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Results for “Immunophenotyping”

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PubMed1995

Immunophenotyping of B lymphocytes by multiparametric flow cytometry in bone marrow aspirates and peripheral blood of healthy adults.

Establishing reference ranges by multiparametric immunophenotyping of mature B cells in bone marrow and peripheral blood of healthy adults is of interest because the detection of bone marrow infiltration, persistance of light chain restriction as well as discrimination between reactice and malignant lymphocytes are important applications of B-cell immunophenotyping. To determine the pattern of antigens as expressed by malignant mature B lymphocytes, bone marrow aspirates and peripheral blood of healthy adults in the present study were investigated for the presence and percentage frequency of those antigens as defined for immunophenotyping of B-cells by the REAL-Classification. For this purpose analysis of CD19 positive B lymphocytes by Live Gate analysis was performed. The established two-color as well as three-color stainings will serve as a basis for future investigations designed to test multiparametric analysis of B lymphocytes in bone marrow aspirates and peripheral blood. In conclusion, all investigated antibodies stained in varying percentage frequency on B-cell subtypes and statistical significant differences could be considered only for the CD19/CD10 staining in bone marrow aspirates. On the basis of this analysis, all the reported lineage antigen combinations are present both in malignant B lymphocytes as well as normal B cells in considerable percentage frequency. These findings are of important interest for follow-up investigations of patients with non-Hodgkin s lymphomas by multiparametric immunophenotyping.

Adult↗

PubMedDate not supplied

[Cellular immunophenotypes in 97 adults with acute leukemia].

OBJECTIVE: To analyze hematopoietic cell surface antigen reactivity in acute leukemia (AL) by flow cytometry and identify acute mixed-lineage leukemias (AMLL) employing the most widely accepted criteria. MATERIAL AND METHODS: Ninety seven patients with de novo AL were studied. Cell surface antigens were investigated with monoclonal antibodies directed to: B lymphoid (CD10, CD19, CD20, CD21, CD22); T lymphoid (CD2, CD3, CD5, CD7); and myeloid (CD13, CD14, CD15, CD33, CD41) cell lineages. Maturation cell-associated antigens (CD34, HLA-DR and TdT) were also studied. RESULTS: Twelve patients unclassified by cytomorphology could be classified by immunophenotype. Using cytomorphologic, cytochemical and immunophenotypic data, 54 cases corresponded to acute lymphoblastic leukemia (ALL) and 43 were acute myeloblastic leukemia (AML). In All there were 63% B lineage, 15% T, 7% T/B, 6% undifferentiated and 9% mixed-lineage (coexpression of two or more myeloid-associated antigens). In AML, myeloid immunophenotype was observed in 86% undifferentiated in 2%, and mixed-lineage in 12% (coexpression of two or more lymphoid-associated antigens). In addition, 26% of ALL cases and 12% of AML cases expressed a single myeloid and lymphoid antigen respectively. The most common aberrant antigens in ALL and AML were CD13 and CD7 respectively. The highest frequency of CD34 antigen expression (90%) was detected in patients with AMLL. CONCLUSIONS: Flow cytometric immunophenotypic analysis allowed to: a) establish diagnosis in cytomorphologically unclassified cases; b) identify AMLL with a frequency similar to that reported in other series; and c) confirm the heterogeneity of AL.

Acute Disease↗

PubMed1998

Indolent systemic mast cell disease in adults: immunophenotypic characterization of bone marrow mast cells and its diagnostic implications.

The aim of the present study was to explore the diagnostic value of the immunophenotypic analysis of bone marrow mast cells (BMMC) in indolent systemic mast cell disease (SMCD) patients. For that purpose, a total of 10 SMCD patients and 19 healthy controls were analyzed. Our results show that BMMC from SMCD are different from normal BMMC with regard to both their light scatter and immunophenotypic characteristics. Accordingly, forward light scatter (FSC), side (90 degrees) light scatter (SSC), and baseline autofluorescence levels were higher in BMMC from indolent SMCD patients than they were in control subjects. From the immunophenotypic point of view, the most striking findings were the constant expression of CD2 (P = .0001), CD25 (P = .0001), and CD35 (P = .06) molecules by BMMC from SMCD patients, markers that were absent from all normal controls. In contrast, CD71, absent in BMMC from indolent SMCD, was positive in BMMC from normal subjects. Although, slight differences between BMMC from SMCD patients and normal controls were found in several other markers, they did not reach statistical significance. In conclusion, our results show that simultaneous assessment of FSC/SSC and reactivity for the CD117, CD2, CD25, CD33, and CD35 forms the basis for the immunophenotypic characterization of BMMC from SMCD in adults and should be integrated with clinical and morphologic studies for the diagnosis of the disease.

