acid-phosphatase and Leukemia--Hairy-Cell

Hairy cell leukaemia is a rare chronic neoplastic B-cell lymphoproliferation that characteristically involves blood, bone marrow and spleen with liver, lymph node and skin less commonly involved. Histologically, the cells have a characteristic appearance with pale/clear cytoplasm and round or reniform nuclei. In the spleen, the infiltrate involves the red pulp and is frequently associated with areas of haemorrhage (blood lakes). The cells stain for B-cell related antigens as well as with antibodies against tartrate-resistant acid phosphatase, DBA44 (CD72), CD11c, CD25, CD103, CD123, cyclin D1 and annexin A1. Mutation of BRAF -V600E is present and antibody to the mutant protein can be used as a specific marker. Bone marrow biopsy is essential in the initial assessment of disease as the bone marrow may be inaspirable or unrepresentative of degree of marrow infiltration as a result of the tumour associated fibrosis preventing aspiration of the tumour cell component. Bone marrow biopsy is important in the assessment of therapy response but in this context staining for CD11c and Annexin A1 is not helpful as they are also markers of myeloid lineage and identification of low level infiltration may be obscured. In this context staining for CD20 may be used in conjunction with morphological assessment and staining of serial sections for cyclin D1 and DBA44 to identify subtle residual infiltration. Staining for CD79a and CD19 is not recommended as these antibodies will identify plasma cells and can lead to over-estimation of disease. Staining for CD20 should not be used in patients following with anti-CD20 based treatments. Down regulation of cyclin D1 and CD25 has been reported in patients following BRAF inhibitor therapy and assessment of these antigens should not be used in this context. Histologically, hairy cell leukaemia needs to be distinguished from other B-cell lymphoproliferations associated with splenomegaly including splenic marginal zone lymphoma, splenic diffuse red pulp small B-cell lymphoma and hairy cell leukaemia variant. This can be done by assessment of the spleen but as this is now rarely performed in this disorder distinction is almost always possible by a combination of morphological and immunophenotypic studies on bone marrow trephine biopsy, which can be supplemented by assessment of BRAF-V600E mutation assessment in borderline cases.

Topics: Acid Phosphatase; Antigens, CD; B-Lymphocytes; Bone Marrow; Cyclin D1; Diagnosis, Differential; Gene Expression; Humans; Immunohistochemistry; Isoenzymes; Leukemia, Hairy Cell; Liver; Lymph Nodes; Lymphoma, B-Cell, Marginal Zone; Proto-Oncogene Proteins B-raf; Skin; Spleen; Tartrate-Resistant Acid Phosphatase

Hairy cell leukemia (HCL) is characterized by leukemic cells with abundant "hairy" cytoplasm, strong cytoplasmic positivity for tartrate-resistant acid phosphatase (TRAP), characteristic immunophenotype and sensitivity to treatment with purine nucleoside analogs. HCL-variant (HCL-v) encompasses chronic B-cell leukemias resembling classical HCL but exhibiting variant cytomorphology, variant immunophenotype and resistance to conventional HCL therapy. We present the case of a 67-year-old Taiwanese male with HCL-v who had leukocytosis and splenomegaly. His hairy leukemic cells were weakly positive for TRAP and expressed CDllc and CD103 but not CD25. He received oral chemotherapy with chlorambucil and in complete hematological remission in 9 months but relapsed 2 months later. Literature review revealed 9 cases of HCL and 3 cases of HCL-v including current case from Taiwan. All patients were adults with splenomegaly. The HCL patients had a significantly higher frequency of leukopenia (p = 0.024) and monocytopenia (p = 0.008) and a lower frequency of leukocytosis (p = 0.018) than HCL-v patients. All 8 HCL patients responded favorably to 2-chlorodeoxyadenosine with or without splenectomy. The 3 HCL-v patients had leukocytosis and received chemotherapy with variable outcome. HCL and HCL-v are rare in Taiwan and their pathological and immunophenotypical features were not fully characterized. A multimodality approach incorporating hematological findings, cytomorphology, histopathology, cytochemistry, complete immunophenotyping and clinical features is needed to identify and characterize such cases in Taiwan.

Topics: Acid Phosphatase; Aged; Antigens, CD; Antineoplastic Agents, Alkylating; Biomarkers, Tumor; Bone Marrow; Chlorambucil; Humans; Isoenzymes; Leukemia, Hairy Cell; Leukocytosis; Male; Recurrence; Remission Induction; Splenomegaly; Tartrate-Resistant Acid Phosphatase

Topics: Acid Phosphatase; Biomarkers; Clinical Enzyme Tests; Diagnosis, Differential; Hematologic Diseases; Hematologic Tests; Humans; Leukemia, Hairy Cell; Leukocytes; Reference Values; Specimen Handling; Tartrates

Acid phosphatases (APs) are a family of enzymes that are widespread in nature, and can be found in many animal and plant species. Mystery surrounds the precise functional role of these molecular facilitators, despite much research. Yet, paradoxically, human APs have had considerable impact as tools of clinical investigation and intervention. One particular example is tartrate resistant acid phosphatase, which is detected in the serum in raised amounts accompanying pathological bone resorption. This article seeks to explore the identity and diversity of APs, and to demonstrate the relation between APs, human disease, and clinical diagnosis.

Topics: Acid Phosphatase; alpha-Macroglobulins; Biomarkers; Bone Resorption; Favism; Gaucher Disease; Humans; Intracellular Fluid; Isoenzymes; Leukemia, Hairy Cell; Male; Osteoclasts; Osteoporosis; Prostate; Prostatic Neoplasms; Protein Binding; Reactive Oxygen Species; Tartrate-Resistant Acid Phosphatase

Topics: Acid Phosphatase; Cytogenetics; Diagnosis, Differential; Humans; Immunologic Techniques; Leukemia, Hairy Cell; Microscopy, Electron; Tartrates

Topics: Acid Phosphatase; Adult; Diagnosis, Differential; Drug Resistance; Female; Histocytochemistry; Humans; Leukemia; Leukemia, Hairy Cell; Male; Middle Aged; Tartrates

Topics: Acid Phosphatase; Adult; Anemia, Aplastic; Antibody-Dependent Cell Cytotoxicity; DNA Nucleotidylexotransferase; Female; Hematologic Diseases; Histocytochemistry; Humans; Infectious Mononucleosis; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphoma; Male; Middle Aged; Phenotype; Skin Neoplasms; T-Lymphocytes

Topics: Acid Phosphatase; Adrenal Cortex Hormones; Adult; Aged; Bone Marrow; Female; Humans; Leukemia, Hairy Cell; Liver; Lymph Nodes; Male; Mechlorethamine; Mercaptopurine; Middle Aged; Radiography; Spleen; Splenectomy

We have studied ten patients with a lymphoproliferative disorder characterized by massive splenomegaly; minimal lymphadenopathy; varying degrees of blood cytopenias; circulating atypical lymphoid cells frequently with "hairy" cytoplasm; monoclonal serum paraprotein; and tartrate-resistant acid phosphatase reactivity in the tumor cells of five patients tested. Although the clinical and laboratory features in most cases prompted a clinical diagnosis of "hairy cell leukemia" (HCL), histologic, ultrastructural and immunohistologic studies of multiple organs revealed distinctive features recognizably different from leukemic reticuloendotheliosis (LRE). Because this B lymphocyte proliferation may be mistaken for LRE in cases where careful histologic study is not performed, it may be responsible in part for the conflicting data in attempts to characterize the cell of origin of the latter disease. Clinical and experimental data in HCL must be questioned if they do not include histopathologic confirmation of the diagnosis.

Topics: Acid Phosphatase; Aged; Diagnosis, Differential; Female; Humans; Immunoglobulin M; Leukemia, Hairy Cell; Lymph Nodes; Lymphoma, Non-Hodgkin; Male; Microscopy, Electron; Middle Aged; Splenomegaly; Tartrates

A laboratory and clinical evaluation of 24 patients with hairy cell leukemia was carried out over a 23-month period. Most patients had splenomegaly without adenopathy or pancyotpenia. Nine of the patients had undergone splenectomy prior to referral; their median WBC count was 6600/mm3. The median WBC count for the 14 patients who had no prior therapy was 3550/mm3, and their median platelet count was 80,500/mm3. Spleen weights ranged from 618 to 3780 g; there appeared to be no relationship between the size of the spleen and the response in the blood counts after splenectomy. Four patients in whom the majority of the WBC were hairy cells underwent splenectomy, which produced no real change in their WBC count; however, there was improvement in the platelet count in three. In contrast, the presence of leukopenia with a low percentage of hairy cells predicted a beneficial response to splenectomy. The study of surface immunoglobulins (SIg) in 16 patients demonstrated that resynthesis had occurred in each case. Phagocytosis of zymosan was studied in 15 patients; in 8 of these, 25% or more of the hair cells were capable of phagocytosis; in 6 others, 0--9%; and in one, 13%. The resynthesis of SIg is a feature usually associated with B-lymphocytes, but the phagocytosis of zymosan is not. Thus, the existence of either a spectrum of functional capabilities of hairy cells or several distinct subtypes is suggested by these data. Platelet aggregation with epinephrine was abnormal in 7 of 14 patients studied but there were no clinically significant bleeding problems. A chromosome abnormality was present in 2 of the 19 patients from whom adequate samples were obtained; the abnormality probably involved chromsome 12 in both patients as well as absent Y and was associated with a rapidly progressive clinical course. The presence of a predominant number of hairy cells with a normal or increased peripheral blood WBC count or of a chromosomal abnormality suggests that splenectomy might not be beneficial as the initial therapy and that chemotherapy should be considered.

Topics: Acid Phosphatase; Adult; Aged; Antineoplastic Agents; Bone Marrow; Chromosome Aberrations; Drug Therapy, Combination; Female; Humans; Leukemia, Hairy Cell; Male; Middle Aged; Pancytopenia; Phagocytosis; Platelet Aggregation; Receptors, Antigen, B-Cell; Rosette Formation; Splenectomy; Zymosan

According publications' data, hairy cell leukosis encounters in humans aged from 26 to 70 years and in males four times more often than in females. The disease is manifested by impotence, splenomegaly, and presence in blood and bone marrow clone of lymphoid cells with particular morphology, cytochemic markers and immune phenotype (sIg+, CD19+, CD20+, CD5-, CD10-, with marked expression of CD25+, CD103+). The presented clinical case of illness with hairy cell leucosis is the first one detected in pediatric practice in adolescent of 16 years old.

Topics: Acid Phosphatase; Adolescent; Antigens, CD; Antigens, CD19; Antigens, CD20; B-Lymphocytes; Female; Flow Cytometry; Humans; Integrin alpha Chains; Interleukin-2 Receptor alpha Subunit; Isoenzymes; Leukemia, Hairy Cell; Lymphocytes

Annexin-1 and T-bet are recently described immunohistochemical stains that reportedly assist in the diagnosis of hairy cell leukemia (HCL). Our objective was to assess the sensitivity and specificity of a panel of immunohistochemical stains in distinguishing HCL from other B-cell neoplasms, particularly splenic and extranodal marginal zone lymphomas (SMZL and ENMZL, respectively). The study included 234 bone marrow biopsy specimens: 101 HCL, 13 SMZL, and 10 ENMZL cases were assessed with CD20, tartrate-resistant acid phosphatase (TRAP), DBA.44, a-1, T-bet, and cyclin D1, and 110 control cases were assessed with annexin-1 and T-bet. Our study showed that annexin-1 is a specific and sensitive marker for HCL; however, interpretation is limited by positivity within myeloid precursors. T-bet, DBA.44, and TRAP immunohistochemical stains lack sufficient sensitivity and specificity to differentiate HCL from either form of marginal zone lymphoma. However, our data suggest that the addition cyclin D1 to the immunostaining panel will increase the sensitivity and specificity of HCL diagnosis.

Topics: Acid Phosphatase; Annexins; Antigens, CD20; Bone Marrow; Cyclin D1; Diagnosis, Differential; Humans; Immunohistochemistry; Isoenzymes; Leukemia, Hairy Cell; Lymphoma, B-Cell; Sensitivity and Specificity; T-Box Domain Proteins; Tartrate-Resistant Acid Phosphatase

Topics: Acid Phosphatase; Diagnosis, Differential; Humans; Isoenzymes; Leukemia, Hairy Cell; Leukocyte Count; Mutation; Proto-Oncogene Proteins B-raf; Tartrate-Resistant Acid Phosphatase

The differential diagnosis of hairy cell leukemia (HCL) includes HC variant (HC-V) and marginal zone lymphoma (MZL). There is a high sensitivity of combined DBA.44/TRAP-positivity (+) in confirming HCL. A previous study of mucosa-associated lymphoid tissue lymphoma showed DBA.44+ in 10%, TRAP+ in 29%, and DBA.44+/TRAP+ in 5%.. We now study nodal MZL (NMZL) and HC-V.. Two HCL, 2 HC-V, 3 MZL of bone marrow (BM), 2 MZL versus B-cell lymphoma, not otherwise specified (BCL, NOS) of BM, and 4 NMZL and 5 extranodal MZL (EMZL) were stained with DBA.44 and TRAP and reviewed for staining pattern/intensity.. DBA.44 intensely stained all cells in 2/2 HCL, 1/2 HC-V, and 1 EMZL; intensely stained >20% of neoplastic cells (NCs) in 7 MZLs (1 BM, 3 NMZL, and 3 EMZL); and was negative/stained <10% of NCs in 1/2 HC-V, the remaining MZLs (2 BM, 1 NMZL, and 1 EMZL), and 2/2 MZL versus BCL, NOS-BM. TRAP intensely stained all cells in 2/2 HCL, the DBA.44+ HC-V, and 1 MZL versus BCL, NOS-BM; intensely stained >20% of NCs in 1 MZL versus BCL, NOS-BM, 1 MZL-BM, and 1 EMZL; and was negative in the remainder (1 HC-V, 2 MZL-BM, 1 MZL vs. BCL, NOS-BM, the 4 NMZL, 3 EMZL) in which it was able to be performed. There was combined DBA.44+/TRAP+ in 2/2 HCL, 1/2 HCV, 1/3 MZL-BM, and 1/5 EMZL. Only 1 case (MZL vs. BCL, NOS-BM) was TRAP+/DBA.44-.. Although combined intense, diffuse TRAP+/DBA.44+ is highly sensitive for HCL, it is not entirely specific, and may be observed in HC-V and EMZL, further supporting a histogenetic relationship between these entities.

