scholarly journals Flow cytometry of blood and bone marrow cells from patients with hairy cell leukemia: phenotype of hairy cells and lymphocyte subsets after treatment with 2-chlorodeoxyadenosine

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3672-3681 ◽  
Author(s):  
G Juliusson ◽  
R Lenkei ◽  
J Liliemark
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Nk Cells ◽  
Hairy Cell Leukemia ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Cd4 Cells ◽  
Normal Range ◽  
Hla Dr ◽  
Hairy Cells

Abstract By flow cytometry and an extensive set of markers, we characterized leukemic cells from the blood and bone marrow of 68 symptomatic patients with hairy cell leukemia (HCL). Hairy cells identified in the large cell gate always expressed CD19, CD20, HLA-DR, CD45RA, and B-ly 7. Other markers were occasionally expressed, such as CD38, CD45RO, CD23, CD15, CD4, CD5, and CD10 (expressed on more than 20% of the hairy cells in 44%, 25%, 21%, 18%, 12%, 10%, and 5% of evaluated cases, respectively). During treatment with 2-chlorodeoxyadenosine (CdA), the median lymphocyte counts decreased from 2,000/microL to 300/microL. Flow cytometry was repeated at the nadir (n = 24) of lymphocyte counts, at 3 months (n = 46), at 6 months (n = 50), at 1 year (n = 39), and at 2 years (n = 12) after treatment. The initial decrease of CD8+ and CD20+ cells was greater than that of CD4+ and natural killer (NK) cells, leading to an increasing CD4/CD8 ratio. Median nadir values of CD4+, CD8+, CD20+, and NK cells were 128/microL, 78/microL, 10/microL, and 13/microL, respectively. The subsequent recovery was quicker for CD8+ and NK cells, leading to a normalization within 3 months, whereas CD20+ and CD4+ cells required 1 or 2 years to enter the normal range. The CD4/CD8 ratio thus decreased after the nadir and remained less than 1. CD45RA+ CD4 cells and CD45RA+/CD45RO+ double-positive cells were less affected by CdA. Activated T cells, ie, HLA-DR+ cells, rarely decreased below the normal range and often recovered with an overshoot. CD10+ cells increased in the bone marrow posttreatment as an indication of normal B-cell regeneration in 16 of 36 (44%) patients. The quick regeneration of certain lymphoid subsets might explain the lack of late infections in CdA-treated HCL patients.

scholarly journals Flow cytometry of blood and bone marrow cells from patients with hairy cell leukemia: phenotype of hairy cells and lymphocyte subsets after treatment with 2-chlorodeoxyadenosine

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3672-3681 ◽  
Author(s):  
G Juliusson ◽  
R Lenkei ◽  
J Liliemark
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Nk Cells ◽  
Hairy Cell Leukemia ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Cd4 Cells ◽  
Normal Range ◽  
Hla Dr ◽  
Hairy Cells

By flow cytometry and an extensive set of markers, we characterized leukemic cells from the blood and bone marrow of 68 symptomatic patients with hairy cell leukemia (HCL). Hairy cells identified in the large cell gate always expressed CD19, CD20, HLA-DR, CD45RA, and B-ly 7. Other markers were occasionally expressed, such as CD38, CD45RO, CD23, CD15, CD4, CD5, and CD10 (expressed on more than 20% of the hairy cells in 44%, 25%, 21%, 18%, 12%, 10%, and 5% of evaluated cases, respectively). During treatment with 2-chlorodeoxyadenosine (CdA), the median lymphocyte counts decreased from 2,000/microL to 300/microL. Flow cytometry was repeated at the nadir (n = 24) of lymphocyte counts, at 3 months (n = 46), at 6 months (n = 50), at 1 year (n = 39), and at 2 years (n = 12) after treatment. The initial decrease of CD8+ and CD20+ cells was greater than that of CD4+ and natural killer (NK) cells, leading to an increasing CD4/CD8 ratio. Median nadir values of CD4+, CD8+, CD20+, and NK cells were 128/microL, 78/microL, 10/microL, and 13/microL, respectively. The subsequent recovery was quicker for CD8+ and NK cells, leading to a normalization within 3 months, whereas CD20+ and CD4+ cells required 1 or 2 years to enter the normal range. The CD4/CD8 ratio thus decreased after the nadir and remained less than 1. CD45RA+ CD4 cells and CD45RA+/CD45RO+ double-positive cells were less affected by CdA. Activated T cells, ie, HLA-DR+ cells, rarely decreased below the normal range and often recovered with an overshoot. CD10+ cells increased in the bone marrow posttreatment as an indication of normal B-cell regeneration in 16 of 36 (44%) patients. The quick regeneration of certain lymphoid subsets might explain the lack of late infections in CdA-treated HCL patients.

