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PubMed1999

Blast crisis of chronic myelogenous leukemia exhibiting immunophenotypic features of a myeloid/natural killer cell precursor.

We report a patient with Philadelphia chromosome (Ph1)-positive chronic myelogenous leukemia (CML) which transformed into blast crisis bearing the immunophenotypic features similar to those of the myeloid/natural killer (NK) cell precursor leukemia we proposed previously. Using a CD45 blast gating method, the myeloperoxidase-negative blasts were positive for CD7, CD13, CD33, CD34, CD56, and HLA-DR, but no other lymphoid antigens. Southern blot analysis showed germ line T cell receptor beta and delta genes and immunoglobulin heavy and light chain genes. Although NK cell blastic transformation with Ph1 positive CML has been reported in a single patient, this is, to our knowledge, the first report of CML blast crisis of myeloid/NK cell precursor origin.

Aged↗
PubMed1994

An expert system for the interpretation of flow cytometric immunophenotyping data.

The development of high-grade non-Hodgkin's lymphomas in HIV-positive patients and patients with acquired immune deficiency syndrome (AIDS) is a well known phenomenon. The proper classification of these neoplasms often requires a multiparameter approach, including the interpretation of a large panel of immunologic markers analyzed by flow cytometry. The availability of individuals with the required expertise to properly interpret these marker studies is limited. For this reason, we have designed an expert system to automate the analysis of immunophenotyping panels in both HIV-related and non-HIV-related hematopoietic neoplasms. The expert system, which we call "Professor Fidelio", runs on IBM-compatible computers under Windows 3.0. The system is designed to accept any number of markers studied from a repertoire of 35 markers. Professor Fidelio functions on the basis of heuristic classification of defined diagnostic patterns. Nine specific patterns (Stem Cell, Myeloid and/or Monocytic, Erythroid, Megakaryocytic, Immature B-cell, Immature T-cell, Mature B-cell, Mature T-cell, and Plasma cell) and one "non-specific" pattern have been agreed upon. Fidelio's knowledge base contains the definitions of each of these patterns and the heuristics for excluding patterns when an incomplete panel of markers is performed. The inference engine interprets the findings (including the age of the patient) and reports the patterns which are matched, the differential diagnosis, the suggested diagnosis from the list of differentials if the marker studies are specific, and recommendations for additional tests which may be valuable in establishing the diagnosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Acquired Immunodeficiency Syndrome↗
PubMed1998

Evaluation of monoclonal antibody reagents from three different manufacturers using flow cytometric two-color immunophenotyping.

We evaluated a flow cytometric (FCM) two-color immunophenotyping of CD3+/CD4+ T-helper and CD3+/CD8+ T-suppressor lymphocytes in whole blood samples from HIV-infected individuals using monoclonal antibody reagents from three different manufacturers. Lymphocytes were firstly determined using CD45/CD14 in association with a forward scatter/side scatter gating strategy. CD3+/CD4+ and CD3+/CD8+ were then determined and compared. Reagents from all manufacturers showed good separation of lymphocytes, monocytes and granulocytes with high purity and recovery. There was a good correlation of the percentage of CD3+/CD4+ and CD3+/CD8+ lymphocytes amongst each of the manufacturer's reagents, but the fluorescent intensities of positive cells were not the same. This difference can result in poor discrimination of positive and negative non CD3 cells leading to erroneous results.

Antibodies, Monoclonal↗
PubMed1999

Leukaemia-associated immunophenotypes (LAIP) are observed in 90% of adult and childhood acute lymphoblastic leukaemia: detection in remission marrow predicts outcome.

Analysis of differentiation antigens on leukaemic blasts is routinely done for diagnostic purposes, i.e. determination of stage of differentiation and lineage assignment. Acute lymphoblastic leukaemias are also frequently characterized by a leukaemia-associated immunophenotype (LAIP), either the coexpression of differentiation antigens physiologically restricted to other stages of differentiation (asynchronous LAIP) or cell lineages (aberrant LAIP). We defined LAIP in 241 consecutive unselected B-lineage (n = 193) and T-lineage (n = 48) ALL by three-colour flow cytometry using directly conjugated monoclonal antibodies. The incidence of LAIP was found to be 91.7%. In 63% of patients two to six leukaemia-associated expression patterns were detected. In order to study the specificity of LAIP in a therapy-relevant setting, remission bone marrow samples from patients with B-lineage ALL were analysed for the expression of T-lineage-associated phenotypes on the normal bone marrow cells and vice versa. The frequency of all T-lineage LAIP+ cells and all aberrant B-lineage LAIP+ cells was <1% in regenerating bone marrow samples at different timepoints. The incidence and clinical significance of LAIP+ cells was studied in 196 remission marrows of 70 ALL patients (55 remaining in CCR, 14 with bone marrow relapse, one with isolated CNS relapse). The presence of >1% LAIP+ at two consecutive timepoints predicted 5/8 bone marrow relapses in B-lineage ALL. The occurrence of LAIP+ cells >1% in T-lineage ALL after induction therapy predicted relapse in 7/7 cases. In conclusion, flow cytometric detection of LAIP+ cells appears to be a powerful tool for the prediction of outcome in ALL.