Topics: Acid Phosphatase; Biomarkers, Tumor; Diagnosis, Differential; Humans; Isoenzymes; Leukemia, Hairy Cell; Lymphoma, B-Cell; Ribosomal Proteins; Tartrate-Resistant Acid Phosphatase

Because of marrow fibrosis, bone marrow aspirations are often nonconclusive in patients with hairy cell leukemia (HCL). Therefore, histologic examination is important in HCL but often difficult in cases with low numbers of tumor cells. A combined immunohistochemical positivity for DBA.44 and tartrate-resistant phosphatase was previously found in 100% of HCL and suggested to be specific for this diagnosis. To further assess the diagnostic specificity and sensitivity of this immunohistochemical approach in a higher number of cases, we analyzed 56 HCLs and lymphoma tissue microarrays, including 840 cases of the most frequent non-Hodgkin lymphomas. All HCLs showed combined positivity for these two proteins (100% sensitivity). Both antibodies were often positive in other lymphoma types. DBA.44 reactivity was especially frequent in follicular lymphomas (46%), whereas tartrate-resistant acid phosphatase (TRAP) expression was often seen in mantle cell lymphomas (57%), primary mediastinal large B-cell lymphomas (54%), and chronic lymphocytic leukemia/small lymphocytic lymphoma (41%). A combined DBA.44/TRAP positivity was seen in only 3% of non-HCL non-Hodgkin lymphomas, including cases of diffuse large B-cell lymphomas, follicular lymphomas, chronic lymphatic leukemia/small lymphocytic leukemias, and mantle cell lymphomas. Overall, these data confirm the utility of combined immunohistochemical DBA.44/TRAP expression analysis in confirming the diagnosis of HCL. However, combined positivity for these markers is highly sensitive but not absolutely specific for HCL.

Topics: Acid Phosphatase; Antibodies, Monoclonal; Humans; Immunoenzyme Techniques; Isoenzymes; Leukemia, Hairy Cell; Lymphoma, Non-Hodgkin; Sensitivity and Specificity; Tartrate-Resistant Acid Phosphatase; Tissue Array Analysis

Hairy cell leukemia (HCL) is a rare chronic B-cell lymphoproliferative disorder characterized by splenomegaly, pancytopenia, and circulating atypical lymphocytes with circumferential cytoplasmic projections. We investigated the specificity and the sensitivity of anti-TRAP antibody immunoreactivity in 57 cases of HCL. We found that there is a statistically highly significant difference between TRAP immunoreactivities of the study and the control groups, and HCL can be diagnosed by TRAP immunoreactivity in bone marrow trephine biopsy materials with a specificity of 98.27 % and a sensitivity of 100%.

Topics: Acid Phosphatase; Antibodies, Monoclonal; Antigens, CD20; Bone Marrow Cells; CD5 Antigens; Humans; Immunohistochemistry; Isoenzymes; Leukemia, Hairy Cell; Liver; Osteoclasts; Spleen; Tartrate-Resistant Acid Phosphatase

We have previously reported that bryostation 1 (Bryo 1) induces differentiation of chronic lymphocytic leukemia (CLL) in vitro to a hairy cell (HC) stage. This study tests the hypothesis that Bryo 1-differentiated CLL cells are more susceptible to 2-chlorodeoxyadenosine (2-CdA) than parent CLL cells. A recently established EBV-negative CLL line (WSU-CLL) from a patient resistant to chemotherapy including fludarabine was used to test this hypothesis. Both Bryo 1 (10-1000 nM) and 2-CdA (5.6-22.4 microM) exhibited a dose-dependent growth inhibitory effect on the WSU-CLL cell line. In vitro, the sequential exposure to Bryo 1 (100 nM for 72 h) followed by 2-CdA (11.2 microM) resulted in significantly higher rates of growth inhibition than either agent alone. Changes in immunophenotype, enzymes, lipids, proteins, and the DNA of WSU-CLL cells were studied before and after Bryo 1 treatment. Bryo 1 induced a positive tartrate-resistant acid phosphatase reaction and two important markers, CD11c and CD25, after 72 h of culture, confirming the differentiation of CLL to HC. The Fourier transformation infrared spectroscopic analysis showed that the amount of membrane lipids significantly increased in Bryo 1-treated cells compared to controls after 24 h, whereas the protein content, as well as the DNA content, decreased. This finding supports the change of CLL to HC. To evaluate the in vivo efficacy of Bryo 1 and 2-CdA, we used a xenograft model of CLL in WSU-CLL-bearing mice with severe combined immune deficiency. s.c. tumors were developed by injection of 10(7) WSU-CLL cells, and fragments were then transplanted into a new batch of severe combined immunodeficient mice. Bryo 1 and 2-CdA at the maximum tolerated doses (75 micrograms/kg i.p. and 30 mg/kg s.c., respectively) were administered to the mice at different combinations and schedules. The survival in days, the tumor growth inhibition ratio, the tumor growth delay, and the log10 kill of the mice treated with Bryo 1 followed by 2-CdA were significantly better than the control and other groups. We conclude that the sequential treatment with Bryo 1 followed by 2-CdA resulted in higher antitumor activity and improved animal survival.

Topics: Acid Phosphatase; Aged; Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Bryostatins; Cell Division; Cladribine; DNA, Neoplasm; Drug Administration Schedule; Drug Screening Assays, Antitumor; Humans; Lactones; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Lipid Metabolism; Macrolides; Male; Mice; Mice, SCID; Neoplasm Proteins; Neoplasm Transplantation; Spectroscopy, Fourier Transform Infrared; Transplantation, Heterologous; Tumor Cells, Cultured

Tartrate-resistant acid phosphatase (TRAP) is expressed abundantly by osteoclasts and is required for bone resorption. This enzyme is emerging as an important biomarker in bone pathology, both for histochemical identification of osteoclasts and as a serum marker of osteoclast activity and increased bone turnover. Rat and mouse models are becoming popular systems for studying osteoclast development, bone physiology and morphogenesis, and bone diseases such as osteoporosis. We have developed two unique antibodies to human TRAP purified from hairy cell leukemia spleen. Both antibodies (9C5 and 14G6) are suitable for immunohistochemistry of osteoclasts and macrophages. Only one (14G6) is capable of immunoprecipitating active TRAP from human cell lysates. Antibody 9C5 reacts with a denatured epitope of TRAP while antibody 14G6 probably reacts with a native, conformational determinant. The high degree of homology among TRAPs of various species predicts that these antibodies should be suitable for work in experimental animals as well as humans. Immunohistochemical staining, electrophoretic analyses, immunoprecipitation and immunoblotting assays of human rat and mouse TRAP were carried out to test the validity of these antibodies as cell markers in rodents. Both antibodies were suitable for immunohistochemistry in all species. Antibody 9C5 was suitable for immunoblotting of denatured TRAP of all species tested. Antibody 14G6 reacted with the native TRAP of humans only and failed to immunoprecipitate mouse or rat TRAP activity. Although TRAP is a phylogenetically conserved protein, subtle, species-specific determinants exist. Care should be exercised when anti-TRAP antibodies are used for immunoassay in experimental animals.

Topics: Acid Phosphatase; Animals; Antibodies, Monoclonal; Biomarkers; Epitope Mapping; Humans; Immunohistochemistry; Isoenzymes; Leukemia, Hairy Cell; Mice; Rats; Species Specificity; Tartrate-Resistant Acid Phosphatase; Tumor Cells, Cultured

A major product of osteoclasts, tartrate-resistant acid phosphatase (TRAP) is an essential but insufficient enzyme for bone resorption. TRAP is an excellent cell marker for osteoclasts and macrophages and is being investigated as a serum marker for osteoclast activity in diseases of bone destruction. For decades, TRAP has also been used as a marker for hairy cell leukemia. Immunoassays for TRAP are sought to increase the sensitivity and specificity of the TRAP test for bone and hairy cells. Our laboratory recently developed a monoclonal antibody to TRAP (9C5) useful for immunohistochemical identification of TRAP-positive cells in paraffin sections. Herein, we characterize 9C5 in greater detail and report production of another anti-TRAP monoclonal antibody antibody (14G6) reactive with native, active enzyme antigen. Enzyme immunoassay, immunoprecipitation, western blot, and immunohistochemical analyses revealed the contrasting properties of 9C5 and 14G6. Antibody 9C5 reacts with a heat-denatured epitope and is suitable for denaturing western blot analysis and for immunohistochemistry. Antibody 14G6 reacts with a conformational determinant destroyed by heat and is suitable for immunoprecipitation of active TRAP, although 20% to 30% of activity is inhibited in the immune complexes. Having characterized several properties of these anti-TRAP antibodies, 9C5 and 14G6 may be useful for development of TRAP-specific immunoassays in bone pathology and hematology. They will certainly be of use for the study of biosynthesis, regulation, expression, and function of TRAP.

Topics: Acid Phosphatase; Antibodies, Monoclonal; Antibody Specificity; Biomarkers; Blotting, Western; Bone Resorption; Epitopes; Humans; Hybridomas; Immunoenzyme Techniques; Immunohistochemistry; Isoenzymes; Leukemia, Hairy Cell; Macrophages; Monocytes; Tartrate-Resistant Acid Phosphatase

A cell line, JHC-2, was established from the peripheral blood of a patient with hairy cell leukemia (HCL)-Japanese variant. The JHC-2 cells have cytologic features similar to those of the original tumor cells. They displayed hairy cytoplasmic projections by phase contrast and scanning electron microscopy. The tartrate-resistant acid phosphatase reaction was weakly positive. The immunophenotype of the JHC-2 cells was CD5-, CD10-, CD11c+/-, CD19+, CD21+, CD23+, CD24-, CD25+/-, CD38- and FMC-7+. The expression of surface immunoglobulin (IgG, kappa) and the configuration of Ig gene rearrangements in the JHC-2 cells were identical to those in the original leukemic cells, and the JHC-2 cells displayed trisomy 9 on cytogenetic examination. Southern blot analysis for the Epstein-Barr virus (EBV) genome showed that the JHC-2 cells contained the EBV genome, although the freshly isolated leukemic cells did not. These results indicate that the JHC-2 cell line is an EBV spontaneously transformed B cell line originating from HCL cells.

Topics: Acid Phosphatase; Antigens, CD; Antigens, Surface; Gene Rearrangement; Herpesvirus 4, Human; Humans; Immunoglobulin G; Immunoglobulin kappa-Chains; Isoenzymes; Japan; Leukemia, Hairy Cell; Male; Middle Aged; Phenotype; Receptors, Interleukin-2; Tartrate-Resistant Acid Phosphatase; Tumor Cells, Cultured

The demonstration of tartrate-resistant acid phosphatase (TRAP) activity has long been a cornerstone in the diagnosis of hairy cell leukemia (HCL). Recently a monoclonal antibody to this enzyme has been developed that can be used in an immunoperoxidase method on paraffin-embedded tissues. By using a peroxidase-labeled streptavidin biotin method, paraffin sections of B5 and formalin-fixed tissue from 86 cases of HCL (41 bone marrow, 36 spleen, 9 liver) were stained with the antibody to TRAP and compared against staining for CD20 (L26) and DBA.44 (DAKO, Carpinteria, Calif). In addition, 193 specimens (127 bone marrow, 42 lymph node, 19 spleen, 5 other) from a variety of neoplastic and nonneoplastic hematologic conditions were stained using the monoclonal antibody to TRAP. For comparison, these cases were also stained with DBA.44. In the cases of HCL, 80 of 86 specimens were immunoreactive for TRAP. While the antibody to TRAP generally stained less than 50% of the hairy cells, CD20 and DBA.44 stained 90% and 50% to 60% of hairy cells, respectively. Two of three cases of marginal zone lymphoma showed weak immunoreactivity to the TRAP antibody. Two specimens from a patient with Gaucher's disease and 8 of 13 cases of mastocytosis also showed positivity to the TRAP antibody in the macrophages and mast cells, respectively. In contrast, staining for DBA.44 was positive in 3 of 9 cases of B-cell large cell lymphoma, 1 of 4 cases of mantle cell lymphoma, and in the paraimmunoblasts of 1 of 7 cases of small lymphocytic lymphoma. Only HCL was TRAP and DBA.44 positive. This antibody to TRAP is a useful addition to the diagnosis of HCL but should be used in conjunction with CD20 and DBA.44. The use of this antibody to determine minimal residual disease after chemotherapy was not addressed.

Topics: Acid Phosphatase; Antibodies, Monoclonal; Biomarkers, Tumor; Bone Marrow; Bone Marrow Neoplasms; Diagnosis, Differential; Gaucher Disease; Humans; Immunohistochemistry; Isoenzymes; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Liver; Liver Neoplasms; Lymph Nodes; Lymphoma, B-Cell; Lymphoproliferative Disorders; Macrophages; Mast Cells; Paraffin Embedding; Pathology, Clinical; Spleen; Splenic Neoplasms; Tartrate-Resistant Acid Phosphatase

We have developed a monoclonal antibody (9C5) for immunohistochemical localization of tartrate-resistant acid phosphatase (TRAcP). This antibody reacts with a denatured epitope of TRAcP and requires enhancement methods to promote antigenicity in paraffin-embedded tissues. We used this antibody to systematically examine proteolytic digestion and heat denaturation conditions for epitope enhancement in both paraffin sections and fixed smears. The goal was to increase the sensitivity of the immunohistochemical stain for TRAcP. Optimal conditions for proteolytic digestion were established. Denaturation in a conventional boiling water bath was compared to microwave irradiation in several commonly used solutions. Immunohistochemistry was compared directly to TRAcP cytochemistry in fixed smears from hairy cell leukemia specimens to gauge the level of sensitivity of our improved method. Attempts were made to "retrieve" the 9C5 epitope from overfixed tissues and aged smears. Maximal immunoreactivity of TRAcP was achieved by microwave irradiation in a citrate or Tris buffer of pH 6.0-8.0 without the need for a subsequent protease digestion step. With this method of epitope enhancement, immunohistochemistry with antibody 9C5 was as sensitive as direct cytochemical staining of TRAcP activity. However, once a tissue specimen had been overfixed or a smear stored for a year or more, the 9C5 epitope was no longer retrievable. The key element in epitope enhancement for 9C5 immunohistochemistry is heat denaturation of the target epitope. Immunohistochemistry of TRAcP in paraffin sections would be a great asset to the study of specialized forms of the monocyte/macrophage lineage and to the process of macrophage activation. It would also provide another means for more precise evaluation of residual disease in bone marrow of patients treated for hairy cell leukemia.