scholarly journals Low-dose deoxycoformycin in the treatment of hairy cell leukemia

Blood ◽  
1986 ◽  
Vol 68 (5) ◽  
pp. 1119-1122 ◽  
Author(s):  
EH Kraut ◽  
BA Bouroncle ◽  
MR Grever
Keyword(s):  
Bone Marrow ◽  
Complete Remission ◽  
Hairy Cell Leukemia ◽  
Low Dose ◽  
Intravenous Bolus ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Normal Peripheral Blood ◽  
Hairy Cells ◽  
Reversible Reduction

Abstract Ten patients with progressive hairy cell leukemia were treated with 2′deoxycoformycin (dCF) by intravenous bolus (4 mg/m2) given every other week. All ten patients are evaluable for response and nine of the ten patients have achieved a complete remission. In addition to clearing of hairy cells from the bone marrow, eight patients had resolution of their monocytopenia. Seven of the nine patients remain in unmaintained remission with a median duration of 6.2 months. Two patients have had relapse in the bone marrow alone and continue to have normal peripheral blood counts. They are being followed without treatment. Toxicity was minimal at this low dose with one patient having a mild reversible reduction in creatinine clearance. Four other patients had reversible neutropenia. There were no significant infections associated with treatment. Low-dose deoxycoformycin administered intravenously every other week represents an extremely effective treatment for hairy cell leukemia.

Treatment of hairy cell leukemia with alternating cycles of pentostatin and recombinant leukocyte A interferon: results of a phase II study.

1990 ◽  
Vol 8 (4) ◽  
pp. 721-730 ◽  
Author(s):  
A Martin ◽  
S Nerenstone ◽  
W J Urba ◽  
D L Longo ◽  
J B Lawrence ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Hairy Cell Leukemia ◽  
Pathologic Complete Response ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Cell Infiltrate ◽  
Recombinant Interferon ◽  
Bone Marrow Biopsies ◽  
Hairy Cells

Fifteen patients with hairy cell leukemia (HCL) were treated with deoxycoformycin (pentostatin; dCF) (4 mg/m2 intravenous [IV] every week x 3) and recombinant interferon-alpha 2a (rIFN-alpha 2a) (3 x 10(6) units subcutaneously [SC] daily x 4 weeks) in alternating months for a total of 14 months. Eleven patients had undergone splenectomy; four had received prior systemic therapy with chlorambucil and/or steroids. All 15 are evaluable for toxicity and peripheral blood response, while 14 are assessable for bone marrow response. Toxicity was tolerable with grade 3 or 4 nausea and vomiting in three patients, neutropenic fevers in five, transient but significant depression in eight, and localized cutaneous herpes zoster in four. Circulating hairy cells were undetectable by the end of the first month in 10 of 13 patients, and by the end of the second month in the other three. Fourteen patients had bilateral bone marrow biopsies performed at baseline after 6 months of treatment, at the end of treatment (14 months), and at 6-month intervals during follow-up. Before treatment, all patients had hypercellular marrows with hairy cels replacing normal marrow elements; all showed at least a 95% clearing of their hairy cell infiltrate by 6 months of therapy. However, small collections of residual hairy cells could be detected intermittently on at least one side of bilateral samples in all patients. All patients have completed treatment with a median duration of follow-up off therapy of 27 months (range, 15 to 31 months). To date, all peripheral counts and serum soluble interleukin-2 receptor (sIL2R) levels remain stable, and no patient has had progression of the hairy cell infiltrate in the bone marrow. Although no patient achieved a pathologic complete response, alternating monthly cycles of dCF and rIFN-alpha 2a produced durable partial remissions (PRs) in all patients. Continued follow-up is required to determine the length of such remissions.