Adult↗

PubMed2006

Cutaneous lymphoid hyperplasia presenting as a solitary facial nodule: clinical, histopathological, immunophenotypical, and molecular studies.

OBJECTIVE: To elucidate the clinicopathological, immunophenotypical, and molecular characteristics of cutaneous lymphoid hyperplasia presenting as a solitary facial nodule. DESIGN: Retrospective study. SETTING: University dermatology department. PATIENTS: Three patients with a solitary facial nodule were studied clinically, histologically, immunophenotypically, and molecularly for T-cell receptor and immunoglobulin heavy chain gene rearrangements. MAIN OUTCOME MEASURES: Histological, immunophenotypical, and molecular characteristics in relation to the clinical outcome. RESULTS: Histologically, dense diffuse lymphocytic infiltrates were present throughout the dermis, occasionally extending into the subcutaneous fat and the epidermis and hair follicles. Small lymphocytes predominated, but in 2 cases there were also medium to large atypical lymphocytes, with some blastlike lymphocytes. The lymphocytic population was mixed with more CD3(+) T cells than CD20(+) B cells, without germinal centers. There were more CD4(+) than CD8(+) cells, and some of the T cells stained for the memory T-cell marker CD45RO. Numerous CD68(+) histiocytes were scattered or formed small aggregates, and in 1 case small granulomas and many scattered S100 protein-positive and CD1a(+)dendritic cells were present. In addition, several polytypic plasma cells, eosinophils, and extravasated erythrocytes were found. Immunostaining for CD10, CD21, CD30, CD56, and BCL6 was negative. The Ki-67 proliferation index was relatively low (5%-10%). Results of the T-cell receptor gene rearrangement studies were positive in 2 cases, 1 of which also harbored clonal B cells. Serologic test results for Borrelia burgdorferi, Borrelia afzelii, and Borrelia garinii were negative in all 3 cases. Two lesions regressed spontaneously after an incisional biopsy, and none of the cases showed recurrence or extracutaneous spread during a follow-up period of 5.0 to 5.5 years. CONCLUSIONS: Cutaneous lymphoid hyperplasia that presents as a solitary facial nodule may share clinical, cytological, immunophenotypical, and molecular features with both benign reactive lymphocytic infiltrates and cutaneous lymphomas, and therefore a careful clinical and therapeutic approach is warranted.

Biopsy↗

PubMed1997

Immunophenotype of human ovarian malignancies (cystadenocarcinomata and mixed müllerian tumor) established in SCID mice.

Human ovarian malignancies from three different patients (histology: two serous cystadenocarcinomata and one mixed Müllerian tumor, homologous type) were successfully serially transplanted intraperitoneally into severe combined immunodeficient (SCID) mice where the tumor cells spread around the peritoneal cavity. If the ascites derived from cystadenocarcinoma cells engrafted in the female genital tract of the SCID mice, they formed cystic tumors resembling remarkably well the original tumors in the patients. Immunohistochemical analysis revealed that the immunophenotype of the patients' original tumor and those grown in SCID mice were similar in the case of the two cystadenocarcinomata; in addition, the marker expression in general was stable during serial transplantation. If distant metastases occurred in the lungs, they immunophenotypically resembled those grown intraperitoneally. In contrast, the cells derived from the mixed Müllerian tumor shifted during serial transplantation from a spindle cell morphology toward a morphology characterized by cuboidal cells. The transition toward a more epithelial phenotype was accompanied by a changed immunophenotype of the tumor cells which became positive for epithelial cell markers such as carcinoembryonic antigens, CA 19-9 and CA 125. Concurrently with this differentiation, the p53 immunophenotype changed from positive to negative, indicating a further mutation in the p53 gene during serial passages.

