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At least 19 recordsLinked to original sources
PubMed2004

Applications of flow cytometry and immunohistochemistry to diagnostic hematopathology.

OBJECTIVE: Diagnostic hematopathology depends on the applications of flow cytometric immunophenotyping and immunohistochemical immunophenotyping combined with the cytomorphology and histologic features of each case. Select cases may require additional ancillary cytogenetic and molecular studies for diagnosis. The purpose of this review is to focus on the applications of flow cytometric and immunohistochemical immunophenotyping of paraffin-embedded tissue to diagnostic hematopathology. Advantages and disadvantages of these techniques are examined. DATA SOURCES: The literature is extensively reviewed (PubMed 1985-2003) with an emphasis on the most recent applications and those that are most useful clinically, both diagnostically and prognostically. Study Selection.-Studies were selected based on statistically significant results in large studies with reported adequate clinical follow-up. DATA EXTRACTION: The methodology was reviewed in the selected studies to ensure reliable comparison of reported data. DATA SYNTHESIS: Flow cytometric immunophenotyping offers the sensitive detection of antigens for which antibodies may not be available for paraffin immunohistochemical immunophenotyping. However, paraffin immunohistochemical immunophenotyping offers preservation of architecture and evaluation of expression of some proteins, which may not be available by flow cytometric immunophenotyping. These techniques should be used as complimentary tools in diagnostic hematopathology. CONCLUSIONS: There are extensive applications of flow cytometric and immunohistochemical immunophenotyping to diagnostic hematopathology. As cytogenetic and molecular findings evolve in diagnostic hematopathology, there may be additional applications of flow cytometric and immunohistochemical immunophenotyping to this field of pathology.

Flow Cytometry↗
PubMedDate not supplied

Controversies on Hodgkin's disease and anaplastic large cell lymphoma. Hematopathology Study Group of the Società Italiana di Anatomia Patologica.

Just one year ago the Italian Society of Pathology (S.I.A.P.) created a Study Group which included members of the most active Italian hematopathology teams. Prof. Pasquale Calapso was asked to chair the Group and Prof. Stefano Pileri to take care of secretarial duties. The aim of the Group is to spread hematopathologic knowledge among young pathologists and to promote activities that can contribute to updating Italian pathologists on topics of both speculative and diagnostic interest. The first Workshop of the S.I.A.P. Hematopathology Group was held at the Palazzo dei Congressi in Bologna, November 20, 1993. About 150 pathologists from all over Italy took part in the meeting, which consisted of two sections devoted to: a) discussion of the boundaries between Hodgkin's disease and non-Hodgkin's lymphomas, and b) a case seminar illustrating the impact of immunohistochemistry in the diagnosis of bone-marrow biopsy. The first section included 5 presentations and a Round Table chaired by Prof. Luciano Fiore-Donati. Below, the contributors to this section summarize the content of their presentations, which were aimed at answering specific questions the Organizers had put to them.

Hodgkin Disease↗
PubMed1994

Maude Abbott Lecture. Hematopathology: a crescendo of scholarly activity.

This lecture traces the evolution of hematopathology from the time of Thomas Hodgkin to the modern era of hybridoma antibodies, immunophenotyping, and molecular biology. It highlights the early concepts of Ludwig Aschoff and his school based on the observations of Metchnikoff, Ehrlich, and Maximow and the changes in concepts and terminology that have occurred in the ensuing 80 yr. The ongoing controversy on terminology and classifications of the malignant lymphomas from their first descriptions by Rudolf Virchow, Hans Kundrat, and Theodore Billroth is reviewed with reference to the development of the Working Formulation of Non-Hodgkin's Lymphomas for Clinical Usage and other classifications that are currently in vogue. Special attention is paid to the remarkable insight showed by Dorothy Reed in her morphologic descriptions of Hodgkin's disease, her thoughtful discussion on the nature of this disorder, and the conformation of her proposals inherent in the subsequent classification of Lukes and Butler. Acknowledgement is given to the remarkable discovery of Köhler and Milstein resulting in the advent of monoclonal antibodies that have revolutionized the field of hematopathology; to the many immunopathologists who have been responsible for the production of more than 1100 monoclonal antibodies comprising 78 clusters of differentiation; and to many of the leading molecular biologists who have (in the words of Berard) "helped to transform hematopathology from a difficult morphologic exercise into a functionally oriented biologic science."