Hairy-cell leukemia: induction of complete remission with pentostatin (2'-deoxycoformycin).

1984 ◽  
Vol 2 (12) ◽  
pp. 1336-1342 ◽  
Author(s):  
A S Spiers ◽  
S J Parekh ◽  
M B Bishop
Keyword(s):  
Bone Marrow ◽  
Complete Remission ◽  
Hairy Cell Leukemia ◽  
Bone Marrow Aspiration ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Highly Active ◽  
Extensive Evaluation ◽  
Ann Arbor ◽  
Hairy Cells

Two men with advanced but previously untreated B cell hairy-cell leukemia were treated with low doses of pentostatin (2'-deoxycoformycin) in intermittent courses. There was prompt clearance of hairy cells from the blood, regression of splenomegaly and lymphadenopathy, and correction of anemia, thrombocytopenia, and granulocytopenia. Side effects were tolerable and myelosuppression was not observed. Both patients achieved complete remission documented by bone marrow aspiration and biopsy and radionuclide scans of liver and spleen. They remain in complete remission nine and six months, respectively, after their last treatment. Pentostatin (Warner-Lambert, Ann Arbor, Mich) is highly active in hairy-cell leukemia and merits more extensive evaluation in this disease. A woman with hairy-cell leukemia has begun treatment with pentostatin, and at ten weeks there is disappearance of gross splenomegaly and clearance of hairy cells from the blood. Bone marrow studies have not yet been repeated.

scholarly journals Usefulness of Dual Immunohistochemistry Staining in Detection of Hairy Cell Leukemia in Bone Marrow

2019 ◽  
Vol 153 (3) ◽  
pp. 322-327 ◽  
Author(s):  
Gaurav K Gupta ◽  
Xiaoping Sun ◽  
Constance M Yuan ◽  
Maryalice Stetler-Stevenson ◽  
Robert J Kreitman ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Hairy Cell Leukemia ◽  
Predictive Value ◽  
Lymphoid Cells ◽  
Multiparameter Flow Cytometry ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Flow Cytometry Data ◽  
Flow Cytometric Data

Abstract Objectives We evaluated efficacy of two dual immunohistochemistry (IHC) staining assays in assessing hairy cell leukemia (HCL) involvement in core biopsies and compared the results with concurrently collected flow cytometric data. Methods Overall, 148 patients with HCL (123 male, 25 female; mean age: 59.8 years; range: 25-81 years) had multiparameter flow cytometry performed using CD19, CD20, CD22, CD11c, CD25, CD103, CD123, surface light chains, CD5, and CD23. In parallel, bone marrow IHC was done using PAX5/CD103 and PAX5/tartrate-resistant alkaline phosphatase (TRAP) dual IHC stains. Results Overall sensitivity of dual IHC stains was 81.4%, positive predictive value was 100%, and negative predictive value was 81.7%. All IHC-positive cases concurred with flow cytometry data, even when HCL burden was extremely low in the flow cytometry specimens (as low as 0.02% of all lymphoid cells). Conclusions Dual IHC stain is a sensitive tool in detecting HCL, even in cases with minimal disease involvement.

scholarly journals Treatment of hairy cell leukemia with recombinant alpha interferon: I. Quantitative study of bone marrow changes during the first months of treatment

Blood ◽  
1986 ◽  
Vol 67 (3) ◽  
pp. 817-820 ◽  
Author(s):  
G Flandrin ◽  
F Sigaux ◽  
S Castaigne ◽  
C Billard ◽  
M Aguet ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Hairy Cell Leukemia ◽  
Alpha Interferon ◽  
Initial Increase ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Bone Marrow Biopsies ◽  
Biopsy Specimens ◽  
Hairy Cells ◽  
Normal Controls