Topics: Acid Phosphatase; Biomarkers, Tumor; Epitopes; Humans; Immunohistochemistry; Isoenzymes; Leukemia, Hairy Cell; Sensitivity and Specificity; Tartrate-Resistant Acid Phosphatase

B-Chronic lymphocytic leukemia (B-CLL) and hairy cell leukemia (HCL) are both differentiated B-cell lymphoproliferative disorders. Prior studies have suggested that phorbol esters and the macrocyclic lactone Bryostatin-1, which are both protein kinase-C activators, can induce the differentiation of B-CLL cells into HCL cells in vitro, as evidenced by morphology, phenotype and TRAP activity. The differentiating effect of all-trans retinoic acid on B-CLL cells has been less extensively studied. We studied the effects of incubating adherence purified B-CLL cells with phorbol myristic acetate (PMA), all-trans retinoic acid (ATRA), and Bryostatin-1. None of these agents induced a true HCL phenotype (CD5-, CD11c/CD25 coexpression) under the conditions studied.

Topics: Acid Phosphatase; Antigens, CD; Antigens, Neoplasm; B-Lymphocytes; Biomarkers, Tumor; Bryostatins; Cell Differentiation; Enzyme Activation; Humans; Immunophenotyping; Isoenzymes; Lactones; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Macrolides; Neoplastic Stem Cells; Protein Kinase C; Tartrate-Resistant Acid Phosphatase; Tetradecanoylphorbol Acetate; Tretinoin; Tumor Cells, Cultured

Hairy cell leukemia (HCL) is an uncommon type of chronic B cell leukemia mainly affecting middle-aged adults. HCL presenting with pancytopenia is rare in Japan and a distinct subtype of HCL termed HCL-Japanese variant is predominantly seen. We describe a HCL patient with unusual presentation. The patient was a 26-year-old male, such early onset of HCL being quite rare. The patient showed leukocytosis with many circulating hairy cells and cellular bone marrow. These findings were preferentially seen in HCL-Japanese variant, but, cytomorphologic, cytochemical and immunophenotypical studies on the pathologic cells were consistent with those of typical HCL seen in Western countries. Interferon-alpha therapy was very effective in this case. Differentiation of the subtype of HCL appears to be important for the choice of the treatment. The cytological findings were useful for the differential diagnosis of HCL presenting with leukocytosis.

Topics: Acid Phosphatase; Adult; Age of Onset; Biomarkers, Tumor; Diagnosis, Differential; Humans; Interferon-alpha; Isoenzymes; Leukemia, Hairy Cell; Leukocytosis; Male; Receptors, Interleukin-2; Splenomegaly; Tartrate-Resistant Acid Phosphatase

Immunohistochemical studies were done on formalin-fixed, paraffin-embedded tissues to evaluate the specificity of a newly developed monoclonal antibody (9C5) against tartrate-resistant acid phosphatase. Sections from 195 specimens were examined, which included 33 types of tissues/organs. These tissues included normal, inflammatory, and neoplastic processes. Neoplastic tissues from 14 patients with hairy cell leukemia served as positive controls. Epitope enhancement was accomplished either by microwave irradiation in citrate buffer or by boiling in water followed by trypsin digestion. Tissues were reacted with monoclonal antibody 9C5 and stained with either the avidin-biotin peroxidase method or the alkaline phosphatase anti-alkaline phosphatase method. The hairy cells of all cases of hairy cell leukemia reacted positively with 9C5. Other positively stained cells included osteoclasts, activated macrophages and giant cells. Immunohistochemical studies with 9C5, when interpreted within the context of the specificity of this antibody, are useful for the diagnosis and assessment of treatment results for hairy cell leukemia. Monoclonal antibody 9C5 also may be useful as a marker for osteoclasts and the activated macrophages and for the diagnosis of disorders involved by these cells.

Topics: Acid Phosphatase; Antibodies, Monoclonal; Antibody Specificity; Biomarkers, Tumor; Hematopoiesis; Humans; Immunohistochemistry; Inflammation; Isoenzymes; Kupffer Cells; Leukemia, Hairy Cell; Macrophages; Neoplasms; Reference Values; Tartrate-Resistant Acid Phosphatase

Tartrate-resistant acid phosphatase (TRAcP) has been an indispensible marker for hairy cell leukemia (HCL) for over two decades. However, the traditional TRAcP cytochemical stain cannot be performed effectively on sections of paraffin-embedded tissues that are important resources for histopathologic evaluation in diagnosis and treatment of HCL. Wide variation in expression of TRAcP activity by hairy cells (HCs) within and among patients is an interesting biologic phenomenon that has not been explained and can cause some diagnostic uncertainty as well. To solve the problem of staining TRAcP in paraffin sections and to begin to address the questions of variable TRAcP expression in HCL, we developed a monoclonal antibody to TRAcP, 9C5, for immunohistochemical identification of HCs. In smears of blood and bone marrow, immunocytochemistry of TRAcP using 9C5 was as specific but slightly less sensitive than direct cytochemical staining of enzymatic activity. In paraffin sections of spleen and bone marrow from HCL patients, immunohistochemistry with 9C5 stained the HCs with high sensitivity and specificity and clearly showed the characteristic diffuse infiltration by HCs. Other cells noted to stain strongly with 9C5 were occasional macrophages in bone marrow smears and osteoclasts and occasional tissue macrophages in paraffin sections. These are cells known to express abundant TRAcP activity. Immunohistochemistry with anti-TRAcP monoclonal antibody 9C5 may have utility as an added option in the diagnosis of HCL, as a means to evaluate residual disease in HCL patients undergoing new treatments, and as a way to address questions regarding variable expression of TRAcP activity by HCs within and among patients with HCL. Also, 9C5 has potential as a reagent for the immunoassay of bone-derived serum TRAcP in patients with certain bone diseases and cancers with bone metastasis.

Topics: Acid Phosphatase; Antibodies, Monoclonal; Biomarkers, Tumor; Humans; Immunohistochemistry; Isoenzymes; Leukemia, Hairy Cell; Tartrate-Resistant Acid Phosphatase

We investigated the effects of stromal cells, obtained from lymph nodes with various lymphoid disorders and characterized immunocytochemically as fibroblasts, on the maturation of a cell line "HN" established from an atypical PLL (prolymphocytic leukemia) having phenotypic characteristics of both CLL (chronic lymphocytic leukemia) and HCL (hairy cell leukemia). Coculture with stromal cells, irrespective of the kind of lymphoid disease affecting the lymph node, induced a plasmacytoid cytology in HN cells, an increase of cIg gamma, and decreases of sIg gamma, CD5, CD21, HC2, and B-ly7. In contrast, coculture with the stromal cells produced no marked effects on hairy cells from two HCL patients. Similarly, coculture with stromal cell supernatant or with various cytokines shown to be produced by bone marrow stromal cells produced no effects on HN cells. We propose that lymph node stromal cells play an important role in the differentiation of B cells through direct contact and that HN cells will be useful for investigating the differentiation pathway of chronic B cell leukemia cells.

Topics: Acid Phosphatase; Antigens, CD; B-Lymphocytes; Blood Coagulation Factors; Cell Differentiation; Cells, Cultured; Cytokines; Humans; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Leukemia, Prolymphocytic; Lymph Nodes; Lymphatic Diseases; Phenotype; Receptors, Antigen, B-Cell; Stromal Cells; Tumor Cells, Cultured

A prospective study was performed on peripheral EDTA blood samples mailed for screening purposes to a hematology laboratory. Twenty-one samples which were suspicious for hairy cell leukemia (H 6000, Technicon Co. or STKS, Coulter Co.), were evaluated using three diagnostic tests: Microscopic morphology (Giemsa stain); Microscopic controlled inhibition of lymphocyte acid phosphatase by tartrate; Flow-cytometric immunophenotyping with monoclonal antibodies (Epics Profile, Coulter Co.). A characteristic immunophenotype was found on all typical hairy cell leukemias (except two) by positivity of CD11c, CD19, CD22, and CD25. The above-mentioned immunophenotype on peripheral lymphocytes, which can moreover be supported by demonstrating positivity for FMC7, is diagnostic for hairy cell leukemia. It therefore represents a rather straightforward tool to the bone-marrow diagnosis.

Topics: Acid Phosphatase; Adult; Aged; Antibodies, Monoclonal; Antigens, CD; Female; Flow Cytometry; Humans; Immunophenotyping; Leukemia, Hairy Cell; Lymphocytes; Male; Middle Aged; Prospective Studies; Staining and Labeling

The phorbol esters induce differentiation of chronic lymphocytic leukemia (CLL) cells. Clinical use of this observation has been hampered by the fact that phorbol esters are also tumor promoters. In this study we demonstrate that another protein kinase C activator, without tumor promoting activity, has similar effects on CLL cells. Fresh leukemic cells from the peripheral blood of 13 patients with CLL were isolated and cultured in the absence (control) or presence of Bryostatin 1 or 12-0-tetradecanoylphorbol 13-acetate (TPA). Aliquots of cells were then analyzed after 24, 72 and 120 hours for morphological changes, acid phosphatase (ACP) and the co-expression of two hairy cell-associated surface antigens, CD22 and CD11c, by flow cytometry. Bryostatin 1 induced changes in shape and morphology similar to TPA, with adherence and increase in cell size, abundant cytoplasm and irregular cytoplasmic membrane. Both agents induced a statistically significant increase in the expression of CD22 and CD11c compared with control (p < 0.0008). There was no significant difference between the two agents in the degree of expression of these two markers. Both agents also induced ACP that was tartrate resistant (TRAP). These changes indicate that Bryostatin is as effective as TPA in inducing further differentiation of CLL cells to a hairy cell stage.

Topics: Acid Phosphatase; Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation, B-Lymphocyte; Bryostatins; CD11 Antigens; Cell Adhesion Molecules; Cell Differentiation; Enzyme Activation; Female; Humans; Immunophenotyping; Lactones; Lectins; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Macrolides; Male; Middle Aged; Protein Kinase C; Sialic Acid Binding Ig-like Lectin 2; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured

Forty Japanese patients with hairy cell leukemia (HCL) were reviewed. Nine cases were diagnosed as typical HCL, and two cases had the features of HCL variant (prolymphocytic variant). The remaining 29 cases (72.5%) differed morphologically and hematologically from the other two groups in that they usually had a moderately high leukocyte count (average 27.9 x 10(3)/microliters), and abnormal cells showing a densely stained round nucleus and an inconspicuous nucleolus. Tartrate-resistant acid phosphatase reaction was weak, and their cells exhibited generally smooth or slightly irregular, cellular outlines in smears. The cells showed weak expression of surface immunoglobulin G (IgG) with kappa-chain predominance. CD25 antigen was not detected. Some of these findings resemble those of B-cell chronic lymphocytic leukemia, but the patients also had several important features of HCL. They had splenomegaly without significant lymphadenopathy. The abnormal cells were CD20+, CD11c+ and showed typical 'hairy morphology' under phase-contrast and scanning electron microscopy. Furthermore, spleen sections revealed diffuse infiltration by the abnormal cells in the red pulp. From these findings, we speculated that this group of patients constitute a distinct subtype of HCL which is commonly seen in Japan. We propose to term the disease as HCL Japanese variant.

Topics: Acid Phosphatase; Adult; Aged; Aged, 80 and over; Antigens, Surface; Drug Resistance; Female; Humans; Japan; Leukemia, Hairy Cell; Male; Microscopy, Electron, Scanning; Middle Aged; Tartrates

Leukemic cells from eight patients with chronic lymphocytic leukemia were isolated and cultured in the continuous presence of 12-O-tetradecanoylphorbol 13-acetate (TPA) at a concentration of 1.6 x 10(-9) M for 4-10 days. Aliquots of cells were then analyzed at intervals of 24-72 hr for changes in morphology, acid phosphatase staining (AP), and expression of two hairy cell-associated surface antigens, HCL1 (CD22, Leu 14) and HCL3 (CD11c, Leu M5). All cases studied showed typical B-CLL phenotype, and only a small proportion of cells expressed CD22 and CD11c (mean 7% and 4.9%, respectively). TPA treatment induced the coexpression of CD22 (mean 49%) and CD11c (mean 48%) and tartrate-resistant acid phosphatase in seven of eight cases. Morphologically, cells in TPA cultures expressed hairy cell features that were evident in light and electron microscopic studies. Collectively these changes indicate that TPA can induce hairy cell features on CLL cells in vitro, suggesting the later maturational stage of HCL compared with CLL.