Adolescent↗
PubMed1999

Belgian consensus recommendations for flow cytometric immunophenotyping. The Belgian Association for Cytometry/Belgische Vereniging voor Cytometrie/Association Belge de Cytométrie.

This paper summarises the guidelines and recommendations that were generated during a number of discussion forums attended by the majority of Belgian cytometry laboratory professionals. These forums focused on the rational and optimal use of flow cytometric evaluations in the clinical laboratory setting. The aim was to improve the coherence of the testing panels and the quality of the results and--as such--the clinical diagnostic information. It was also the aim to provide the Belgian prescribing physician and interested laymen with an updated overview of the flow cytometric possibilities. Emphasis is placed on immunophenotyping of haematological malignancies, hematopoietic progenitor cell counting and follow-up of the viral infection caused by the human immunodeficiency virus.

Acquired Immunodeficiency Syndrome↗
PubMed1999

Multicenter study of reference stabilized human blood for lymphocyte immunophenotyping quality control in flow cytometry. GEIL.

The development of automated methods and normative rules, as well as the dependence of some therapeutic approaches on lymphocyte subsets counts, has led to the appearance of calibration reagents. Such reagents are expected to perform equally well in very different settings. We developed a multicenter trial to evaluate the performance of a new quality control reagent, i.e., stabilized blood to be used in immunophenotyping laboratories. Aliquots of the same batch of stabilized blood were shipped to 45 French and Belgian laboratories on a Monday and had to be tested for percentages and absolute counts of at least CD3-, CD4-, and CD8-positive lymphocytes on 4 consecutive days. The percentages and absolute counts obtained on each assay were recorded, as well as the type of lysis used, the trademark of reagents, and the brand of flow cytometer. The mean values collected did not differ significantly from those expected by the manufacturer. Absolute counts generated through one-step techniques displayed lower CVs. This new reagent therefore appears to be a robust product, liable to yield consistent results in the array of different conditions represented by routine laboratories, and it could be useful for quality control procedures.

Flow Cytometry↗
PubMed1999

[Immunophenotyping of acute lymphoblastic leukemia in Mexican children].

PURPOSE: To analyse the immunophenotype of leukaemic cells in a group of children diagnosed of lymphoblastic leukaemia in order to assess the frequency of the different immunologic subtypes. PATIENTS AND METHODS: In the period comprised between APR 1987 and MAR 1995, 402 Mexican children were studied in a prospective way. Conventional immunological markers were used, either associated to or specific for B, T, myelo-monocytic or megakaryocytic-platelet cell populations. RESULTS: Five major immunologic subtypes were disclosed, showing a series of specific surface markers: null-ALL, 5%; early pre-B, 7.5%; common, 74.6%; B-cell, 3.5%, and T-cell, 9.4%. A net predominance of B-cell precursor CD10- ALL was found in children under one year of age, and of CD10+ B-cells beyond that age. Although there was only slight predominance of male sex, the prevalence of B and TALL in males was not confirmed. CONCLUSIONS: These results show that the incidence of the different immunologic subtypes of lymphoblastic leukaemias and their distribution according to age and sex are closely similar to those reported among Caucasians in other parts of the world.

Adolescent↗
PubMed1999

Immunophenotyping by flow cytometry of fine needle aspirates in the diagnosis of lymphoproliferative disorders: A retrospective study.