Abstract Seventeen patients with hairy cell leukemia (HCL) were treated with low doses of recombinant alpha interferon (IFN) for over 4 months. Marked improvement was observed in peripheral blood and bone marrow in 15 of 17 patients. Comparison of pretreatment values and hemograms obtained after 4 months of treatment showed a marked decrease in circulating hairy cells (P less than .01), a decrease in the number of lymphocytes (P less than .01), a rise in the number of platelets (P less than .05), granulocytes (P less than .05), and monocytes (P less than .01), and a rise in the hemoglobin level (P less than .01). Transient reduction in the number of granulocytes was noted during the first month. Correction of thrombocytopenia often appeared within 2 months and usually preceded improvement of anemia, monocytopenia, and neutropenia. Bone marrow biopsy specimens were taken before treatment and 2, 4, and 7 months after its initiation. The volumes occupied by hairy cells, cells of the myeloid lines, and adipocytes were studied by stereological analysis of semithin sections. Decrease in the volume occupied by hairy cells was seen after 4 months of treatment (P less than .01), and the volume continued to decrease at the seventh month (P less than .05). Hairy cells were no longer detected on bone marrow biopsies of 4 of 17 patients by the fourth month and in 3 of 8 additional patients by the seventh month. A rise in the volume occupied by normal myeloid cells was visible by the second month of treatment (P less than .01). Nevertheless, the volume occupied by granulocytes remained lower than in the normal controls (P less than .01). After an initial increase during the first 2 months of treatment (P less than .01), the overall cellularity remained unchanged at 4 months and decreased significantly (P less than .05) at 7 months. Except for biopsies at 2 months, mean cellularity was below that of control biopsies (P less than .01).

scholarly journals Morphologic confounders and CD19 negativity in a case of hairy cell leukemia.

2017 ◽  
Vol 9 (1) ◽  
pp. e2017033
Author(s):  
Pulkit Rastogi ◽  
Sreejesh Sreedharanunni ◽  
Uday Yanamandra ◽  
Man Updesh Singh Sachdeva ◽  
Neelam Varma
Keyword(s):  
Bone Marrow ◽  
Hairy Cell Leukemia ◽  
Plasma Cell Dyscrasia ◽  
Molecular Testing ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Cell Hyperplasia ◽  
Reactive Plasmacytosis ◽  
Cd10 Expression ◽  
Hairy Cells

Objectives:We report a case of hairy cell leukemia (HCL) initially misdiagnosed as plasma cell dyscrasia due to various clinical, morphological and immunophenotypic confounders.Methods and results:In a patient diagnosed of marrow plasmacytosis and serum monoclonal protein elsewhere and referred to our hospital, morphological evaluation of bone marrow aspirate smears and trephine biopsy, immunophenotyping, and molecular testing (BRAFV600E mutation) were done. Clinically, the patient was asymptomatic, bone marrow revealed plasmacytosis, mastocytosis and lymphocytosis with a few “hairy” cells. Immunophenotyping revealed features of HCL with aberrant CD10 expression and a subclone of CD19neg cells. A diagnosis of HCL with reactive plasmacytosis and mast cell hyperplasia was made and confirmed by immunophenotyping and molecular studies.Conclusion:Hematopathologists must be aware of various confounding factors and should judiciously use flow cytometric and molecular studies for attaining a proper diagnosis of HCL. We also report a very rare immunophenotypic aberrancy (CD 19 negativity) in HCL

scholarly journals Diagnostic application of two-color flow cytometry in 161 cases of hairy cell leukemia

Blood ◽  
1993 ◽  
Vol 82 (4) ◽  
pp. 1277-1287 ◽  
Author(s):  
BA Robbins ◽  
DJ Ellison ◽  
JC Spinosa ◽  
CA Carey ◽  
RJ Lukes ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Hairy Cell Leukemia ◽  
Cell Phenotype ◽  
Specific Marker ◽  
Positive Staining ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Color Flow ◽  
Intense Staining ◽  
Hairy Cells