Hematology↗
PubMed2005

The hematopathological basis for studying effects of the demethylating agent 5-aza-2'-deoxycytidine (decitabine) in myelodysplasia.

The myelodysplastic syndromes have, since their first recognition decades ago, been considered notoriously difficult with regard to their proper classification, determination of prognosis, and optimal treatment. With the advent of the French-American-British (FAB) classification, now aided but not superseded by the World Health Organization classification, distinct biological entities have been delineated, which in turn are very useful for stratification to different, established and experimental treatment modalities. However, precise subclassification of different types of myelodysplastic syndrome (MDS) is only possible with hematopathological studies based on the analysis of peripheral blood, bone marrow smear, and bone marrow biopsy, backed by appropriate clinical information. Bone marrow cytogenetics are also essential for any risk stratification since they still provide the second most powerful prognostic parameter after bone marrow blast enumeration. This paper will review the most important aspects of hematopathological diagnostics in MDS, risk scoring, and their application to the inclusion and stratification of patients into the European Organization for Research and Treatment of Cancer (EORTC)/German MDS Study Group Phase III multicenter trial of low-dose decitabine in patients more than 60 years old with high-risk MDS. Emphasis is placed on itemizing the broad spectrum of cytologic and histologic stigmata defining the myelodysplastic categories that are to be considered in this study.

Antimetabolites, Antineoplastic↗
PubMed1999

Paraffin immunohistochemistry as an adjunct to hematopathology.

In the past two decades, there have been a number of dramatic advances in immunology that have subsequently affected immunohistochemistry. These have had a substantial impact on all phases of laboratory medicine, especially surgical pathology. Many hospital laboratories use immunohistochemical analysis on paraffin-embedded tissue sections as routinely as other special histochemical techniques. Hematopathology, in particular, has become increasingly reliant on the immunohistochemical antibody technique. This method aids in the identification of lineage-associated or lineage-restricted antigens in hematolymphoid neoplasms. The demonstration of kappa or lambda light-chain restriction is a good indicator of monoclonality in B-cell processes, aiding in the distinction between benign reactive hyperplasia and malignant lymphoma. Increasing numbers of antibodies that react to cluster differentiation cell surface antigens have become available for use in paraffin-embedded sections, obviating the need for frozen tissue in many cases. This facilitates the collection, processing, shipping, and storage of tissue specimens for immunohistochemical studies. This report will discuss the basic concepts in the immunohistochemical technique and show its applicability to hematopathology. It will also describe the most relevant antigens/antibodies and how they are used in antibody panels for the diagnosis of benign and malignant hematolymphoid lesions.

Antibodies, Neoplasm↗
PubMed2007

New transcription factors in diagnostic hematopathology.

The transcription factors (TFs) that controls the intricate machinery of multistep differentiation and activation programs of the lymphoid system, represent a complex array of proteins, whose identification and function has only in part been completed. TFs are usually expressed during specific differentiation or activation cellular programs, making them interesting tools in diagnostic immunohistochemistry. In fact, the specificity of some of these TFs for lineage or activation/differentiation passages or their abnormal expression in specific disease entity, represents a feature that has been exploited in diagnostic/prognostic immunohistochemistry. Bcl-6 was the prototype of this class of markers. Currently, the expanding knowledge of the TFs involved in the differentiation programs and in the activation processes of T-lymphocyte and B-lymphocyte in normal and neoplastic conditions and the availability of antibodies able to efficiently recognize these TFs in histologic material, represent a powerful tool in diagnostic hematopathology. In this review we will consider the basic biologic aspects and the applications in hematopathology of some of the lymphocyte-related TFs, including Pax5/BSAB, MUM1/IRF4, BOB1, Oct-2, T-bet, and FOXP3. This field is rapidly evolving, as witnessed by the ongoing growing number of novel TFs with possible diagnostic applications appearing in the literature.