Animals↗

PubMed1990

Factor analysis of the immunophenotypes of astrocytomas and malignant gliomas: correlations with tumor grade and patient survival.

Our previous investigations correlated the degree of cytogenetic and immunophenotypic heterogeneity of cultured normal glia, astrocytomas and malignant gliomas. The possible significance was suggested by the statistical correlation of individual antigens with diagnosis and patient survival. The present study has established the patterns of covariation of titers of monoclonal antibody reactivity with a panel of cell surface antigens among normal glia (8), astrocytomas (4), anaplastic astrocytomas (12), mixed malignant gliomas (8) and glioblastomas (21). A mean aggregate titer across 43 antigens was computed for each culture and then subtracted from the observed individual titers. Factor analysis was performed to determine a small number of Factors, derived as the weighted average of the 43 mean-adjusted antigens, which accounted for a significant proportion of the covariation of immunophenotypic expression in the sample of 53 cultures. Clusters of antigens were found to independently segregate in their deviation from the aggregate phenotype. Adjusting for age and diagnosis, Factors 1 and 4 correlated with patient survival among recurrent and primary neoplasms, respectively. Factor 2 additionally discriminated between primary and recurrent gliomas. Factor 3 was associated with age at diagnosis. Factors 1 and 2 correlated with the histopathologic grade of glial tumor. Scatter plots of Factor 1 vs. 2 revealed the minimal immunophenotypic diversity of the normal glia. Astrocytomas were similar but not identical. Progressive divergence was evident between the immunophenotypes of anaplastic astrocytomas, mixed gliomas and glioblastomas. These data suggest that qualitative and quantitative differences in antigenic heterogeneity may identify stages in glial tumor progression.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗

PubMed1986

Immunophenotypic transformation from acute undifferentiated leukemia to Burkitt's-like acute lymphoblastic leukemia.

Progression of more differentiated to less differentiated malignant phenotypes has been described infrequently during the natural evolution or at relapse of treated hematopoietic malignancies. This report describes an unusual instance of immunophenotypic transformation from an immunologically undifferentiated acute leukemia to a leukemia that at relapse possessed morphologic and immunologic markers characteristic of a Burkitt's-like acute lymphoblastic leukemia. A 26-year-old man initially presented with pancytopenia and a bone marrow diffusely replaced with blast cells morphologically most consistent with a French-American-British L2 subclassification. The surface immunophenotype of the blasts at diagnosis showed HLA-DR surface antigen but no myeloid, lymphoid, or immunoglobulin determinants. Despite successful induction and ongoing consolidation chemotherapy, the patient had a relapse five months after diagnosis; blast cells at relapse demonstrated marked cytoplasmic vacuolation consistent with a Burkitt's-like L3 acute lymphoblastic transformation. Immunophenotypic analysis revealed the presence of restricted immunoglobulin determinants (mu heavy chain and kappa light chain), as well as two separate B lineage surface determinants (BA-1 and B-1). Immunophenotypic transformations may reflect the presence of either a multiclonal or multipotent leukemic population; documentation of the frequency of such transformations and genomic analysis of the transformed subpopulations may be helpful in furthering the understanding of molecular mechanisms involved in leukemogenesis.

Acute Disease↗

PubMed1998

Histologic and immunophenotypic features prior to transformation in patients with transformed cutaneous T-cell lymphoma: is CD25 expression in skin biopsy samples predictive of large cell transformation in cutaneous T-cell lymphoma?

Large cell transformation (LCT) in cutaneous T-cell lymphoma (CTCL) is estimated to occur up to approximately 20% of patients. This morphologic change is associated with aggressive behavior and shortened survival. Our purpose was to identify morphologic and/or immunophenotypic features in CTCL skin biopsies that are predictive of transformation. We analyzed 12 cases of CTCL (three cases of mycosis fungoides and nine cases of Sezary syndrome) that underwent LCT and for which sequential biopsies before and after transformation were available. Eight of 12 cases had pre-LCT marker studies performed on frozen tissue. The histologic and immunophenotypic features of CTCL and LCT were reviewed. Morphologically, CTCL biopsies showed the following: Pautrier microabscesses (five cases), epidermotropism of single lymphoid cells (three cases), and nonspecific histology (four cases). Immunophenotyping prior to transformation showed aberrant loss of T-cell markers (CD5, CD7, CD5 and CD7) in three of eight cases. CD4:CD8 ratios were increased in all cases. Expression of CD25 by the neoplastic lymphoid cells was detected in six of seven cases. LCT immunophenotyping showed additional loss of CD7 in one of five cases and of CD25 in three of five cases. In conclusion, expression of CD25 in CTCL may identify a subset of patients at risk of undergoing LCT.