Recent immunophenotypic studies of hairy cell leukemia (HCL) have suggested specific patterns of immunoreactivity that may aid in diagnosis. We studied peripheral blood (PB) from 161 cases of HCL using two-color direct immunofluorescence flow cytometry and an extended panel of antibody combinations. Circulating hairy cells were identified by immunophenotypic features in 92% of the cases and could be detected even when representing < or = 1% of circulating lymphocytes. The 133 cases with > or = 2% detectable hairy cells were analyzed in detail. HCL showed a uniform and unique B-cell phenotype, with each of the following features identified in 99% to 100% of cases: (1) positive staining for B-ly7, coexpressed with CD19; (2) very intense, uniform expression of CD11c, with CD19; (3) moderately intense staining for CD25, with CD19; (4) very intense staining for CD22; (5) moderate to very intense staining for CD20; and (6) moderately intense monoclonal surface Ig. Phenotypic variability existed in expression of CD10 (26%) and CD5 (4%). Based on these features, HCL was easily distinguished from 50 cases of chronic lymphocytic leukemia (CLL). Although CLL exhibited frequent expression of CD11c (74%) and CD25 (68%), the intensity of staining was significantly less than HCL. Furthermore, CLL was uniformly positive for CD5 and showed weak staining for CD20, CD22, and surface Ig. B-ly7 proved to be the most specific marker, reacting with 100% of HCL cases, but absent in all cases of CLL. We conclude that two-color flow cytometry with specific antibody combinations is an efficacious method for characterization and sensitive detection of hairy cells in PB. Application of the phenotypic criteria described should help to increase accuracy in diagnosis of HCL.

scholarly journals The Role of Novel Dual Color Immunohistochemistry in Detection of Minimal Hairy Cell Leukemia in Bone Marrow: A Study of 148 Cases

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 4859-4859
Author(s):  
Gaurav K. Gupta ◽  
Xiaoping Sun ◽  
Constance M. Yuan ◽  
Maryalice Stetler-Stevenson ◽  
Robert J. Kreitman ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Hairy Cell Leukemia ◽  
Predictive Value ◽  
Lymphoid Cells ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Flow Cytometry Data ◽  
Flow Cytometric ◽  
Minimal Disease

