia offers a potential tool for the study of molecular events associated with the expression of Ig and other antigens by neoplastic cells.
Sixteen patients with chronic granulocytic leukaemia (CGL) in blast cell transformation (BT) were... more Sixteen patients with chronic granulocytic leukaemia (CGL) in blast cell transformation (BT) were studied by means of serial bone marrow trephine biopsies (BMB). The BMB were performed during the following stages of the disease: 1 at diagnosis of BT, 2 following ablative therapy and 3 during haematological recovery following autografting. In 10 of the 16 patients, BT was recognized by the presence of a distinctive infiltration with blast cells with a single large hyperchromatic nucleolus. In four of the 16 patients, the diagnosis of BT could only be made by BMB since simultaneous marrow aspiration yielded either a dry tap or no marrow fragments; in two of these four patients the BT was focal in the BMB specimen. Following ablative therapy and during haematological regeneration after autografting, BM aspirates were unsatisfactory in most cases and they were inadequate to assess cellularity or the presence of residual blasts. In contrast, sections of BMB showed clearly whether a decrease in cellularity took place or, in some, residual blasts were still present. BMB samples obtained 2 weeks after autografting showed haemopoietic regeneration consisting of discrete foci of either erythroid, granulocytic or megakaryocytic precursor cells. We conclude that BMB is essential for diagnosing CGL in BT, for monitoring the progress of these patients after therapy and in assessing the haemopoietic regeneration following autografting.
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