Adult↗

PubMed1989

Morphologic and immunophenotypic characterization of primary brain lymphomas using paraffin-embedded tissue.

Primary lymphomas of the brain constitute about 1% of all primary intracranial neoplasms, but recent studies suggest an increasing incidence. Most cases are associated with an immunosuppressed state. We reviewed 29 cases of primary brain lymphoma from the Yorkshire Health Authority Region between 1970 and 1988 and found a striking increase in incidence over this period. No overt evidence of immunosuppression was found in any case. All were non-Hodgkin's in type and were classified morphologically using Kiel criteria and immunophenotypically using a panel of antibodies. Cryo-preserved tissue was available in five cases for parallel immunophenotyping. The majority of tumours were high-grade lymphomas together with three of lymphoplasmacytoid type. Thirteen tumours showed a striking pleomorphic morphology with plasmacytoid features. A reactive, predominantly perivascular monomorphic T-cell population was seen in all tumours. Most tumours were of B-cell lineage. No cases of Hodgkin's disease, T-cell or histiocytic lymphoma were present. Light chain restriction was present in only 46% of cases. The results of tumour immunophenotyping on cryostat sections were comparable with those from paraffin blocks. Our study emphasizes the value of a panel of antibodies reactive in paraffin-embedded tissue, allowing simultaneous evaluation of morphology and immunophenotype, and suitable for small biopsies received from stereotactic procedures.

Adolescent↗

PubMed1998

Immunophenotypic comparison of blood and lymph node from dogs with lymphoma.

Peripheral blood and lymph node tissue from 12 dogs with lymphoma was immunophenotyped. Additionally, the bone marrow was immunophenotyped in 6 dogs. The lymphomas were characterized as B-cell in 11 dogs and T-cell in 1 dog. Immunophenotypic patterns in the peripheral blood and bone marrow were variable. The trend in dogs with B-cell lymphoma was normal to increased percentage of IgG-positive cells, decreased percentage of pan-T-positive cells, decreased percentage of CD4-positive cells, and decreased CD4/CD8 ratio. Simultaneous immunophenotyping of lymph node, blood and bone marrow cannot be recommended routinely without further studies to document its value as an independent prognostic indicator. However, it is potentially useful for tumor staging and monitoring remission, especially in lymphoma patients with a leukemic phase.

Journal Article↗

PubMed1990

Characteristics of CD11c+CD5+ chronic B-cell leukemias and the identification of novel peripheral blood B-cell subsets with chronic lymphoid leukemia immunophenotypes.

Previous studies have indicated that chronic lymphocytic leukemias (CLL) are characterized by the coexpression of CD5 and B-cell antigens, while hairy cell leukemias (HCL) typically express CD11c+CD5- B-cell immunophenotypes. In this report we describe the features of B-cell leukemias with CD11c+CD5+ immunophenotypes and the identification of novel circulating B-cell subsets defined by the expression of CD20, CD5, and CD11c antigens. Morphologic evaluation of 14CD11c+CD5+ B-cell leukemias showed that they generally had larger cellular diameters (14 to 21 microns) and lower nuclear:cytoplasm ratios than typical small lymphocyte CLL. These cases did not exhibit the well-defined nucleoli characteristic of prolymphocytic leukemia (PLL). The presenting clinical features of CD11c+CD5+ B-cell leukemias were most consistent with CLL or PLL, and none of the evaluated cases had pancytopenia, splenomegaly, and cytoplasmic villi characteristic of HCL. Examination of normal peripheral blood (n = 6) by three-color flow cytometry identified four novel B-cell subsets with the following immunophenotypes (mean percent of total CD20+ B cells +/- SE): CD20+CD5+CD11c+ (8.0 +/- 1.6); CD20+CD5-CD11c+ (12.0 +/- 2.0); CD20+CD5+CD11c- (35.0 +/- 4.9); and CD20+CD5-CD11c- (44.0 +/- 5.0). Our findings suggest that CD11c+CD5+ B-cell leukemias with atypical morphologic features represent forms of CLL or PLL rather than HCL. In addition, we have identified novel subsets of circulating B cells defined by patterns of CD20, CD5, and CD11c expression that correspond to the immunophenotypes of chronic B-cell leukemias.