Biomarkers↗
PubMed2004

Role of flow cytometry in pediatric hematopathology.

Multiparameter flow cytometry with optimally selected antibody combinations has expanded the use of this technique beyond traditional applications in hematopathology. By analyzing qualitative patterns of antigen expression on discrete populations or "clusters," one can detect immunophenotypic aberrancy in specific cell populations relative to normal and reactive populations. Evaluation of patterns of antigen expression can also be used to supplement conventional methodologies in the diagnosis and subclassification of certain types of hematologic neoplasia. Finally, the diagnosis of some congenital disorders affecting the hematolymphoid system can be facilitated by the detection of characteristic immunophenotypic changes. Examples of applications of multiparameter flow cytometry with particular relevance to pediatric hematopathology are discussed.

Burkitt Lymphoma↗
PubMed1990

In situ hybridization in hematopathology.

In situ hybridization (ISH) is one of the molecular techniques that has applications in diagnostic hematopathology. This procedure allows the detection of DNA or RNA in intact cells from various preparations, including cytology specimens and routinely fixed paraffin-embedded tissues. ISH is therefore analogous to detecting proteins (antigens) in intact cells with immunohistochemistry. The purpose of this article is to review the basic concepts and principles of ISH and to briefly discuss the important technical details of this procedure. Examples of potential applications of ISH in hematopathology are then discussed, including detection of Epstein-Barr virus, Y chromosome, and oncogene activation.

Autoradiography↗
PubMed1987

The hematopathology of cefonicid- and cefazedone-induced blood dyscrasias in the dog.

Cephalosporin treatment in man has been associated with blood dyscrasias that include a time- and dose-related anemia, neutropenia, and thrombocytopenia, the hematopathology of which remains poorly characterized. A similar hematologic syndrome can be produced in dogs following daily intravenous injections of 540-840 mg/kg cefazedone or 400-500 mg/kg cefonicid for 1-3 months. Using this animal model, histologic and cytologic changes in blood, bone marrow, spleen, and liver were studied over the course of the cephalosporin-induced cytopenias. Peripheral blood cytologic observations included an absence, generally, of erythroid regenerative changes, increased numbers of macroplatelets, spherocytosis, erythroblastemia, and toxic neutrophil morphology. Interim and postmortem cytologic and histologic observations of bone marrow included hypoplastic and toxic changes, primarily in cytopenic dogs receiving high doses of cefonicid, and regenerative changes in hematopoietic tissue of affected cefazedone-treated animals. The latter included variable erythroid hyperplasia, increased megakaryocytes, and decreased marrow fat and was accompanied by evidence of extra-medullary hematopoiesis and increased hemosiderin and hemophagocytosis in liver and splenic tissue. The incidence and severity of these changes were dose-dependent, corresponded with the cytopenias observed peripherally, and, like the cytopenias, were fully reversible. These observations suggest that the hematologic syndrome associated with cephalosporin treatment in the dog has multiple toxicologic mechanisms, which include peripheral cytotoxic effects and bone marrow damage with depressed or ineffective hematopoiesis.

Animals↗
PubMed2005

Standardization of bone marrow features--does it work in hematopathology for histological discrimination of different disease patterns?