Abstract Background: Hairy-cell leukemia (HCL) is a B-cell lymphoproliferative disorder characterized by distinct immunophenotype (positive for CD19, CD20, PAX5, CD22, CD11c, CD25, CD103, CD123 and CD200). Both flow cytometry (FC) and immunohistochemistry (IHC) can be used to determine these markers, while the expression of another marker (TRAP) is IHC specific. Both trephine bone marrow biopsy and aspirate are vital for assessment of extent of bone marrow infiltration. However, in some cases a cellular aspirate cannot be obtained due to extensive fibrosis, ie "dry tap". In such cases, IHC stains are crucial for assessment of bone marrow leukemic involvement. So far, IHC detection of HCL in the bone marrow sections has been limited to overt disease and could not be reliably used in cases with minimal HCL involvement. Novel automated dual-antibody immunohistochemistry techniques can identify aberrant antigen co-expression in neoplastic cells with high sensitivity. Consistent detection of minimal disease involvement is crucial given that treatment decisions are sometimes based on these data (Grever et. al. Blood 2017). In this study, we evaluated the efficacy of two novel dual IHC assays in assessing minimal HCL involvement in the core biopsies and compared the results with concurrently collected flow cytometric data. Design: We analyzed data on 148 cases of HCL (123 male, 25 female; mean age 59.8, range 25-81). All cases had multiparameter flow cytometry performed using CD19, CD20, CD22, CD11c, CD25, CD103, CD123, surface light chains, CD5 and CD23. In parallel, bone marrow IHC was done using PAX5/CD103 and PAX5/TRAP dual stains and the automated stainer (Ventana Ultra). Results: Cases were divided into three groups based on the combined results of PAX5/CD103 and PAX5/TRAP stains: negative (82; 55.4%); rare dual positive cells (less than 5% of total cells) (21; 14.1%) and positive (45; 30.4%). Flow cytometry data concurred with IHC results in all IHC positive cases (median 7.05% HCL cells out of all lymphoid cells by FC; range 0.05%-91.7%) and all rare dual-cell IHC positive cases (median 0.98% HCL cells; range 0.02%-19.04%). Overall sensitivity of dual IHC was 81.4%, positive predictive value 100% and negative predictive value 81.7%. In dual IHC negative group, 15/82 cases (18.3%) were low level positive by FC analysis (median 0.13% HCL cells; range 0.01%-9.21%). When dual IHC results were analyzed separately, PAX5/CD103 results were similar to the combined results. PAX5/TRAP staining alone was slightly less sensitive in IHC negative cases; 22/82 (26.8%) of PAX5/TRAP negative or non-evaluable cases were positive by FC analysis (median 0.27% HCL cells, range 0.01%-29.5%). Conclusion: Dual color IHC is a sensitive tool in detecting HCL, even in cases with minimal disease involvement. All IHC positive cases concurred with flow cytometry data, even when HCL burden was extremely low (as low as 0.02% of all lymphoid cells by flow cytometric analysis). Only 18.3% of dual IHC negative cases were positive for low level involvement by FC analysis. Therefore, dual IHC is a sensitive new tool for evaluation of minimal marrow involvement by HCL. Figure. Figure. Disclosures Kreitman: NIH: Patents & Royalties: Co-inventor on the NIH patent for Moxetumomab Pasudotox.

scholarly journals Diagnostic application of two-color flow cytometry in 161 cases of hairy cell leukemia

Blood ◽  
1993 ◽  
Vol 82 (4) ◽  
pp. 1277-1287 ◽  
Author(s):  
BA Robbins ◽  
DJ Ellison ◽  
JC Spinosa ◽  
CA Carey ◽  
RJ Lukes ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Hairy Cell Leukemia ◽  
Cell Phenotype ◽  
Specific Marker ◽  
Positive Staining ◽  
Hairy Cell ◽  
Cell Leukemia ◽  
Color Flow ◽  
Intense Staining ◽  
Hairy Cells

Abstract Recent immunophenotypic studies of hairy cell leukemia (HCL) have suggested specific patterns of immunoreactivity that may aid in diagnosis. We studied peripheral blood (PB) from 161 cases of HCL using two-color direct immunofluorescence flow cytometry and an extended panel of antibody combinations. Circulating hairy cells were identified by immunophenotypic features in 92% of the cases and could be detected even when representing < or = 1% of circulating lymphocytes. The 133 cases with > or = 2% detectable hairy cells were analyzed in detail. HCL showed a uniform and unique B-cell phenotype, with each of the following features identified in 99% to 100% of cases: (1) positive staining for B-ly7, coexpressed with CD19; (2) very intense, uniform expression of CD11c, with CD19; (3) moderately intense staining for CD25, with CD19; (4) very intense staining for CD22; (5) moderate to very intense staining for CD20; and (6) moderately intense monoclonal surface Ig. Phenotypic variability existed in expression of CD10 (26%) and CD5 (4%). Based on these features, HCL was easily distinguished from 50 cases of chronic lymphocytic leukemia (CLL). Although CLL exhibited frequent expression of CD11c (74%) and CD25 (68%), the intensity of staining was significantly less than HCL. Furthermore, CLL was uniformly positive for CD5 and showed weak staining for CD20, CD22, and surface Ig. B-ly7 proved to be the most specific marker, reacting with 100% of HCL cases, but absent in all cases of CLL. We conclude that two-color flow cytometry with specific antibody combinations is an efficacious method for characterization and sensitive detection of hairy cells in PB. Application of the phenotypic criteria described should help to increase accuracy in diagnosis of HCL.

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