Topics: Acid Phosphatase; Aged; Antigens, CD; Antigens, Differentiation, B-Lymphocyte; CD11 Antigens; Cell Adhesion Molecules; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Female; Humans; Lectins; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Lymphocytes; Male; Microscopy, Electron; Middle Aged; Phenotype; Phorbol Esters; Precipitin Tests; Sialic Acid Binding Ig-like Lectin 2; Tetradecanoylphorbol Acetate; Time Factors

The human nonerythrocytic acid phosphatases (AcP) are composed of seven distinct activity bands in nondenaturing polyacrylamide gel electrophoresis (PAGE) when stained using either 1-naphthyl phosphate or naphthol ASBI phosphate as substrate. They are numbered 0, 1, 2, 3, 3b, 4, and 5 according to their increasing mobility toward the cathode in acidic conditions. Of these, only the most cationic "band 5" is tartrate resistant (TRAcP). When naphthol ASBI phosphate is used as substrate, AcP activity can also be stained in situ. In the presence of tartrate, activity remains strong in the hairy cells (HC) of hairy cell leukemia (HCL). Thus, the TRAcP stain has remained a reliable marker for HC. To investigate the function of TRAcP in HC, we purified two isoforms of TRAcP from HCL spleen tissue and found them to have similar substrate specificities and inhibitor sensitivities. In this report, we describe in detail the methods for TRAcP purification and compare some of the structural properties of the two isoforms to reinforce the concept that human TRAcP is a heterogeneous group of related enzymes. Band 5 represented only 15-20% of the total TRAcP extracted from HCL spleen. The remaining 80% of TRAcP hydrolyzed p-nitrophenyl phosphate but not naphthol ASBI phosphate and was not detectable in acidic, nondenaturing PAGE gels. Band 5 was solubilized from tissue using 500 mmol/L NaCl after previous extraction with 0.5% (v/v) NP-40 removed most other AcP and TRAcP activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acid Phosphatase; Amino Acid Sequence; Biomarkers, Tumor; Colorimetry; Electrophoresis, Polyacrylamide Gel; Humans; Isoenzymes; Leukemia, Hairy Cell; Molecular Sequence Data; Molecular Weight; Species Specificity; Spleen; Tartrate-Resistant Acid Phosphatase; Tartrates

Tartrate-resistant acid phosphatase (TRAcP) is a reliable cytochemical marker for the diagnosis of hairy cell leukemia (HCL). The enzyme has been the subject of much biochemical investigation yet its function in the hairy cells (HC) is still unknown. Two TRAcPs have been purified from HCL spleen tissues by a series of chromatographic separations. The two enzymes, provisionally called peak 1 and peak 2, had specific activities of greater than 600 U/mg and 800 U/mg respectively when p-nitrophenyl phosphate (p-NPP) was used as substrate and had Km values in the range of 1 to 5 mM p-NPP. The two TRAcPs had the same substrate specificities and inhibitor sensitivities, therefore could be isoforms of the same enzyme. Their pH optima were between 5 and 6 for all substrates tested including the phosphotyrosine-containing peptide, Raytide, which was still hydrolyzed efficiently at neutral pH. Neither phosphoserine nor phosphoserine-containing casein were hydrolyzed by either enzyme. The TRAcPs of HC may thus be capable of functioning as protein-tyrosine phosphatases (PTP). High activity of a PTP could regulate the activities of protein-tyrosine kinases and thereby influence the growth and differentiation of the hairy cells.

Topics: Acid Phosphatase; Electrophoresis, Polyacrylamide Gel; Humans; Hydrogen-Ion Concentration; Hydrolysis; Isoenzymes; Leukemia, Hairy Cell; Protein Tyrosine Phosphatases; Spleen; Substrate Specificity; Tartrate-Resistant Acid Phosphatase; Tartrates

1. Osteoclasts and hairy cell leukemia spleen both contain large amounts of a band 5-tartrate-resistant acid phosphatase (TrACP). 2. We have recently purified to homogeneity a band 5 TrACP from human osteoclastomas and two isoforms of band 5 TrACP (5a and 5b) from the spleen of a patient with hairy cell leukemia. 3. Although the N-terminal amino acid sequences and the apparent molecular weights of the osteoclastoma, hairy cell leukemia spleen TrACPs were identical, there were several differences in the physical and biochemical properties between the three isoenzymes. 4. Based on these findings, it is concluded that these isoenzymes are different enzymes, but that they could have originated from a similar ancestral gene. 5. It is proposed that the osteoclastoma and hairy cell leukemia band 5 TrACPs are members of a multigene family.

Topics: Acid Phosphatase; Amino Acid Sequence; Amino Acids; Enzyme Activation; Enzyme Stability; Giant Cell Tumors; Humans; Isoenzymes; Leukemia, Hairy Cell; Molecular Sequence Data; Multigene Family; Spleen; Tartrates

Chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HCL) are two common chronic lymphoproliferative disorders, each having characteristic clinical, morphologic, and immunologic features. Phenotypically, CD5 reactivity in CLL and CD11c (Leu-M5) reactivity in HCL have characterized these two leukemias among B-cell disorders. In this study, we report 14 cases of a novel chronic lymphoproliferative disorder characterized by lymphocytosis and CD11c expression, but morphologically similar to CLL. The patients' ages ranged from 46 to 81 years (median 62). Eleven had palpable splenomegaly, five with markedly enlarged spleens; only one patient had generalized lymphadenopathy. The white blood cell count ranged from 5.2 to 131.0 x 10(9)/L (median 20.8). The morphologic diagnosis in all cases was CLL, with the cells usually having abundant cytoplasm. No morphologic features, of hairy cells were evident; tartrate-resistant acid phosphatase cytochemistry was negative in all cases. Bone marrow biopsies were available in 8 of 14. Four showed focal nodular infiltrates and two had diffuse infiltrates similar to CLL; two showed only minimal interstitial involvement. All cases expressed multiple B-cell markers, and 12 of 14 had monoclonal surface immunoglobulin. The leukemic cells of all cases strongly expressed CD11c, while CD5 was expressed in 7 of 14; only 1 of the 14 cases expressed the lymph node homing receptor, Leu-8. This unique group of leukemias appears to represent the malignant transformation of lymphocytes arising from a stage of lymphocyte differentiation between that found in typical cases of CLL and that of HCL. CD11c is known to have an important function in cellular adhesion and may be important in determining the pattern of lymphocyte tissue distribution found in this group of patients.

Topics: Acid Phosphatase; Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation; Antineoplastic Agents; B-Lymphocytes; Bone Marrow; CD11 Antigens; CD5 Antigens; Female; Histocytochemistry; Humans; Immunophenotyping; Leukemia, Hairy Cell; Leukemia, Lymphocytic, Chronic, B-Cell; Leukocytosis; Male; Middle Aged; Splenomegaly; Thrombocytopenia

The immunophenotype of hairy cell leukemia (HCL) was investigated using 20 routinely fixed paraffin-embedded tissue sections (12 bone marrows, six spleens, one liver, one lymph node) from 12 patients known to have this disease. A panel of antibodies was used, including anti-leukocyte common antigen (anti-LCA), B-lineage antibodies (LN2, MB2, L26), T-lineage reagents (MT1, UCHL1), monocytic (anti-cathepsin B) and myelomonocytic (anti-lysosyme, Mac 387) antibodies, and other less lineage-specific markers (anti-S-100, anti-alpha-1-antichymotrypsin (anti-alpha 1-ACT), anti-alpha 1-antitrypsin (anti-alpha 1-AT), anti-vimentin). Anti-LCA stained hairy cells in seven of the 12 bone marrows and consistently recognized hairy cells in the spleen, liver, and lymph nodes. Hairy cells generally reacted with B-lineage antibodies and were not labeled by T-lineage markers. No reactivity was noted with myelomonocytic antibodies, anti-S-100, anti-alpha 1-ACT, or anti-alpha 1-AT. Vimentin was expressed in the majority of cases. Tartrate-resistant acid phosphatase reactivity was demonstrated in three of the 20 routinely processed tissue sections. These data suggest that immunohistochemical studies of hairy cell leukemia in routinely processed tissue may be useful in diagnostic hematopathology and surgical pathology.

Topics: Acid Phosphatase; Humans; Immunoenzyme Techniques; Immunophenotyping; Leukemia, Hairy Cell; Paraffin

Acid phosphatase (E.C.3.1.3.2) in a giant cell bone tumor and a spleen infiltrated with hairy cells was extracted by citrate buffer and then by 0.3 mol/L NaCl. The cationic acid phosphatase in the crude extract was isolated by CM-cellulose chromatography, and further separated by high pressure liquid chromatography. The majority of the tartrate resistant acid phosphatase in the hairy cell spleen was unabsorbed on CM-cellulose and was insensitive to iron. A much larger portion of the acid phosphatase in the bone tumor, than in the spleen, was cationic and was eluted from the column by 0.8 mol/L NaCl. The cationic acid phosphatase was further separated into consecutive peaks of acid phosphatases with different sensitivity to iron. A major portion of acid phosphatase in the giant cell bone tumor was enhanced by iron, while the amounts of iron-enhanced and iron-insensitive acid phosphatase were about the same in the spleen. The differences of the phosphatases in these two types of pathologic specimens indicate the occurrence of two types of enzymes with different biological significance.

Topics: Acid Phosphatase; Bone Neoplasms; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Colorimetry; Giant Cell Tumors; Humans; Iron; Leukemia, Hairy Cell; Neoplasm Invasiveness; Spleen; Tartrates

In this report the clinical, morphologic, histologic and immunologic findings of 41 patients with hairy lymphoid cells in peripheral blood and/or bone marrow are analyzed. In 27 patients the diagnosis of hairy cell leukemia was established. 14 patients had other variants of lymphoproliferative disorders: malignant lymphoma with hairy cells--7, chronic lymphocytic leukemia with hairy cells--5, and T cell lymphoproliferative disorder with hairy cells--2 patients, respectively. Several variants of malignant lymphoma with hairy cells were defined: lymphocytic, centrocytic and lymphoplasmacytic. The importance of combined use of bone marrow biopsy and immunophenotyping for the correct diagnosis of hairy cell leukemia and other "hairy-cell" lymphoproliferative disorders is stressed. The obtained data suggest relationship between characteristic clinical manifestation (isolated splenomegaly), presence of hairy cells and CD11c-antigen expression.

Topics: Acid Phosphatase; Adult; Aged; Bone Marrow; Female; Humans; Integrin alphaXbeta2; Leukemia, Hairy Cell; Lymphoproliferative Disorders; Male; Middle Aged

Tartrate-resistant acid phosphatase type-5 was purified to apparent homogeneity from human osteoclastomas by sequential chromatography on CM-Sepharose, Phenyl-Sepharose, concanavalin A-Sepharose, FPLC Superose-12, and FPLC Mono-S. The purification over the original tissue extract was 1167-fold, with a yield of 16%. An identity in the N-terminal amino acid sequence and Mr was found between this enzyme and two type-5 tartrate-resistant acid phosphatases isolated from hairy cell leukemia spleen. However, they appeared to be different as assessed by amino acid composition. In contrast to a previous report, no evidence was found for two subunits of the tartrate-resistant acid phosphatase.

Topics: Acid Phosphatase; Amino Acid Sequence; Chromatography; Drug Resistance; Giant Cell Tumors; Humans; Leukemia, Hairy Cell; Molecular Sequence Data; Tartrates

A tartrate-resistant acid phosphatase (TrACP), which has been suggested to be very similar to the osteoclastic TrACP, was partially purified from the spleen of a patient with hairy cell leukemia. The purification procedure consisted of carboxymethyl-Sepharose, phosphocellulose, Sephacryl S-200, and phenyl-Sepharose chromatographies. Polyclonal antibodies were generated in guinea pigs with a titer of at least 1:6000. Immunohistochemical staining of fetal rat tibia with the antisera revealed that only the lysosomes of osteoclasts, but not osteoblasts, were stained. An enzyme-linked immunosorbent assay (ELISA) was developed with the antisera. There was no cross-reactivity with 1) partially purified acid phosphatases (ACPs) from normal human and beef spleens, 2) ACPs in extracts of human osteoblastic cells, 3) purified bovine bone matrix TrACP, or 4) commercial prostatic ACP. However, extracts of giant cell bone tumors, containing large amounts of bona fide osteoclasts, showed large amounts of cross-reactive material, which diluted in parallel with the partially purified hairy cell leukemic TrACP in the ELISA. Commercial serum band 5b TrACP also displaced in parallel with the partially purified hairy cell leukemic TrACP. Immunoblotting studies revealed that the antiserum, but not nonimmune guinea pig serum, reacted with the homogeneous hairy cell leukemia splenic band 5 TrACPs, which were recently purified by our laboratory. Preliminary application of the ELISA to sera of patients with metabolic bone diseases revealed that normal healthy individuals had measurable amounts of the immunoreactive material, and patients with Paget's disease or hyperparathyroidism, who should have high bone turnover, had elevated levels of this immunoreactive material in their sera. In contrast, the level of serum osteoclastic TrACP in a patient with an acute lymphatic leukemia was normal. In summary, 1) we have shown that hairy cell leukemia splenic TrACP shares significant immunological similarity with the osteoclastic TrACP and with the serum band 5b TrACP, and 2) the ELISA holds promise for a sensitive and specific assay for bone resorption.

Topics: Acid Phosphatase; Animals; Bone Neoplasms; Chromatography, Affinity; Chromatography, Ion Exchange; Cross Reactions; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Histocytochemistry; Humans; Isoenzymes; Leukemia, Hairy Cell; Osteoblasts; Osteoclasts; Osteosarcoma; Rats; Spleen; Tartrates; Tumor Cells, Cultured

Tartrate resistant acid phosphatase (TRAP) is a reliable histochemical marker of osteoclasts when used on tissue sections of undecalcified bone. This paper presents an original morphometric analysis which can be done after histochemical identification of osteoclasts. These bone resorbing cells were demonstrated on undecalcified bone biopsies from control subjects and patients presenting a malignant disease of the lymphocyte B lineage. Computerized analysis of the osteoclastic population revealed that: (1) all TRAP positive cells along bone trabeculae belong to a osteoclastic population; (2) that B cell malignancies had an increased bone resorption. At the scanning electron microscopic level small resorption bays (about 10 microns in diameter) were observed either associated or separated from eroded surfaces presenting abnormal appearance; TRAP staining of histological sections of undecalcified bone, coupled with morphometric studies, may help in the understanding of bone disease pathobiology.

Topics: Acid Phosphatase; Bone and Bones; Bone Resorption; Humans; Leukemia, B-Cell; Leukemia, Hairy Cell; Leukemia, Lymphoid; Microscopy, Electron, Scanning; Osteoclasts

The author analyzes results of splenectomy in 20 patients with hairy cell leukosis. The importance of cytochemical investigations for the differential diagnosis is emphasized. Hematological indices normalized within two weeks after removal of the spleen. It is recommended to perform the operation of splenectomy in all patients with hairy cell leukosis.

Topics: Acid Phosphatase; Adult; Clinical Enzyme Tests; Female; Histocytochemistry; Humans; Leukemia, Hairy Cell; Male; Middle Aged; Preoperative Care; Spleen; Splenectomy

Tartrate-resistant acid phosphatases types 5a and 5b were purified from human hairy cell leukemia spleen by sequential chromatography on Phenyl-Sepharose, CM-Sepharose, concanavalin A-Sepharose, FPLC Superose-12 and FPLC Mono-S. The purification over the original tissue extract was 1150- and 3300-fold, with a yield of 2.1% and 2.5%, respectively. Gel filtration indicated an Mr of about 30000 for both forms. There was a N-terminal sequence identity between the two enzymes. However, they appeared to be different as assessed by cation exchange chromatography and amino acid composition.