Standardized bone marrow (BM) features determined by semiquantitative scoring are valuable tools for the recognition and easily reproducible interpretation of histological patterns in hematopathology. This procedure may help to characterize various disease entities, but especially to differentiate chronic myeloproliferative disorders (MPDs) with increased platelet counts from reactive thrombocytosis (RTh). A clear-cut separation of these conditions continues to present a major problem in hematology. Therefore MPDs are a most suitable model to test the diagnostic relevance of this procedure. By regarding the literature and based on archive material that involved BM biopsies of 319 patients, a semiquantitative grading of histological parameters was performed. Standardized features were applied for a stepwise discriminant analysis to establish different sets of variables exerting a diagnostic impact. A distinction into five histological patterns was achieved that showed a correctly predicted group membership of about 94 %. These were consistent with the clinicopathological diagnosis of polycythemia vera, essential thrombocythemia (ET), prefibrotic or early fibrotic chronic idiopathic myelofibrosis (CIMF) and finally RTh. Variables of discriminating potency according to their ranking included megakaryopoiesis (maturation defects, nuclear lobulation, naked and bulbous nuclei, small and giant size), reticulin fibers, erythro- and granulopoiesis (left shifting and quantity) and cellularity. These findings are in keeping with the assumption that characteristic patterns of BM histopathology can be assigned to different subtypes of MDPs mimicking ET. Discrimination between ET and especially early stage CIMF with thrombocythemia is warranted because of significant implications concerning therapeutic strategies, follow-up examinations and survival. Regarding these results, a schematic procedure is proposed to be used for daily routine diagnosis concerning the discrimination of MPDs.

Bone Marrow↗
PubMed1999

Hematopathology: integration of morphologic features and biologic markers for diagnosis.

The Revised European-American Classification of Lymphoid Neoplasms (REAL) represented a new paradigm for the classification of lymphomas. Although earlier classification schemes relied strictly on morphologic features or immunophenotype for the definition of diagnostic categories, the REAL classification emphasized that each disease was a distinct entity, defined by a constellation of clinical and laboratory features, i.e., morphologic and genetic features, immunophenotype, clinical presentation, and course. It also noted that the site(s) of presentation were often a signpost for important underlying biologic distinctions. The inclusion of clinical criteria for the definition of disease entities was one of the more novel aspects of the REAL classification. These principles of classification of lymphomas and leukemias also have an impact on diagnostic hematopathology. The REAL classification stresses that diagnoses are based on a multiparameter approach. Although morphologic features alone might suffice for some diagnoses, diagnostic accuracy and reproducibility are enhanced when immunophenotypic and genetic studies are also used. In addition, adequate clinical information is essential for some diagnoses, in particular the mature T-cell lymphomas and leukemias. The proposed World Health Organization classification of lymphoid and hematopoietic neoplasms incorporates these principles of classification and diagnosis.

Antigens, Neoplasm↗
PubMed1990

Molecular gene rearrangement analysis in hematopathology.

The advent of monoclonal antibody technology revolutionized the understanding of lymphoid and myeloid differentiation and greatly enhanced the ability to make a more accurate diagnosis. However, certain limitations became evident as monoclonal antibodies were increasingly applied to diagnostic hematologic problems. The T-cell antigens identified by monoclonal antibodies are not true clonal markers. B-cell immunoglobulin light chain restriction is a true clonal marker, but immunoglobulin is not expressed by many B-cell malignancies, diminishing its usefulness as a clonal marker. The development of molecular biologic techniques to augment diagnosis of hematologic neoplasms has revolutionized the understanding of neoplasia. Gene rearrangement analysis yields information concerning clonality and lineage. Gene rearrangement analysis can also identify chromosome breakpoints and translocations that correlate with specific diseases or prognoses. The purpose of this review is to familiarize the reader with the current applications of molecular biology to hematologic processes and to chart future directions of the application of these powerful techniques to diagnostic hematopathology.

DNA↗
PubMed1997

Posttransplant lymphoproliferative disorders: summary of Society for Hematopathology Workshop.