In order to determine the value of flow cytometric (FCM) immunophenotyping of fine-needle aspirates (FNA) in the diagnosis and classification of lymphoproliferative diseases, 61 tissue samples were studied and compared with the cytologic/histological results. In vivo and ex vivo FNA biopsy yielded the material for FCM, which comprised an extensive number of lymphoid cell markers. In all but three cases sufficient cells were collected. Overall, malignancy was diagnosed in 33 cases from a total of 47 (70.2%), and in the remaining cases malignancy was not detected. Eleven cases were correctly diagnosed as reactive processes (11/11). There were no false positive cases of malignancy, as diagnosed by FCM-FNA. The best accuracy was achieved in the low-grade B-cell lymphomas and lymphoblastic lymphoma/leukemia. We conclude that in a significant number of cases, FCM-FNA permits the separation between lymphoid malignancies and reactive processes without false positive results. It was found to be particularly useful in the differential diagnosis of mantle-cell and small-lymphocytic lymphoma and in the identification of lymphoblastic lymphoma/leukemia.

Antibodies, Monoclonal↗
PubMed1999

Histopathology of typhoid enteritis: morphologic and immunophenotypic findings.

Enteric fever is a systemic illness caused by Salmonella infection, with S. typhi, S. paratyphi, and S. enteritidis being the most common serotypes. Humans are the only reservoir for S. typhi, and its predilection for the ileum is due to the fact that organisms enter the body by translocation across specialized Peyer's patch epithelium and then proliferate in the mucosal macrophages. The lesions in bowel and mesenteric lymph nodes are distinctive and mimic Kikuchi-Fujimoto disease and Rosai-Dorfman disease as well as infections caused by some non-salmonella bacteria. The four cases presented in this report, two culture-confirmed, all exhibited ileal mucosal hypertrophy caused by a neutrophil-poor monocyte/macrophage-rich hyperplasia. Though diffuse areas were present, much of the lesional proliferation was nodular, representing macrophage infiltration and colonization by the monocytes and macrophages. Immunophenotypic studies, which showed a CD68+, lysozyme+, UCHL-1+, OPD4-, CD4-, s100- profile, were helpful in distinguishing these lesions from other processes, including Kikuchi-Fujimoto disease and Rosai-Dorfman disease. Although rare in developed countries, enteric fever should be considered in any patient with recent travel to endemic areas and in the context of illness thought to be related to contaminated foods.

Adult↗
PubMed1999

[Stromal tumors of the small intestine (GIST). Prognostic differences based on clinical, morphological and immunophenotypic features].

Gastro Intestinal Stromal Tumors (GIST) are rare malignant non-epithelial tumors arising from gastro-intestinal tract. They represent the 0.2% of all malignancy of this site. Three cases, treated in our Department, are an example of the anatomo-clinical features of these tumors. In two cases the clinical features on admission were similar to those of acute abdominal pain; in the third one a palpable mass rapidly growing was observed. All three patients had surgical treatment but survival were quite different. The size of tumor mass in the first two cases was 5 cm; one patient was alive after ten years without relapse, the second one died five years after with liver metastasis. The third one, with a mass larger more than ten centimeters, developed 8 months after surgical resection a diffuse local relapse of the disease. The microscopic pattern and the lack of epithelial markers allowed to classify them as GIST. All cases were tested with S100 protein, smooth muscle actine, and CD-34 antigen. Two cases had both smooth muscle and nervous markers, the third resembled a malignant schwannoma. Prognosis was correlated with the size of tumor at the time of surgery and immunophenotype pattern. The tumors with high expression of nervous antigens had a worse prognosis.

Antigens, CD34↗
PubMedDate not supplied

Immunophenotypical analysis and immunobiology of childhood brain tumors.

Cancer associated markers (CAMs) are the biochemical and immunological counterparts of the morphology of neoplasms. The expression of an immunocytochemically defined CAM is related to the tissue of origin and is not a random event. During the past two decades, the use of MoABs against oncofetal, neoplasm associated, cell lineage specific, endothelial, and cell proliferation related antigens in the diagnosis and biological assessment of prognosis in neoplastic disease gained increased importance. A sensitive direct correlation exists between the expression of certain molecules and the development of an invasive, highly malignant immunophenotype (IP) of neoplastic cells, allowing for the occurrence of angiogenesis and metastasis. Our systematic and detailed cellular IP analyses of 82 childhood brain tumors [34 medulloblastomas (MEDs)/primitive neuroectodermal tumors (PNETs), 42 astrocytomas (ASTRs), 5 choroid plexus papillomas (CPPs) and 1 choroid plexus carcinoma (CPC)], was conducted using over 55 MoABs. An indirect, four-step, enzyme linked [alkaline phosphatase (AP) and peroxidase (PO)], biotin-streptavidin based, antigen detection technique was employed. (ABSTRACT TRUNCATED)

Brain Neoplasms↗
PubMed2000

Childhood non-Hodgkin's lymphoma: an immunophenotypic analysis.