Topics: Acid Phosphatase; Amino Acid Sequence; Amino Acids; Chromatography; Chromatography, High Pressure Liquid; Drug Resistance; Humans; Leukemia, Hairy Cell; Male; Middle Aged; Molecular Sequence Data; Molecular Weight; Spleen; Tartrates

The concomitant presence of B antigens and of the antigen recognized by the monoclonal antibody Leu-M5 (CD11c) on neoplastic lymphoid cells has been reported to be largely restricted to hairy cell leukemia (HCL). The authors studied Leu-M5 reactivity of neoplastic cells from 59 patients whose specimens were referred with a stated diagnosis of HCL by using the alkaline phosphatase anti-alkaline phosphatase technique on peripheral blood (PB) and bone marrow (BM) specimens. Tartrate-resistant acid phosphatase (AcP-T) activity was also studied. In 49 patients, HCL had been confirmed previously by BM biopsy, and specimens were evaluated for disease status during or after therapy with interferon (IFN) or 2'-deoxycoformycin. The remaining ten patients were newly referred for confirmation of the diagnosis of HCL before therapy. In all 55 patients in whom the BM biopsy demonstrated HCL, virtually every leukemic cell was Leu-M5 reactive, and the reaction proved, in some cases, to be helpful in the detection of small numbers of hairy cells in PB or BM preparations. AcP-T reactivity was demonstrated in the neoplastic cells of 52 of these 55 patients, including all but 3 of those receiving IFN, and was helpful in confirming persistent leukemia when interpretation of BM biopsy sections was difficult because the numbers of hairy cells were small. However, in four of the ten newly referred patients, BM biopsy showed features of splenic lymphoma with villous lymphocytes, rather than HCL. The neoplastic cells of these four patients were of B-cell origin and in three were Leu-M5 reactive. The authors' study indicates that Leu-M5 is present in nearly all hairy cells, but its presence in conjunction with other B-cell markers is not specific for HCL.

Topics: Acid Phosphatase; Alkaline Phosphatase; Antigens, Differentiation, T-Lymphocyte; Biopsy; Bone Marrow; Diagnosis, Differential; Humans; Immunoenzyme Techniques; Interferons; Leukemia, Hairy Cell; Lymphoproliferative Disorders

Topics: Acid Phosphatase; Bone Marrow; Humans; Leukemia, Hairy Cell; Male; Middle Aged; Primary Myelofibrosis

The tartrate-sensitive prostatic acid phosphatase, bands 2 and 4, are found in the soluble cytosol, and absent in the polysome of the prostate, while the tartrate-resistant acid phosphatase band 5 is present in the polysome and the soluble cytosol of hairy cells. The mRNA isolated from the prostate catalyzes the incorporation of 3T leucine into a protein different from that of bands 2 and 4. On the other hand, the mRNA isolated from the hairy cells catalyzes the incorporation of 3T leucine into band 5. The different biosynthetic mechanism of these two types of acid phosphatases are discussed in light of their different clinical significance.

Topics: Acid Phosphatase; Cell-Free System; Chromatography, High Pressure Liquid; Humans; Leukemia, Hairy Cell; Male; Polyribosomes; Prostate; RNA; RNA, Messenger; Spleen; Tartrates

B-cell neoplasias such as CLL can be viewed as models of monoclonal populations restricted within discrete ranges of B-cell maturation. It is unknown whether other B-cell leukemias such as prolymphocytic leukemia (PLL), lymphoplasmacytoid immunocytoma (IC), and hairy cell leukemia (HCL) involve different B lineages or are malignant variants of B cells in successive stages of development along the same lineage. Therefore in vitro maturation was induced with the phorbol ester TPA in leukemic cell samples from 10 CLL, 4 PLL, and 4 IC patients. Morphologically, both plasmacytic and hairy cell-like phenomena were induced. The latter unexpected finding was confirmed by reaction with HD6 (CD22) antibody which stains HCL but is unreactive with plasma cells, multiple myeloma, and CLL cells. Tartrate-resistant acid phosphatase was demonstrated in TPA-cultured CLL, PLL, and IC cells, and the same isoenzyme band as in HCL was revealed by isoelectrofocusing. On the other hand, an increase of IgM messenger RNA was detected in up to 20% of the cells in CLL cultures by single-cell in situ hybridization with fluoro-chrome-labeled DNA probes. An abundance of IgM messenger RNA characterizes lymphoplasmacytoid cells as found in IC. Our data demonstrate that CLL, PLL, and IC can be induced to realize a common genetic program which bears characteristics of HCL indicating that these four entities are more closely related than previously thought.

Topics: Acid Phosphatase; Antigens, Neoplasm; Humans; Immunoglobulin M; Isoenzymes; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphoma, Non-Hodgkin; Phenotype; RNA, Messenger; Tartrates; Tetradecanoylphorbol Acetate

Hairy cell leukemia (HCL) mononuclear cells were incubated with the phorbol ester TPA in an attempt to induce further maturation and were compared with B cell chronic lymphocytic leukemia, prolymphocytic leukemia, and non-Hodgkin's lymphoma cells. Morphology, surface features, membrane markers, tartrate-resistant acid phosphatase, and Ig secretion were examined. HCL cells spread and adhered firmly after TPA, producing elongated filopodia. Cells still retained ribosomal lamellar complexes, and increased numbers of dense bodies were seen. TPA enhanced the adherent and phagocytic properties of HCL cells, producing a modest increase in the expression of membrane Ig, GP-70, and Leu-M5 markers, tartrate-resistant acid phosphatase, and Ig secretion. Other neoplastic B cells behaved differently, forming readily detachable clumps without elongated filopodia. Maturation to plasma cells and hairy cell features were readily evident in all cases. These differences in growth patterns were consistent and may be used to distinguish HCL from other B cell neoplasias.

Topics: Acid Phosphatase; Antibodies, Monoclonal; Antigens, Differentiation; Antigens, Surface; Cell Membrane; Dose-Response Relationship, Drug; Humans; Immunoglobulins; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphoma, Non-Hodgkin; Microscopy, Electron, Scanning; Receptors, Antigen, B-Cell; Tartrates; Tetradecanoylphorbol Acetate

Mononuclear cells concentrated from 11 patients with chronic lymphocytic leukemia (CLL), 7 with non-Hodgkin's lymphoma in leukemic phase (NHL), 5 with hairy cell leukemia (HCL), 1 with prolymphocytic leukemia (PLL), and 1 with plasma cell leukemia (PCL) were induced to differentiate with various doses of TPA. The degree of induction was followed for up to 6 days by measuring the expression of surface membrane markers (SmIg and GP-70) and Ig secretion, the induction of tartrate-resistant acid phosphatase (TRAP) and by recording ultrastructural changes as seen by electronmicroscopy. The results show a dose and time dependency of the TPA effect and a great heterogeneity in the cellular response, particularly in cells obtained from B-CLL patients. TPA induced two main features, namely the development of "plasmacytoid" or "hairy cell" leukemia features that clearly depended on the dose and duration of treatment with the phorbol ester. The plasmacytoid features were more frequently encountered with lower doses (1 ng/ml) of TPA and were more evident after shorter exposures to TPA (1-2 days). Nevertheless, the hairy cell features were more striking after incubation with higher concentrations of TPA (10-100 ng/ml) after longer periods of incubation (up to 6 days) with lower doses of TPA. The various features of differentiation measured including cell morphology, surface membrane markers, Ig secretion, and TRAP staining, were frequently independent of each other, suggesting an autonomous pathway of differentiation for some of these features. Furthermore, in most of the cases, hairy cell leukemia features were obtained more frequently following TPA exposure than plasmacytic changes.

Topics: Acid Phosphatase; B-Lymphocytes; Cell Differentiation; Glycoproteins; Humans; Immunoglobulins; Isoenzymes; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphoma, Non-Hodgkin; Microscopy, Electron; Tetradecanoylphorbol Acetate

Cells from 11 chronic lymphocytic leukemic (CLL) patients were induced to differentiate with various doses of tetradecanoyl phorbol-13-acetate (TPA) and the degree of induction was followed up to six days by measuring the expression of two surface membrane markers (SmIg and GP-70), Ig secretion, tartrate-resistant acid phosphatase (TRAP), and ultrastructural changes. The results indicate dose and time dependency of the TPA effect and a great heterogeneity in the response to TPA among cells from different CLL patients. Furthermore, the two main TPA-induced features, the "plasmacytoid" or "hairy cell" features depended on the dose and duration of treatment with the phorbolester. The plasmacytoid features were more frequently encountered at low doses (1 ng/ml) of TPA and were evident after short exposures to TPA (1-2 days). The hairy cell features were more obvious after incubation with higher doses of TPA (10-100 ng/ml) or at Day 6 with lower doses of TPA. The differentiation features measured, including cell morphology, surface membrane markers, Ig secretion, and TRAP staining, appeared to be independent of each other suggesting an autonomous pathway of differentiation for some of these features.

Topics: Acid Phosphatase; Antibodies, Neoplasm; B-Lymphocytes; Dose-Response Relationship, Drug; Enzyme Induction; Humans; Leukemia, Hairy Cell; Leukemia, Lymphoid; Membrane Glycoproteins; Microscopy, Electron; Tetradecanoylphorbol Acetate; Time Factors

Tartrate-resistant acid phosphatase (T-AcP) has been used in the past 15 years as a specific test for the diagnosis of hairy cell leukemia (HCL). However, enzyme activity has been reported to be absent from the hairy cells of rare cases of HCL and to be present in the neoplastic cells of diseases other than HCL. In order to fully utilize T-AcP for the diagnosis of HCL, it is necessary to maximize the sensitivity and specificity of the staining method, to have adequate quality control to ensure technical and interpretative accuracy, and to prepare optimal cytologic and histologic materials for study. In blood, only the presence of cells with intense T-AcP activity is diagnostic of HCL (positive T-AcP test). In tissues other than blood, the presence of cells with intense enzyme activity may not be diagnostic for HCL; assessment of cell morphology and appreciation of the pattern of enzyme localization are also important. In studying the blood of over 1,000 patients, we have found a negative T-AcP test in two of 200 cases of HCL and a positive T-AcP test in three of 800 patients with diseases other than HCL. We believe that the T-AcP test, when performed and interpreted properly, is a useful diagnostic test for HCL.

Topics: Acid Phosphatase; Bone Marrow; Histocytochemistry; Humans; Leukemia, Hairy Cell; Leukocytes; Liver; Lymph Nodes; Spleen; Tartrates

Hairy cell leukaemia is a form of leukaemia difficult to diagnose since pancytopenia is often present. Hairy cells contain tartrate-resistant acid phosphatase, and this factor is utilised in the diagnosis of the condition. This study confirms that it is also possible to demonstrate tartrate-resistant acid phosphatase in leukaemic infiltrates in formalin fixed paraffin-embedded tissue sections.

Topics: Acid Phosphatase; Clinical Enzyme Tests; Histocytochemistry; Humans; Leukemia, Hairy Cell; Liver; Lymph Nodes; Paraffin; Pronase; Spleen; Staining and Labeling; Tartrates

In vitro maturation was induced with 12-O-tetradecanoylphorbol-13-acetate in leukemic cell samples from patients with chronic lymphocytic leukemia (n = 10) and prolymphocytic leukemia (n = 4). The cells were studied for morphology, for immunological markers using the fluorescence activated cell sorter, and for acid phosphatase isoenzymes using both cytochemistry and isoelectrofocusing. Morphologically the induced changes included appearance of cells with an excentric nucleus and basophilic cytoplasm and eventually of cells with many fine cytoplasmic projections ("hairs"). Analysis of immunological markers by flow cytometry revealed that the monoclonal antibody defined cell surface molecule HD6 (CD22), which is strongly expressed on hairy cell leukemia (HCL) but absent from plasmacytoma and plasma cells, can be induced or enhanced in the leukemic samples. In the study of acid phosphatase isoenzymes using cytochemistry we observed the induction of the tartrate resistant isoenzyme. Further, using isoelectrofocusing we could demonstrate the induction of the same band of tartrate resistant acid phosphatase with an isoelectric point of 9.0-9.7 as detected also in HCL. This particular isoenzyme is considered characteristic of HCL but is absent in plasmacytoma. Our data demonstrate that chronic lymphocytic leukemia and prolymphocytic leukemia cells can be induced to realize a common genetic program which bears characteristics of HCL, indicating that these three entities are much more closely related than previously thought.

Topics: Acid Phosphatase; Aged; Cells, Cultured; Female; Humans; Leukemia, Hairy Cell; Leukemia, Lymphoid; Male; Middle Aged; Phenotype; Receptors, Antigen, B-Cell; Tetradecanoylphorbol Acetate

The differential diagnostic significance of acid phosphatase and beta-glucuronidase were studied in 77 cases of low-grade B cell non-Hodgkin's lymphomas. In most cases the results of cytochemical enzyme studies performed on malignant cells of the bone marrow were evaluated. B cell chronic lymphocytic leukaemia, centrocytic and centroblastic/centrocytic lymphomas were characterized by a weak or a negative acid phosphatase and beta-glucuronidase activity. Stronger positivity was observed in immunocytoma and in Waldenström's macroglobulinaemia, while the highest activity was found in multiple myeloma. Hairy cell leukaemia of B cell origin showed intensive tartrate-resistant acid phosphatase activity. The cytochemical examination of these lysosomal enzymes may be useful in the diagnosis of low-grade malignant lymphomas of B cell origin by completing other methods.