Adult↗

PubMed1989

Immunophenotypic and antigen receptor gene rearrangement analysis in T cell neoplasia.

The author reviews the immunophenotypic profiles displayed by the major clinicopathologic categories of T cell neoplasia, the immunophenotypic criteria useful in the immunodiagnosis of T cell neoplasia, and the contributions made by antigen receptor gene rearrangement analysis to the understanding of T cell neoplasia. Neoplasms belonging to distinct clinicopathologic categories of T cell neoplasia often exhibit characteristic immunophenotypic profiles. Approximately 80% of lymphoblastic lymphomas and 20% of acute lymphoblastic leukemias express phenotypes consistent with prethymic and intrathymic stages of T cell differentiation, including intranuclear terminal deoxynucleotidyl transferase. Cutaneous T cell lymphomas of mycosis fungoides type usually express pan-T cell antigens CD2, CD5, and CD3, often lack the pan-T cell antigen CD7, and usually express the mature, peripheral helper subset phenotype, CD4+ CD8-. Cutaneous T cell lymphomas of nonmycosis fungoides type and peripheral T cell lymphomas often lack one or more pan-T cell antigens and, in addition, occasionally express the anomalous CD4+ CD8+ or CD4- CD8- phenotypes. T gamma-lymphoproliferative disease is divisable into two broad categories: those cases that are CD3 antigen positive and exhibit clonal T cell receptor beta chain (TCR-beta) gene rearrangements and those cases that are CD3 antigen negative and exhibit the TCR-beta gene germline configuration. Human T cell lymphotropic virus-I (HTLV-I) associated Japanese, Carribean, and sporadic adult T cell leukemia/lymphomas usually express pan-T cell antigens, the CD4+ CD8- phenotype, and various T cell-associated activation antigens, including the interleukin-2 receptor (CD25). Immunophenotypic criteria useful in the immunodiagnosis of T cell neoplasia include, in increasing order of utility, T cell predominance, T cell subset antigen restriction, anomalous T cell subset antigen expression, and deletion of one or more pan-T cell antigens. Only in exceptional circumstances do normal, non-neoplastic T cell populations express the CD4- CD8- or the CD4+ CD8+ phenotype and/or lack one or more pan-T cell antigens. T cell receptor beta chain gene rearrangement analysis represents an accurate, objective, and sensitive molecular genetic marker of T cell lineage and clonality that allows discrimination among non-T cell, polyclonal T cell and monoclonal T cell populations. Non-T cells exhibit the TCR-beta gene germline configuration.(ABSTRACT TRUNCATED AT 400 WORDS)

Cell Differentiation↗

PubMed1989

Prognostic importance of the pre-B-cell immunophenotype and other presenting features in B-lineage childhood acute lymphoblastic leukemia: a Pediatric Oncology Group study.

We report the prognostic significance of the pre-B-cell immunophenotype and other presenting features, including blast cell karyotype, in a randomized clinical trial conducted from 1981 to 1986 for children with early pre-B (n = 685) or pre-B (n = 222) acute lymphoblastic leukemia (ALL). Patients greater than or equal to 1 year and less than or equal to 21 years of age who attained complete remission were stratified by conventional risk criteria and immunophenotype and then randomized to receive continuation therapy with either of two regimens of intensive chemotherapy, designated S (standard) and SAM (standard plus intermediate-dose methotrexate, 1 g/m2 every 8 weeks). The proportions of subjects achieving complete remission in the two phenotypically defined subgroups were identical, 96%. At a median follow-up time of 42 months, the overall probability of 4-year event-free survival (+/- SE) was 63% +/- 2% (pre-B = 51% +/- 5% and early pre-B = 66% +/- 3%). Children with pre-B ALL had significantly shorter durations of continuous complete remission (P = .0004); this association included both bone marrow and CNS remissions (P = .0004 and P = .02, respectively). In a univariate Cox regression analysis of potentially important prognostic factors, the pre-B immunophenotype was significantly related to a poorer outcome, as were other recognized biologic and clinical features (eg, pseudodiploidy, older age, male sex, black race, and a higher WBC). It retained its prognostic strength in a multivariate model based on age, WBC, ploidy, and sex. The risk of failure at any point in the clinical course of a child with the pre-B immunophenotype was 1.8 times as great as that in a patient lacking this feature but otherwise having an equivalent risk status. It should be stressed that the predictive value of any of the significant characteristics identified in this study could diminish in the context of another, more effective treatment program. Nevertheless, our major conclusion, that children with pre-B ALL fare worse than those with early pre-B disease in a contemporary clinical trial has implications for stratified randomization of patients and the design of risk-specific treatment protocols.