OBJECTIVE: To observe the frequency of histological subtypes of childhood non-Hodgkin's lymphoma and its immunohistochemical profile. SETTING: All cases of non-Hodgkin's lymphoma diagnosed in children (< 15 years) in the section of histopathology at the Aga Khan University Hospital Karachi during the period of three years. METHODS: These consecutive cases were evaluated on H&E stained sections and then immunohistochemistry analysis of these tumors was performed by employing Peroxidase Anti-Peroxidase (PAP) technique. RESULTS: The present series included 61 cases of non-Hodgkin's lymphoma. NHL was more common in males as compared to females with male to female ratio of 5.8:1. High grade NHL comprised 87% of childhood lymphoma. The mode of presentation in majority of NHL (57%) was extranodal. Burkitt's lymphoma (33%) was the most prevalent histological subtypes, followed by lymphoblastic (28%), diffuse large cell (15%), diffuse mix small and large cell (13%), small non cleaved Non-Burkitt's (7%) and immunoblastic (4%). Immunophenotypic analysis of the childhood Non-Hodgkin's lymphoma revealed that 67% of the Non-Hodgkin's lymphoma are B-cell type while 33%, are those of T-cell lineage. CONCLUSION: NHL was more common in males. Majority of NHL in children were high grade tumors. Burkitt's lymphoma was the most frequent histological subtype. T-cell NHL comprised a significant portion of childhood lymphomas.

Adolescent↗
PubMedDate not supplied

[The immunophenotypic and cytogenetic characteristics of the blast cells in subvariants of acute myeloid leukemia].

The results of study of morpho-cytochemical peculiarities, antigenic profile and peripheric blood and/or bone marrow blast cell karyotype of 21 patients with acute myeloid leukemia are presented. Hemopoietic cell immunophenotyping was carried out with the use of cytofluorometer FACScan and chromosome cytogenetic analysis with the use of analyzer "Metascan". It has been shown that in the AML M1 blast cell plasmatic membrane carries pan-myeloid CD33 and CD13 antigens, the last having high density of expression, and the CD38 antigen, which is a myeloid cell-precursor marker. In these patients tetraploidy, being the testimony of karyotype change, has been ascertained. It has been found out that the AML M2 blasts, except pan-myeloid antigens, express the cell proliferation CD71 marker. Blast cell karyotype peculiarities typical for this leukemia sub-variant have been revealed. In patients with the AML M4 in 3 of 6 cases an anomalous karyotype has been found. It has been also shown that the CD14 antigen, and rather its percentage in blast total population, is the differential-diagnostic criterion for the AML M4 and M5.

Acute Disease↗
PubMed1995

Immunophenotyping of acute leukemia by flow cytometry: closing the black holes of morphology.

The heterogeneity of disease conditions in acute leukemia (AL) requires the most discriminative procedures for a differential diagnostic classification. Selected cases of AL, representative of lineage fidelity, multilineage phenotype and phenotypic switch, are reviewed to document that the diversity of the leukemic cells (lineage involved, maturation stage, aberrant markers), although not always characterizable by morphological differences, is reflected by a variety of immunophenotypes which may be efficiently characterized by flow cytometry to obtain a precise diagnosis.

Acute Disease↗
PubMed2000

Immunophenotyping of hematopoietic neoplasms.

Advances in the staging and treatment of hematopoietic neoplasms have necessitated a high degree of accuracy in the diagnosis and classification of these tumors. A greater degree of diagnostic precision has resulted from recent advances in immunophenotyping and genotyping of hematopoietic neoplasms. This review discusses several new immunohistochemical reagents, many of which are derived from results of molecular studies.

Antibodies↗
PubMedDate not supplied

[Classification of acute leukemias according to the first latin-american consensus conference for the immunophenotyping of leukemias].