Topics: Acid Phosphatase; Adult; B-Lymphocytes; Bone Marrow; Glucuronidase; Humans; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphoma; Lymphoma, Follicular; Lysosomes; Multiple Myeloma; Waldenstrom Macroglobulinemia

We have investigated the relationship between chronic lymphocytic leukemia (CLL), hairy cell leukemia (HCL), and different normal B cell subsets: Mrbc+, T1+ and slgM+ tonsil cells; germinal center; mantle zone; and peripheral blood B lymphocytes. Both malignant and normal cells were incubated in vitro with the phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA) for 72 hours and the morphology, cytochemical profile, and surface markers were evaluated. The results show that CLL cells TPA-induced become indistinguishable from HCL by four independent criteria: the morphology; the cytoplasmic tartrate resistant acid phosphatase (TRAP) enzyme activity; the membrane positivity with anti-Leu M5 (SHCL3); and anti-Tac monoclonal antibodies which, in the uninduced state, react only with HCL. The features of TRAP and Tac positivity are also expressed (though in variable degree) by different normal B cell populations activated with TPA or pokeweed mitogen (PWM). It is concluded that HCL might represent an aberrantly activated variant of CLL (or of a CLL-related disorder).

Topics: Acid Phosphatase; Aged; B-Lymphocytes; Bone Marrow Cells; Cells, Cultured; Embryonal Carcinoma Stem Cells; Humans; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphocyte Activation; Male; Middle Aged; Mitogens; Neoplastic Stem Cells; Phorbol Esters

We examined the fine structure and enzymatic activity of cells composing colonies grown from blood and/or spleen of eight patients with hairy cell leukemia. Mononuclear cells (MNC) were plated over an agar layer in a medium containing methylcellulose and leukocyte-conditioned medium. After 7-10 days incubation colonies were harvested for cytologic study. Colony cells possessed a euchromatic nucleus with an occasional nucleolus. Their cytoplasm contained a prominent Golgi region, numerous mitochondria and small vesicles, many short strands of rough endoplasmic reticulum (RER) and an infrequent phagosome. Well-developed ribosome-lamella complexes and what may have been their intermediate forms appeared in colony cells from three patients. Strong activity for tartrate-resistant acid phosphatase, localized in the RER, nuclear envelope and some Golgi vesicles, was evident in 50-95% of all colony cells. Our results indicate that a high proportion of MNC forming colonies in this culture system maintain the characteristic morphology and cytochemical activity of hairy cells.

Topics: Acid Phosphatase; Cells, Cultured; Endoplasmic Reticulum; Histocytochemistry; Humans; Leukemia, Hairy Cell; Microscopy, Electron; Neoplastic Stem Cells

The spleens of patients with hairy cell leukemia contain high levels of a tartrate-insensitive, cationic, acid phosphatase (the human Type 5 isozyme). This phosphatase has been purified by a procedure which involves only two chromatographic steps: CM-cellulose chromatography and immunoaffinity chromatography on sheep antibodies generated against porcine uteroferrin. Uteroferrin is an abundant iron-containing acid phosphatase that can be recovered readily from porcine uterine secretions. Like uteroferrin, the purified human Type 5 phosphatase is a glycoprotein of molecular weight about 34,000. It contains two atoms of iron/molecule. The human phosphatase and uteroferrin also resemble each other closely in electrophoretic mobility, substrate specificity, and response to a variety of activators and inhibitors. Mouse monoclonal antibodies have been raised to uteroferrin and to the human Type 5 phosphatase. Three monoclonal antibodies which bind with high affinities to distinct sites on the uteroferrin molecule also recognize the human spleen enzyme, but bind to it with much lower affinity. These antibodies also recognize cationic acid phosphatases purified from bovine and rat spleens. A monoclonal antibody raised against the human enzyme, but selected for binding to uteroferrin, appears to recognize a relatively conserved site on all four phosphatases. We conclude that the human Type 5 isozyme belongs to a growing class of structurally related, iron-containing acid phosphatases which includes the iron-transport protein, uteroferrin.

Topics: Acid Phosphatase; Animals; Antibodies, Monoclonal; Binding, Competitive; Cattle; Chromatography, Ion Exchange; Electrophoresis, Polyacrylamide Gel; Humans; Isoenzymes; Leukemia, Hairy Cell; Male; Mercaptoethanol; Metalloproteins; Middle Aged; Rats; Spleen; Substrate Specificity; Swine; Tartrate-Resistant Acid Phosphatase

We examined cellular relationships and cytokinetics of hairy cells in colonies formed in an in vitro cloning system. Cells in small colonies and at the periphery of larger ones were separated by wide, irregular intercellular spaces. Cells in the core of large colonies were elaborately intertwined by their cytoplasmic processes and so densely packed that their intercellularity was greatly reduced. Cell labeling with 3H-thymidine revealed indices ranging from less than 1% to 35%. The combination of autoradiography with a stain for tartrate-resistant acid phosphatase showed that the enzyme was not expressed by cells in S-phase. The cellular relationships of hairy cells in colonies grown in this system closely mimic those of their in vivo counterparts in the spleen and bone marrow.

Topics: Acid Phosphatase; Autoradiography; Cell Communication; Cell Division; Cell Separation; Clone Cells; Culture Media; Humans; Kinetics; Leukemia, Hairy Cell; Microscopy, Electron; Microvilli; Pseudopodia; Tartrates

Bone marrow specimens from 21 patients with hairy cell leukemia (HCL) who were entered into a program to study the efficacy of treatment with recombinant alpha 2-interferon were evaluated. Patients were treated with the interferon, 2 X 10(6) U/m2 subcutaneously three times weekly, and were scheduled to undergo bone marrow aspiration and biopsy at study entry and after three (21 patients) and six (16 patients) months of treatment. Bone marrow samples after three months of treatment showed an overall decline in cellularity, from an average of 77 +/- 20 to 57 +/- 22 per cent, with a marked decrease in the percentage of neoplastic mass (from 87 +/- 9 to 59 +/- 24 per cent). The bone marrow changes were associated with significant improvement in hematologic values, including hemoglobin levels and granulocyte and platelet counts. The bone marrow changes and improved hematologic values remained stable with continuation of interferon therapy. However complete bone marrow remission did not occur in any of the patients after three or six months of interferon therapy. The HCL cell mass in more than 60 per cent of the patients remained at or above 50 per cent of the marrow cellularity and dropped to less than 25 per cent in 14 per cent of the patients. In all of the patients increased amounts of reticulin fibers were identified in the bone marrow prior to therapy, and 89 per cent of bone marrow aspirations failed (dry tap). The amounts of reticulin fibers remained increased in most of the patients (91 per cent), with a high incidence of dry taps (73 per cent), after therapy. Interferon therapy also changed the tartrate-resistant acid phosphatase(TRAP)-positive HCL cells to TRAP-negative, suggesting inhibition of activity and/or production of TRAP in HCL cells.

Topics: Acid Phosphatase; Adult; Aged; Bone Marrow; Cell Count; Female; Hematopoiesis; Humans; Interferon Type I; Leukemia, Hairy Cell; Male; Middle Aged; Recombinant Proteins; Reticulin; Tartrates; Time Factors

Topics: Acid Phosphatase; Antigens, Surface; Biopsy; Bone Marrow; Diagnosis, Differential; Female; Humans; Immunoglobulins; Isoenzymes; Leukemia, Hairy Cell; Leukocyte Count; Male; Middle Aged; Receptors, Fc

The subcellular localization of acid phosphatase (AcP) in various immunologically-defined neoplastic lymphoid cells including hairy cells was investigated by electron microscopy. 2 substrates, naphthol-AS-BI-phosphoric acid (naphthol-AS-BI-P) and sodium beta-glycerophosphate, were compared. By incubation in naphthol-AS-BI-P containing medium, the reaction product was found located in granules, vesicles, the Golgi apparatus, the rough ER including the nuclear envelope in the cells of T ALL, T CLL and HCL. A typical pattern of reaction was observed for each of these disorders: enzyme-positive Golgi membranes and neighbouring granules, clustered in the nuclear notch in T cell-derived lymphoblasts; enzyme-positive granules around Gall bodies, aggregated paranuclearly in T CLL lymphocytes and enzyme-positive scattered cytoplasmic granules and vesicles in hairy cells. Enzyme activity was occasionally seen in singly-occurring granules in the cells of cALL, B CLL and B PLL, rarely in other substructures. With the Gomori method using beta-glycerophosphate as substrate, the enzyme reaction was limited primarily to lysosomal sites and was seldom observed in other organelles. Tartrate-resistant AcP was found in the majority of hairy cells and in a few prolymphocytes, located in the same structures as AcP without tartrate.

Topics: Acid Phosphatase; B-Lymphocytes; Cytoplasmic Granules; Glycerophosphates; Golgi Apparatus; Humans; Leukemia, Hairy Cell; Leukemia, Lymphoid; Microscopy, Electron; Organophosphorus Compounds; T-Lymphocytes

Topics: Acid Phosphatase; Biopsy; Humans; Leukemia, Hairy Cell; Liver; Tartrates

The leukemic infiltrate in the liver in hairy cell leukemia may be easily overlooked in conventional histological slides and an exact classification may not always be possible. In order to facilitate the identification of tricholeukocytes , it is suggested to perform the tartrate-resistant acid phosphatase stain on routinely formalin fixed and paraffin-embedded liver biopsy specimens. Thereby it will be in most cases possible to demonstrate sufficiently enough enzyme activity to establish unequivocally the diagnosis of hairy cell leukemia.

Topics: Acid Phosphatase; Biopsy, Needle; Histocytochemistry; Humans; Leukemia, Hairy Cell; Leukocytes; Liver

Six patients with hairy cell leukemia (HCL) were studied for surface immunoglobulin ( sIg ). In all five sIg -positive cases, the heavy chain isotype was IgG. We performed cytologic and cytochemical studies of sIgG + lymphocytes in normal peripheral blood and compared them with hairy cells. Normal sIgM + lymphocytes were also examined. sIgG + and sIgM + lymphocytes made up 0.9% and 6.1% of normal peripheral blood lymphocytes, respectively. Under a phase-contrast microscope, 76% of sIgG + lymphocytes showed cytoplasmic processes similar to those found on hairy cells, whereas most sIgM + lymphocytes had smooth surfaces. Tartrate-resistant acid phosphatase (TRAP) staining revealed that TRAP-positive cells accounted for 65% of sIgG + lymphocytes and 19% of sIgM + lymphocytes. Some (8.3%) of the sIgM + lymphocytes expressed sIgG concomitantly. When sIgM +, sIgM +, sIgG + lymphocytes were excluded, the percentages of cells with surface processes and of TRAP-positive cells in the remaining sIgM +, sIgG - lymphocytes were 10% and 12%, respectively. A very small proportion (0.2%) of sIgM -, sIgG - lymphocytes had cytoplasmic processes. These results indicate that normal sIgG + lymphocytes are cytologically and cytochemically different from most sIgM + lymphocytes and that the phase-contrast microscopic appearances and TRAP activity of sIgG + lymphocytes are similar to those of HCL tumor cells.

Topics: Acid Phosphatase; Cell Membrane; Humans; Immunoglobulin G; Immunoglobulin M; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphocytes; Male; Receptors, Antigen, B-Cell

In order to assess the role of Epstein-Barr virus (EBV) in patients with hairy cell leukemia (HCL), we have sought to characterize 1) the ability of EBV to infect and transform hairy leukemic cells in vitro and 2) the phenotypes of cell lines putatively derived from those leukemic cells. Analysis of EBV-induced transformation and the kinetics of Epstein-Barr nuclear antigen (EBNA) induction in leukemic preparations indicated that most leukemic cells were not susceptible to EBV infection but that at least a small subpopulation of leukemic cells could be infected with EBV. Lymphoblastoid cells lines were established after exposure of peripheral blood or splenic cells from HCL patients to B95-8 or QIMR-WIL EBV. Splenic leukemic cell preparations were more sensitive targets for EBV transformation than were peripheral blood cell samples. The newly established cell lines, but not long-established B lines such as Raji, demonstrated high levels of synthesis of p35, (a protein complex expressed abundantly by cells of a subset of HCL patients) and high levels of tartrate-resistant acid phosphatase (an enzyme relatively diagnostic for HCL). Lymphoblastoid lines from one patient with HCL expressed lambda light chains and no kappa chains as did the patient's leukemic cells. Virus expression in these lines showed that HCL-derived lines had spontaneous early antigen (EA) and viral capsid antigen (VCA) expression. Transforming EBV could be rescued from HCL-derived cell lines but not from cord blood-derived lines.

Topics: Acid Phosphatase; Antigens, Neoplasm; Antigens, Surface; Antigens, Viral; Capsid; Cell Line; Cell Transformation, Viral; Electrophoresis, Polyacrylamide Gel; Herpesvirus 4, Human; Humans; Immunoglobulins; Leukemia, Hairy Cell; Neoplasm Proteins; Phenotype

Topics: Acid Phosphatase; Glucuronidase; Humans; Leukemia, Hairy Cell

Nineteen patients with hairy cell leukemia were studied in order to define the hepatic changes in this disease and to correlate the morphologic changes in the liver with the clinical and biochemical findings. Although only eight of the patients had hepatomegaly, all 19 had microscopic mononuclear cell infiltration in the sinusoids or the portal areas or both. The severity of mononuclear infiltration in the liver correlated poorly with the size of the liver or spleen, the biochemical changes, or number of hairy cells in the blood. Abnormal serum biochemical values were present occasionally and were usually due to associated diseases or to complications of this disease. Elevated serum alkaline phosphatase activity was noted in four patients; three of them had granulomatous lesions in the liver. Unless the characteristic "clear cell" pattern is seen, the hepatic mononuclear cell infiltration may not be diagnostic of hairy cell leukemia and, in many instances, may not even be suggestive of neoplasia. A new technique for demonstrating tartrate-resistant acid phosphatase activity in methacrylate-embedded sections was developed, which allowed identification of hairy cells in the liver biopsy specimens of all five patients so studied. The authors concluded that liver involvement is common in this disease.

Topics: Acid Phosphatase; Adult; Aged; Alkaline Phosphatase; Biopsy; Female; Histocytochemistry; Humans; Leukemia, Hairy Cell; Leukocyte Count; Liver; Male; Middle Aged; Organ Size

A patient is described in whom cutaneous lesions were the initial manifestation of hairy-cell leukemia. Touch preparations made immediately on removal of a 3-mm punch biopsy specimen of the cutaneous lesions revealed acid-phosphatase positive, tartrate-resistant staining in the leukemic cells, and helped to establish the diagnosis. Specific eruptions occur in approximately 7% of patients with hairy-cell leukemia, appearing grossly as disseminated, erythematous maculopapules, with a perivascular mononuclear leukemic cell infiltrate seen microscopically. A review of the English literature indicates that cutaneous manifestations are not generally recognized as a diagnostic source in individuals with hairy-cell leukemia, and biopsy is seldom undertaken.