Analysis of Variance↗

PubMed1989

Rapid diagnosis of leukaemia: a three-stage immunophenotyping technique.

We have developed a three-stage avidin-biotin alkaline phosphatase system (ABAP) for immunophenotyping acute and chronic leukaemias. Air-dried blood and bone marrow films or cytospins were fixed in acetone, methanol-acetone, or methanol-acetone-formalin. A primary monoclonal antibody layer was employed with a corresponding biotinylated anti-species second layer. The third stage was avidin-conjugated alkaline phosphatase and visualisation was achieved with Fast Red TR as diazo salt. This was compared using a limited range of primary antibodies with the alkaline phosphatase anti-alkaline phosphatase (APAAP) method, routinely performed as a five-stage technique. The ABAP technique provided a 2.5 h, reliable, three-stage immunophenotyping procedure without the need for amplification steps. A wider range of first layer monoclonals of any species could be utilised by using the appropriate secondary antibody. Preservation of cell morphology was very good. Smaller amounts of expensive monoclonal antibodies could be used compared to the quantity required with other slide techniques. This approach to immunophenotyping can consequently be employed by any general haematology laboratory to provide a sensitive and inexpensive service. The technique has been in routine diagnostic use for the last 2 years, and consistently good results have so far been achieved in a national external immunophenotyping quality assurance scheme.

Acute Disease↗

PubMed1989

Organization and operation of a flow cytometric immunophenotyping laboratory.

Technical advances in the field of flow cytometry have made it feasible for many academic and private hospital laboratories to purchase relatively inexpensive "user friendly" flow cytometers that do not require dedicated flow cytometer operators, special rooms, or a significant amount of laboratory space. Because the financial and physical constraints in operating a flow cytometer have been substantially reduced, many pathologists may now be considering such a purchase. By chronicling the clinical activities of a single flow cytometric immunophenotyping laboratory, this report will answer a variety of questions that may be asked by pathologists regarding both the utility of flow cytometers in clinical diagnosis and the mechanics of operating an immunophenotyping laboratory. The types of tissues that can be evaluated by flow cytometry will be detailed, and we will summarize the number and type of flow cytometric clinical studies performed in our laboratory since its development in 1983. Practical aspects of laboratory operation including technical staff requirements, specimen handling and processing procedures, monoclonal antibody selection, and quality control procedures will be presented. In addition, a comprehensive review of flow cytometric immunophenotyping studies as applied to the diagnosis of leukemias, lymphomas, and immunodeficiency disorders will be presented along with case examples that illustrate our approach to the interpretation of immunophenotyping results.

Adult↗

PubMed1987

Immunophenotypic criteria for the diagnosis of non-Hodgkin's lymphoma.