OBJECTIVE: To evaluate the recommendations of the First Latinamerican Consensus Conference for the Immunophenotyping of Acute Leukemia in untreated patients with de novo disease immunologically classified employing flow cytometry and an extended panel of monoclonal antibodies. MATERIAL AND METHODS: In that conference it was recommended the use of the following antibodies: cytoplasmic CD79a (cCD79a) and CD19 to define B-progenitor acute lymphoblastic leukemia (B-ALL); cCD3 and CD7 for T-cell ALL (T-ALL), and CD13, CD33 and myeloperoxidase (cMPO) for acute myeloblastic leukemia (AML). We analyzed the expression of these cellular antigens in 91 non-consecutive patients classified with the extended panel as: B-ALL 28 cases; T-ALL 7; B-T-ALL 2; AML 47; and mixed-lineage acute leukemia 7 cases. RESULTS: All 28 B-ALL cases were positive with each of the two recommended antibodies cCD79a and CD19, whereas in 24 AML cases (the expression of cCD79a was not assayed in 23 cases) and in 7 T-ALL patients both antigens were absent. cCD3 and CD7 antigens were identified in 71% and 100% of T-ALL, respectively. CD7 antigen was not detected in any of the 28 patients with B-ALL but it was expressed in 6 of 47 AML cases, while none of 75 B-ALL and AML cases were positive to cCD3. Forty nine percent of AML were positive for the three recommended markers: cMPO, CD13 and CD33, and 51% of AML cases reacted with one or two of these three monoclonal antibodies. Six out of 28 cases of B-ALL had aberrant expression of myeloid antigen (CD33 in 3 cases and CD13 in 3 cases). CONCLUSIONS: There was no difference in the definition of AL lineage between employing the extended antibody panel and that recommended by the Latinamerican consensus.

Acute Disease↗
PubMed1998

[Investigation of the clinicopathologic features, immunophenotype and Epstein-Barr virus infection of the upper respiratory tract lymphomas in patients].

OBJECTIVE: To study the clinicopathologic features, immunophenotype and Epstein-Barr virus (EBV) infection of the upper respiratory tract lymphomas in patients from north China. METHODS: 112 cases of upper respiratory tract lymphomas in patients from north China were investigated. HE stain for histologic features. Immunohistochemistry studies were performed using monoclonal antibodies, UCHL-1 for T-lymphocyte, L26 and 4KB5 for B lymphocyte, CS1-4 for latent membrane protein-1 (LMP-1). In situ hybridization for EBV encoded small nuclear RNA (EBER 1/2, Y017, DAKO). RESULTS: 77/112 cases (68.75%) were T-cell lymphomas, 35 cases (31.25%) were B cell lymphomas. EBV-EBER 1/2 in situ hybridization positive in 54/101 (53.46%) cases, including 48 T-cell lymphoma cases and 6 B-cell lymphoma cases. CONCLUSION: Most upper respiratory tract lymphoma cases in patients from north China are T-cell lymphomas and are strongly associated with EBV.

Adolescent↗
PubMed2001

Clinicopathologic, immunophenotypic, and molecular characterization of primary cutaneous follicular B-cell lymphoma.

OBJECTIVE: To determine the clinicopathologic, immunophenotypic, and molecular characteristics of primary follicular cutaneous B-cell lymphoma (CBCL) as defined by the revised European-American lymphoma classification. DESIGN: A retrospective survey of the medical records, an immunohistochemical study of archival biopsy specimens. and molecular studies of preserved DNA of all patients with follicle center lymphoma-follicular (FCL-F) primary CBCL from 1987 to 1997. SETTING: A single-center outpatient specialty clinic at an academic medical center. PATIENTS: Twenty-one patients (68% of all new primary CBCL cases), including 14 men and 7 women (age range, 33-88 years; mean, 55 years). RESULTS: The head and neck region was the most frequent primary site. Following treatment, recurrences were relatively frequent, but the overall mortality rate during 1.0 to 11.3 years (mean, 6.3 years) of follow-up was 4.8%. Immunohistochemical analysis for B- and T-cell lineages was helpful in enhancing the folliclelike structures. CD10, bcl-2, and CD43 were expressed by the neoplastic cells in 9 (47%) of 19 cases, 4 (21%) of 19 cases, and 2 (13%) of 16 cases, respectively. Immunohistochemical detection of cytoplasmic immunoglobulin light chains, using steaming in EDTA as the antigen-retrieval technique, was successful in 12 (71%) of 17 cases. The Ig heavy-chain gene rearrangements, using the Southern blot technique, detected clonality in 17 (94%) of 18 cases. The bcl-2 gene rearrangements were detected in only 2 (13%) of 15 of the primary cutaneous FCL-F cases, compared with 9 (75%) of 12 of the primary nodal FCL-F cases (P =.002). CONCLUSIONS: Primary cutaneous FCL-F is a relatively common subtype of CBCL, with a relatively indolent course. It has many features in common with primary nodal FCL-F, except for low rates of bcl-2 expression and bcl-2 gene rearrangements.

Adult↗