Topics: Acid Phosphatase; Humans; Leukemia, Hairy Cell; Male; Middle Aged; Skin; Tartrates

This article reviews the authors' experience with hairy cell leukemia to determine the incidence of hypoplastic marrows in this disease and the utiity of embedding marrow biopsy specimens in plastic, which allows the sections to be stained for tartrate-resistant acid phosphatase (TRAP). It was found that the presence of hairy cells in the peripheral blood, marrow aspirate, or both established the diagnosis in eight of 11 patients; in the remaining three patients, the marrow biopsy specimen was necessary for definitive diagnosis. The bone marrow was markedly hypocellular in three patients, including one in whom the marrow morphologic results were essential for diagnosis. Embedding the marrow biopsy specimen in plastic and staining the sections for TRAP was noted to be a useful technique in establishing the diagnosis of hairy cell leukemia. In all cases in which this was performed, the authors found abundant TRAP-positive cells in the marrow biopsy specimen; even in those cases with markedly hypocellular marrows, this technique revealed the presence of hairy cells. In one patient, TRAP staining of the marrow biopsy specimen proved essential for definitive diagnosis. The authors concluded that the occurrence of marrow hypocellularity is not rare in patients with hairy cell leukemia and that TRAP-staining of plastic-embedded marrow biopsy specimens is a helpful and sometimes essential technique in diagnosing hairy cell leukemia.

Topics: Acid Phosphatase; Aged; Biopsy; Bone Marrow; Clinical Enzyme Tests; Diagnosis, Differential; Histological Techniques; Humans; Leukemia, Hairy Cell; Male; Staining and Labeling

In a case of hairy cell leukemia the evidence of tartrate-resistant isoenzyme 5 of phosphatase (TRISP 5) developed to findings important for the diagnosis. The evidence was achieved by means of disc electrophoresis by separating the cell lysate of white blood cells. The percentage of TRISP 5 in the total activity of acid phosphatase in leukocytes amounted to 17.6%. In cytochemical respect the evidence of TRISP 5 proved to be negative. The classification of these findings and their differential-diagnostic significance are discussed.

Topics: Acid Phosphatase; Diagnosis, Differential; Electrophoresis, Disc; Humans; Isoenzymes; Leukemia, Hairy Cell; Leukocytes; Middle Aged; Tartrates

Acid phosphatase (AcP) in neoplastic cells from various lymphoid leukemias was examined. In the cytochemical studies, tartrate-resistant AcP (T-rAcP) activity was observed in the neoplastic cells from well-differentiated lymphoid leukemias such as adult T-cell leukemia (ATL), B-cell chronic lymphocytic leukemia (B-CLL), T-cell chronic lymphocytic leukemia (T-CLL), and hairy-cell leukemia (HCL). T-rAcP activity was also detected in a small number of leukemic cells obtained from T-cell acute lymphoblastic leukemia (T-ALL), while it was not detected in the neoplastic cells from null-ALL, macroglobulinemia, and multiple myeloma (MM). In the electrophoretical studies, fraction 1 (F-1), F-3, F-3b, and F-4 were completely tartrate-sensitive, while F-2 was partially resistant and F-5 was completely resistant. T-rAcP activity (F-5) was observed in ATL cells, B-CLL cells, and HCL cells, while it was not detected in ALL cells, macroglobulinemia cells, and MM cells. The present study indicates that T-rAcP activity is observed not only in HCL cells but also in the well-differentiated lymphoid cells such as ATL cells, B-CLL and T-CLL cells except the most highly differentiated forms of B-cells of MM and macroglobulinemia.

Topics: Acid Phosphatase; Cell Transformation, Neoplastic; Electrophoresis, Polyacrylamide Gel; Humans; Isoenzymes; Leukemia, Hairy Cell; Leukemia, Lymphoid; Multiple Myeloma; T-Lymphocytes; Tartrates; Waldenstrom Macroglobulinemia

Four acid hydrolases, acid phosphatase (AP), alpha-naphthyl acetate esterase (ANAE), beta-glucuronidase, and N-acetyl-beta-glucosaminidase, were determined cytochemically in peripheral blood lymphocytes from 50 patients with B and T chronic lymphocytic and prolymphocytic leukemias (CLL, PLL) and related disorders. Strong positive reactions were characteristic of the T-cell leukemias while the reactions were weak or negative in B-CLL and B-PLL. Differences in the cytochemical profile of T-CLL and T-PLL were noted. In both, beta-glucuronidase and N-acetyl-beta-glucosaminidase were positive; these enzymes are therefore good cytochemical markers of the chronic T-cell leukemias. AP and ANAE gave different results according to the disease process; AP was strong in T-CLL and variable in T-PLL, while ANAE was strongly positive in T-PLL, but weak or negative in T-CLL. The findings in T-CLL, a proliferation of T gamma lymphocytes, were similar to those of normal T gamma cells. In T-PLL, the findings did not relate to the membrane phenotype as defined by monoclonal antibodies.

Topics: Acetylglucosaminidase; Acid Phosphatase; B-Lymphocytes; Glucuronidase; Histocytochemistry; Humans; Hydrolases; Leukemia, Hairy Cell; Leukemia, Lymphoid; Naphthol AS D Esterase; Rosette Formation; T-Lymphocytes

Tartrate-resistant acid phosphatase was isolated from serum and spleen of patients affected by Gaucher's disease. Electrophoretic and antigenic properties were compared to the enzyme isolated from hairy cells described in a previous study (9). The enzyme isolated from Gaucher serum has electrophoretic and antigenic properties identical to the acid phosphatase band 5b of hairy cells. The major tartrate-resistant acid phosphatase in the Gaucher spleen is band 5a. Bands 5a and 5b have identical protein structure indicated by their identical antigenicity. The removal of carbohydrate from band 5a by sialidase converted band 5a to 5b.

Topics: Acid Phosphatase; Adolescent; Adult; Blood Protein Electrophoresis; Child; Child, Preschool; Colorimetry; Female; Gaucher Disease; Humans; Immunodiffusion; Leukemia, Hairy Cell; Male; Middle Aged; Molecular Weight; Tartrates

Topics: Acid Phosphatase; Female; Humans; Leukapheresis; Leukemia, Hairy Cell; Lymphocyte Activation; Lymphocytes; Microscopy, Electron, Scanning; Middle Aged; Mitogens; Receptors, Antigen, B-Cell

A strong activity of the tartrate-resistant acid phosphatase, isoenzyme 5b, was observed in mouse spleen. The enzyme of the mouse spleen had similar electrophoretic mobility and related antigenicity to that of the reticulum cells of leukemic reticuloendotheliosis. Enzyme histochemical studies showed many non-phagocytic cells with strong tartrate-resistant acid phosphatase in spleen. The data suggest that mouse tissue is a good animal model to study the normal cell type with strong tartrate-resistant acid phosphatase.

Topics: Acid Phosphatase; Animals; Histocytochemistry; Humans; Leukemia, Hairy Cell; Mice; Neoplasms, Experimental; Reticulocytes; Spleen; Tartrates

Seven patients are presented with a chronic lymphoproliferative disorder characterized clinically by splenomegaly, no or discrete lymphnode enlargement, and a varying degree of cytopenia. In blood and bone-marrow smears lymphoid cells of "hairy" appearance are demonstrable which may contain tartrate-resistant acid phosphatase. The finding of a nodular bone-marrow infiltration without fibrosis as well as that of a nodular infiltration of the spleen originating in the white pulp are incompatible with the diagnosis hairy-cell leukemia and place the disease near to chronic lymphocytic leukemia (CLL) or leukemic immunocytoma respectively. A detailed cytologic and cytochemical examination of the infiltrating cells shows deviations from the typical enzymatic pattern of hairy cells and from known enzymatic constellations in CLL and related lymphoproliferative disorders. Thus, we are dealing with an intermediate form, difficult to classify, the separation of which nevertheless seems to be important for therapeutical reasons.

Topics: Acid Phosphatase; Aged; Bone Marrow; Chronic Disease; Diagnosis, Differential; Female; Humans; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphoproliferative Disorders; Male; Middle Aged; Spleen; Splenomegaly

A father and son hairy cell leukemia are identified on the basis of tartrate-resistant acid-phosphatase-positive hairy cells and diagnostic bone core biopsies. Personal interviews of the two patients revealed no evidence of consanguinity, primary immune deficiency, or common medication use. HLA typing revealed both patients to be A1,3 and B8,14.

Topics: Acid Phosphatase; Aged; Blood Cell Count; Bone Marrow; Histocompatibility Testing; Humans; Leukemia, Hairy Cell; Male; Middle Aged

We purified acid phosphatase isoenzyme 5b from a human spleen affected by leukemic reticuloendotheliosis and used it to produce a specific antiserum. The antiserum was used to show complete immunological identity among isoenzymes 5a and 5b in human serum, and 5b isolated from a giant-cell bone tumor and from the spleen of a case of Hodgkin's disease. Acid phosphatase 5b in a giant-cell bone tumor was isolated for biochemical characterization. Its pH optimum and substrate specificity were very similar to those of isoenzyme 5b from human spleen.

Topics: Acid Phosphatase; Bone and Bones; Hodgkin Disease; Humans; Hydrogen-Ion Concentration; Immunodiffusion; Isoenzymes; Kinetics; Leukemia, Hairy Cell; Spleen; Substrate Specificity; Tartrates

Hairy-cell leukemia is characterized clinically in splenomegaly and pancytopenia and pathologically by the proliferation in hematopoietic tissue of cells containing the tartrate-resistant isozyme 5 of acid phosphatase. We have described a patient with a T-lymphocyte variant of this disease. A permanent cell line obtained from the spleen of this patient has the biological and enzymatic characteristics of the fresh leukemic cells. We have used this line to study the surface morphology, ultrastructure, and ultrastructural localization of acid phosphatase in defined T-lymphoid hairy cells. The surface of the cells of the permanent line was smooth but many hair-like projections appeared after exposure to phytohemagglutinin (PHA). There was little acid phosphatase reaction produce visualized when beta-glycerophosphate was used as a substrate. With sodium haphthol AS-BI phosphoric acid heavy deposits were seen in the perinuclear membrane, mitochondria, and rough endoplasmic reticulum. Exposure to PHA and pokeweed mitogen resulted in increased reaction product, suggesting increased enzyme synthesis. Tartrate-resistant acid phosphatase was localized in the same organelles.

Topics: Acid Phosphatase; Cell Line; Histocytochemistry; Humans; Leukemia, Hairy Cell; Microscopy, Electron, Scanning; T-Lymphocytes; Tartrates

Topics: Acid Phosphatase; Histological Techniques; Humans; Leukemia; Leukemia, Hairy Cell; Tartrates

Topics: Acid Phosphatase; Butyrates; Carboxylic Ester Hydrolases; Histocytochemistry; Humans; Leukemia; Leukemia, Hairy Cell; Periodic Acid-Schiff Reaction; Tartrates

Tartrate-resistant acid phosphatase activity may be demonstrated in paraffin-embedded liver and spleen specimens as well as in decalcified bone marrow biopsies after fixation in a mixed glutaraldehyde-formaldehyde-calcium acetate solution. This technique may routinely be applied to haematologically relevant material and aids in the differential diagnosis of hairy-cell leukaemia.

Topics: Acid Phosphatase; Bone Marrow; Diagnosis, Differential; Histocytochemistry; Humans; Leukemia, Hairy Cell; Liver; Spleen; Tartrates

The Kiel classification of lymphomas based on cytological criteria can be applied to tumor cell types found in liver infiltrates as well. Lymphomas with low degree of malignancy are delineated distinctly against the normal parenchyma; the borderline is slightly blurred only in the centroblastic-centrocytic subtype. Generally, the portal fields in these cases are round-shaped, expanded, and filled with lymphoid cells. Lymphomas with a high degree of malignancy are characterized on the contrary by portal fields with very irregular borderlines; in some cases there is a certain similarity to piece meal necrosis. There is an increased incidence of mitoses in the malignant tissue. Bile ducts are often disrupted. Hair cell leukemia can be differentiated in paraffin sections too, if tartrate-resistent acid phosphotase is present.

Topics: Acid Phosphatase; Bile Ducts, Intrahepatic; Diagnosis, Differential; Humans; Leukemia, Hairy Cell; Liver Neoplasms; Lymphoma; Mitosis; Neoplasm Invasiveness

Topics: Acid Phosphatase; Cell Survival; Humans; Hydrogen-Ion Concentration; Isoelectric Focusing; Leukemia, Hairy Cell; Lymphocytes; Lysosomes; Spleen

Over a 4-year period 203 patients with various types of leukaemia were treated by the Haematology Unit at the Johannesburg Hospital. Ten of them were suffering from the condition known as hairy-cell leukaemia or leukaemic reticulo-endotheliosis. They were all men, and ranged in age from 29 to 67 years (mean 56 years). The majority presented with pancytopenia, and there was invariably splenomegaly, while lymphadenopathy was rare. Hairy cells were identified microscopically in the peripheral blood of 7 patients and in 5 the specific cytochemical marker, tartrate-resistant acid phosphatase, was present. In addition, in a further 2 patients this feature, which was not identified in the peripheral blood, was found in the splenic cells. The bone marrow trephine biopsy specimens characteristically showed extensive lymphoid infiltration associated with a dense disordered deposition of reticulin fibres. Electron microscopical and immunological studies proved to be of doubtful diagnostic value. Splenectomy was carried out on 9 patients, and there was tumour involvement in all the spleens removed. Two patients died from septicaemia, the one before splenectomy and the other 9 months after the operation. The 8 remaining patients have had their subjective symptoms alleviated and their peripheral blood indices have been improved by splenectomy, and none has required further treatment for periods now ranging from 7 to 41 months.