This study examines the immunohistologic profiles of a large series of histologically proven benign and malignant lymphoproliferative processes in order to define immunophenotypic criteria useful in the diagnosis of non-Hodgkin's lymphoma. Using a method of analysis relying solely on immunoarchitectural features of a given case, the authors were able to define immunologic criteria capable of differentiating benign from malignant lymphoid processes independent from conventional morphologic analysis. In general, these criteria involved identification of abnormal expression or loss of antigens in B- and T-lineage populations. Among B-lineage populations the following features were associated with malignant histology: 1) light-chain-restricted B lineage, 2) light chain -B lineage, 3) Leu-1+ B lineage, 4) L60+ B lineage, 5) 41H+, Ki-67+ B lineage, 6) loss of pan-B antigens, and 7) LFA-1-B lineage. Among T-cell populations outside the thymus, phenotypes associated with malignancy included 1) loss of pan-T antigens (including loss of the beta chain of the T-cell antigen receptor), 2) coexpression or loss of T-subset antigens, 3) Leu-6+ T-lineage, and 4) MB-1+ T lineage. Application of these criteria to a series of nearly 500 cases of lymphoma indicated that over 90% of B-lineage and about 80% of T-lineage neoplasms manifested immunophenotypic abnormalities that could distinguish them from benign, reactive lymphoid processes. It is concluded that immunophenotypic analysis of lymphoproliferative lesions is sufficiently sensitive and specific to confirm the histologic diagnosis of lymphoma in the vast majority of cases seen in clinical practice. Furthermore, in difficult cases or those with limited material or poor histology, immunophenotypic analysis may be the only means of making a definitive diagnosis.

Antibodies, Monoclonal↗

PubMed2000

Diagnosis of unexpected acute myeloid leukemia and chronic lymphocytic leukemia: a case report demonstrating the perils of restricted panels in flow cytometric immunophenotyping.

We report on the flow cytometric identification of concomitant acute myeloid leukemia and chronic lymphocytic leukemia in cytology specimens submitted with minimal clinical information. A 64-year-old man presented with fever and progressive dyspnea on exertion. Chest X-ray and computed tomography scan showed a left upper lobe pulmonary mass. Pulmonary capillary pullback specimens were collected to determine infectious verses neoplastic etiology. The pulmonary capillary pullback specimens showed atypical mononuclear cells with enlarged, slightly irregular nuclei; visible nucleoli; and basophilic cytoplasm. Flow cytometric analysis of the specimen for lymphoma was requested. Flow cytometric immunophenotypic studies showed that 78% of the cells were CD34 positive, CD45 dim positive and CD11c positive, consistent with acute myeloid leukemia. About 0. 75% of the cells expressed CD5 as well as dim CD20 and were monoclonal for kappa light chains: consistent with chronic lymphocytic leukemia/small lymphocytic lymphoma. At this time the clinician communicated a history of myelodysplastic syndrome of refractory anemia subtype. Peripheral blood was obtained for further immunophenotyping and the patient was immediately treated for his acute myeloid leukemia. This case demonstrates that a diagnostic antibody panel should allow evaluation of all cell types as per the U.S./Canadian consensus recommendations on the immunophenotypic analysis of hematologic neoplasia by flow cytometry (Stewart et al.: Cytometry 30:231-235, 1997). Published 2000 Wiley-Liss, Inc.

Acute Disease↗

PubMed1999

Fine-needle aspiration with flow cytometric immunophenotyping for primary diagnosis of intra-abdominal lymphomas.

Cytomorphology in conjunction with immunophenotypic characterization is becoming increasingly used for the primary diagnosis of non-Hodgkin's lymphomas (NHL). This combination is especially advantageous for the diagnosis of intra-abdominal and intrathoracic lymphomas, since unlike superficial lesions, open biopsy of deep-seated tissues is more invasive and more costly, and is associated with a higher risk. We report the cytologic and immunophenotypic features of intra-abdominal NHL obtained by fine-needle aspiration (FNA). Twenty-two cases of intra-abdominal lesions obtained by image-guided FNA where flow cytometry was also performed were reviewed. Of the 22 studied cases, 7 were classified as large-cell lymphoma, 5 as follicular center-cell lymphoma, 2 as small noncleaved-cell lymphoma, 2 as lymphoplasmacytoid lymphoma, one as small lymphocytic lymphoma, and one as marginal-zone lymphoma. In the remaining 4 cases where the immunophenotypic pattern was not definitive, the cytomorphologic features were of small cleaved cells in 3 cases and of mixed small cleaved and large cells in one case. We successfully classified 9 of the 10 patients on whom histologic confirmation was obtained. The successful primary classification of most intra-abdominal non-Hodgkin's lymphomas can be done with a combination of cytology and flow cytometry, and this can be the initial approach in patients with deep-seated lesions. Diagn. Cytopathol. 1999;21:98-104.

Abdominal Neoplasms↗