Topics: Acid Phosphatase; Adult; Aged; Blood Cell Count; Bone Marrow Cells; Hemoglobins; Humans; Leukemia, Hairy Cell; Lymphocytes; Male; Middle Aged; Organ Size; Rosette Formation; Spleen; Splenectomy; Staining and Labeling

Cytochemical and immunological studies were performed on tissues and mononuclear cell suspensions from ten patients with hairy cell leukemia. In all cases studied, tartrate-resistant acid phosphatase was noted within the cytoplasm of hairy cells (HCs). In two thirds of the cases, alpha naphthyl acetate esterase was observed in HCs. In addition, HCs did not form spontaneous rosettes with sheep erythrocytes. A variable number of HCs displayed complement receptors. The nonspecific binding of conjugated immunoglobulin to HCs probably reflected the presence of a high concentration of Fc receptors on the HCs surface. In three cases, the conjugated immunoglobulin reacted predominantly to one light chain, thus suggesting the presence of a monoclonal immunoglobulin. In four of six cases, mononuclear cell suspensions of HCs demonstrated latex phagocytosis. In one case, HCs displayed resynthesis of surface membrane immunoglobulin, the presence of B cell antigen, and phagocytosis of latex. These findings suggest that HCs are distinctive and contain properties of both B lymphocytes and monocytes.

Topics: Acid Phosphatase; B-Lymphocytes; Humans; Leukemia, Hairy Cell; Phagocytosis; Receptors, Antigen, B-Cell; Rosette Formation

Isoenzymatic study of leucocytic acid phosphatase under normal conditions identifies 3 isoenzymatic bands, which exhibit a noticeable cell specificity. Band 2 is granulocytic, band 3 lymphocytic and band 4 monocytic in origin. Pathologic deviations in the isoenzymatic pattern are both qualitative and quantitative. For some diseases such as chronic lymphocytic leukaemia there is a well-defined, differential pattern according to immunological B- or T-cell origin. The more significant qualitative aspects are related to the appearance of abnormal bands, especially band 3b, indicating blastic cellularity, and 5, corresponding to hairy cells. The isoenzymatic analysis of acid phosphatase activity is a simple haematologic complementary test, particularly useful in the differential diagnosis of lymphoproliferative disorders with peripheral blood manifestations.

Topics: Acid Phosphatase; B-Lymphocytes; Diagnosis, Differential; Electrophoresis, Cellulose Acetate; Humans; Isoenzymes; Leukemia; Leukemia, Hairy Cell; Leukemia, Lymphoid; Leukocytes; Lymphocytes; Lymphoma, Non-Hodgkin; Sezary Syndrome; T-Lymphocytes

Topics: Acid Phosphatase; Fluorescent Antibody Technique; Humans; Leukemia, Hairy Cell; Microscopy, Phase-Contrast; Receptors, Antigen, B-Cell; Rosette Formation; Spleen

Topics: Acid Phosphatase; Histocytochemistry; Indicators and Reagents; Leukemia, Hairy Cell; Leukocytes; Tartrates

Among 12 patients with hairy cell leukaemia two died without splenectomy and two more 11 months and 7 1/2 years after the operation. In all patients removal of hypersplenism led to improvement of haematological criteria over several months up to many years. This was particularly true for platelets and granulocytes and thus for the proneness to infection and haemorrhagic diathesis. In two patients normalisation of all haematological parameters including histologically scarcely detectable hairy cells was observed. Differential diagnosis from prolymphocyte leukaemia is difficult. Nuclear morphology permits recognition of different degrees of maturation of hairy cells. In cases where tartrate resistant acid phosphatase (TRAP) cannot be demonstrated cytologically, demonstration of isoenzyme 5 is sometimes possible using gel electrophoresis. Demonstration of TRAP remains valuable for the diagnosis of hairy cell leukaemia, however only in connection with other criteria indicating this form of leukaemia.

Topics: Acid Phosphatase; Adult; Aged; Female; Humans; Hypersplenism; Isoenzymes; Leukemia, Hairy Cell; Male; Middle Aged; Splenectomy

Topics: Acid Phosphatase; Leukemia, Hairy Cell; Spleen; Tartrates

Topics: Acid Phosphatase; Alkaline Phosphatase; Humans; Leukemia, Hairy Cell; Neutrophils

Cytochemical demonstration of tartrate-resistant acid phosphatase activity is essential for the diagnosis of leukemic reticuloendotheliosis. In order to perform this test correctly and to interpret the results propertly, it is necessary to understand the technical details of the cytochemical methods thoroughly. The method using naphthol--ASBI phosphoric acid--fast garnet GBC is recommended for this purpose, and factors crucial to the cytochemical study, such as fixation, substrate, coupler, pH and temperature of incubation buffer, counterstains, and mounting media are examined and discussed. Conventional methods for acid phosphatase in the presence and absence of L(+) tartaric acid are also critically examined. The naphthol--ASBI phosphoric acid--fast garnet GBC method is sensitive, technically simple and easily reproducible. Its reaction product is highly chromogenic and is most suitable for cytochemical demonstration of acid phosphatase and tartrate-resistant acid phosphatase activity in cytologic preparations. The naphthol--ASBI phosphoric acid--pararosaniline method is highly specific and is best for histochemical demonstration of acid phosphatase and tartrate-resistant acid phosphatase in tissue sections.

Topics: Acid Phosphatase; Buffers; Glycerophosphates; Histocytochemistry; Hydrogen-Ion Concentration; Indicators and Reagents; Leukemia, Hairy Cell; Naphthalenes; Naphthols; Organophosphorus Compounds; Phosphoric Acids; Staining and Labeling; Tartrates; Temperature; Time Factors

Immunohematologic studies on cells from a patient with the clinicopathologic syndrome of hairy-cell leukemia showed that the neoplastic cells had receptors for sheep erythrocytes and therefore had human T-lymphocyte characteristics. The leukemic cells did not have the membrane receptors or immunoglobulin markers of B lymphocytes or monocytes nor did they synthesize immunoglobulin. A lymphoid cell line established in vitro from the cells had the same T-lymphocyte characteristics. The lymphoid cell line is positive for tartrateresistant acid phosphatase, forms rosettes with untreated sheep erythrocytes, and reacts with an anti-T-lymphocyte antiserum. Thus the syndrome of hairy-cell leukemia may occasionally result from the neoplastic proliferation of T-lymphocytes as well as from the more usual B-lymphocyte form. This situation is analogous to that described previously in chronic lymphocytic leukemia and other lymphoproliferative disorders.

Topics: Acid Phosphatase; Adult; Animals; Cell Line; Erythrocytes; Humans; Immunoglobulins; Leukemia, Hairy Cell; Male; Rosette Formation; Sheep; T-Lymphocytes; Tartrates

Topics: Acid Phosphatase; B-Lymphocytes; Humans; Immunoglobulin Fc Fragments; In Vitro Techniques; Leukemia, Hairy Cell; Monocytes; Phagocytosis; Receptors, Antigen, B-Cell

The immunobiologic characteristics of three continuous cell lines established from hairy cell leukemia cells were investigated. All three cell lines continued to produce tartrate-resistant acid phosphatase, the enzymatic marker of hairy cells. Two of these cell lines were B lymphoid in nature. They carried Fc and C receptors, had surface and internal immunoglobulin, and did not form spontaneous sheep red blood cell rosettes. Experiments employing biosynthetic radiolabeling of immunoglobulin demonstrated distinctive immunoglobulin kinetics for each of these two hairy cell lines. One cell line remained quite similar to the original hairy cells from which it was derived whereas the other B lymphoid hairy cell line had undergone a switch in the immunoglobulin isotype produced. The third hairy cell leukemia line was shown to be of thymic derivation. These cells formed spontaneous sheep red blood cell rosettes and did not carry Fc or C receptors. The spontaneous sheep red blood cell rosette-forming cells contained tartrate-resistant acid phosphatase. They did not possess surface on internal immunoglobulin and did not synthesize immunoglobulin in vitro. Hairy cell leukemia cells maintained in permanent cell culture retain their immunobiologic properties and offer the opportunity for indepth study of these unusual cells.

Topics: Acid Phosphatase; beta 2-Microglobulin; Binding Sites, Antibody; Cell Line; Cells, Cultured; Cytoplasm; Humans; Immunoglobulin Fc Fragments; Immunoglobulin G; Immunoglobulin M; Leukemia, Hairy Cell; Rosette Formation; Tartrates; Time Factors

Topics: Acid Phosphatase; Cell Count; Complement System Proteins; DNA, Neoplasm; Humans; Immunoglobulins; In Vitro Techniques; Leukemia, Hairy Cell; Macrophages; Microscopy, Electron, Scanning; Phagocytosis; Receptors, Antigen, B-Cell; Rosette Formation; Spleen; Thymidine

A tartrate-resistant acid phosphatase was isolated from a huma spleen infiltrated with reticulum cells of leukemic reticuloendotheliosis. The purified enzyme was used to establish the optimal conditions for quantitative analysis of this enzyme by electrophoresis on acrylamide gel. As little as 0.1 unit of enzyme activity (or 1 ng of the purified enzyme protein) could be quantitatively detected when it was mixed with 4 mg of albumin before electrophoresis.

Topics: Acid Phosphatase; Electrophoresis, Polyacrylamide Gel; Humans; Leukemia, Hairy Cell; Osmolar Concentration; Serum Albumin, Bovine; Spleen; Tartrates; Time Factors

A distinctive pattern of alpha-naphthyl butyrate esterase positivity was observed in all of a series of 14 patients with HCL. This pattern was not observed in a range of other haematological malignancies including nine cases of CLL, two cases of Sézary's syndrome, six cases of null cell ALL, two cases of Schilling-type monocytic leukaemia and one case of lymphosarcoma cell leukaemia. The potential diagnostic value of this simple cytochemical test is discussed in relation to existing methods.

Topics: Acid Phosphatase; Clinical Enzyme Tests; Esterases; Humans; Leukemia, Hairy Cell; Lymphocytes; Rosette Formation

Topics: Acid Phosphatase; Bone Marrow; Humans; Isoenzymes; Leukemia, Hairy Cell; Primary Myelofibrosis

Report on a now 58-year-old female patient who presented in 1969 with the following objective findings: anemia, thrombocytopenia, lymphocytosis and hepatosplenomegaly. The first first diagnosis was lymphosarcoma, but up to 1976 nearly ten other diagnoses were established, e.g. reticulum sarcoma, leukemic lymphosarcoma, CML, ALL, CLL and Waldenström's disease. The patient did not respond to combined chemotherapy, but splenectomy brought about significant improvement in anemia and thrombocytopenia. She is now doing well without cytotoxic treatment. Hairy cell leukemia was diagnosed on the basis of electron microscopic findings in peripheral lymphocytes and histologic findings in bone marrow and spleen. The tartrate-resistant acid phosphatase was positive. Immunologic tests in the hairy cells showed surface immunoglobulins of monoclonal origina and the T-cells were shown to be decreased. The tests for phagocytosis and nonspecific esterase were negative. The hairy cells in this patient exhibited no colony stimulating activity in culture studies with bone marrow from 4 normal donors, as compared with the stimulating activity of normal monocytes in the same assay. In the light of this case report the question is discussed whether the hairy cells are mainly cells with lymphocytic (B-lymphocytes) or monocytic characteristics. On this question the findings in our patient support the hypothesis of a B-cell nature for the hairy cells.

Topics: Acid Phosphatase; Cell Division; Female; Humans; Leukemia, Hairy Cell; Lymphocytes; Middle Aged; Monocytes; Phagocytosis; Receptors, Antigen, B-Cell

Topics: Acid Phosphatase; Adenosine Triphosphatases; Adult; Aged; Alkaline Phosphatase; Esterases; Female; Glucuronidase; Humans; Immunoenzyme Techniques; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphoma; Lymphoma, Large B-Cell, Diffuse; Male; Middle Aged; Mycosis Fungoides; Nucleotidases; Rosette Formation

Four cases of malignant histiocytosis with leukemic manifestations and chronic course were reported. Light microscopic, ultrastructural and ultrastructural cytochemical details of these atypical cells were demonstrated. Ultrastructurally these cells resembled hairy cells most closely among the known varieties of leukemic cells. However, ribosome-lamella complexes were not found and some atypical cells had a few short cytoplasmic projections. In addition, tartrate-resistant acid phosphatase was absent from these cells. We speculate that this leukemic reticuloendotheliosis with a chronic course seen in Japan seems to be analogous to malignant histiocytosis with massive splenomegaly reported by Vardiman et al.

Topics: Acid Phosphatase; Adolescent; Adult; Blood Cells; Bone Marrow; Cytoplasm; Female; Humans; Leukemia, Hairy Cell; Liver; Lymph Nodes; Lysosomes; Male; Ribosomes; Spleen

The activities of acid phosphatases (AP) were measured in leukocytes from patients with chronic myelocytic leukemia (CML), macrophages, granulocytes, in the fractionated mononuclear cells of patients with CML and with hairy-cell-leukemia (HCL) and in the cells from patients with acute leukemia (AL). The lowest activities were found in lymphocytes of normal subjects and of patients with chronic lymphatic leukemia (CLL) and in thrombocytes. Isoenzyme (IsE) 1 was characteristic for thymocytes, IsE 2 for granulocytes, IsE 3 for pathologic blast-cells, lymphocytes and thrombocytes, IsE 4 for macrophages, IsE 5 with components a and b for the mononuclear fraction of patients with HCL. In addition IsE 5 was detected in lymphocytes, macrophages and CLL-cells. In 4 patients with HCL the relative percentage of IsE-5-fraction was slightly greater than the percentage of tartrate resistant cells. In two patients with questionable HCL well marked IsE-5-fractions were recognized but no tartrate resistant cells. In one patient with HCL a relatively high percentage of tartrate resistant hairy-cells and in comparison an inadaquate low IsE-5-fraction was found. These different relations were explained with the more sensitive method of gelelectrophoresis and different affinity of substrates to AP.

Topics: Acid Phosphatase; Acute Disease; Blood Platelets; Granulocytes; Humans; Isoenzymes; Leukemia; Leukemia, Hairy Cell; Leukemia, Myeloid; Leukocytes; Lymphocytes; Macrophages

Topics: Acid Phosphatase; Adolescent; Adult; B-Lymphocytes; Child; Female; Humans; Leukemia, Hairy Cell; Leukemia, Lymphoid; Lymphoma; Male; Periodic Acid-Schiff Reaction; Prognosis; Receptors, Antigen, B-Cell; Rosette Formation; T-Lymphocytes