Twenty cases of posttransplant lymphoproliferative disorders arising in solid organ allograft recipients (18 patients) or bone marrow allograft recipients (2 patients: 1 autologous; 1 allogeneic) were selected for presentation at the Society for Hematopathology Workshop. In the course of the Workshop discussions, based both on the submitted cases and the combined experience of the participants, it was possible to agree on several distinctive categories of PTLD. These include (1) early lesions, (2) polymorphic posttransplant lymphoproliferative disorders (PTLDs), (3) monomorphic PTLDs (B- and T-cell lymphomas), (4) plasmacytoma-like lesions, and (5) T-cell-rich large B-cell lymphoma/Hodgkin's disease-like lesions. Monomorphic lesions should be classified according to a recognized classification of non-Hodgkin's lymphoma, although specified in the report as PTLD. Polymorphic lesions should be carefully evaluated for clonality; by immunophenotyping; and, if necessary, analysis of antigen-receptor and Epstein-Barr virus (EBV) genomes. Minimal pathological evaluation should include routine morphology, immunophenotyping on fresh tissue (flow cytometry or frozen section), and preservation of tissue for molecular genetic analysis. Analysis of the presence of EBV can be useful in establishing whether early or equivocal lesions represent PTLD (EBV+) or unrelated processes, but is not required in most cases. The pathologist can make an important contribution to the management of patients with PTLD by providing a complete diagnostic evaluation of the biopsy specimens (this is the least expensive part of the care of a transplant patients, not a place to try to cut costs) and making sure the attending physicians understand the special issues in management of PTLD.

Bone Marrow Transplantation↗
PubMed1982

Systemic mast cell disease: a clinical and hematopathologic study of 26 cases.

Systemic mast cells disease (SMCD) is an uncommon disorder that constitutes approximately 10% of all mastocytoses. Diagnosis requires a substantial degree of clinical suspicion, which may not be present if characteristic skin lesions of urticaria pigmentosa are not observed. Lack of well-defined histopathologic features for the disease have delayed or prevented the diagnosis of SMCD. An initial diagnosis of "myeloproliferative disorder," chronic granulocytic leukemia, or myelofibrosis is frequently made. Study of the clinical and pathologic features of 26 cases of SMCD indicated that affected patients are generally middle aged and may have had urticaria pigmentosa for many years. Gastrointestinal symptoms are common, and splenomegaly and hepatomegaly along with radiographic evidence of generalized bone disease are usually noted. Hematologic factors are highly variable. Characteristic histopathologic features of SMCD are described for bone marrow, lymph nodes, liver, and spleen. The authors consider tissue fixation and staining methods to help identify mast cell lesions.

Adult↗
PubMed2002

[Molecular diagnosis in hematopathology].

The diagnosis of hematopoietic neoplasias has nowadays become more and more refined by molecular cytogenetic and molecular biological techniques. The biological basis of most of these techniques is that (malignant) tumors represent clonal neoplasias that are derived from a single or few progenitor cells, thus imparting a clonal relationship to all daughter cells. Apart from clonality analysis as a marker for neoplastic growth, the presence or absence of certain primary chromosome abnormalities, and the number and the kind of secondary genetic anomalies become increasingly important in the delineation of the biological grade of aggressiveness.

Biomarkers, Tumor↗
PubMed2000

[Myelodysplastic syndromes (MDS). Aspects of hematopathologic diagnosis].

Myelodysplastic syndromes (MDS) are a heterogenous group of clonal stem cell disorders which generally occur in older adults but may also affect children. Primary MDS should be distinguished from secondary MDS associated with antineoplastic or immunosuppressive therapy (t-MDS), exposure to toxic compounds, or genetic disorders. The establishment of a neoplastic clone is reflected by dysplastic features and impaired function which may affect all three hematopoietic cell lineages. The ineffective hematopoiesis which causes bone marrow failure is accompanied by peripheral blood cytopenia and is considered to result from increased apoptosis, at least in the less advanced MDS stages. The elucidation of the molecular pathogenesis of MDS has provided evidence that chromosomal abnormalities are present in about 50% of patients with primary MDS. They include numerical aberrations such as monosomy 5 or 7, trisomy 8, loss of the Y-chromosome and structural abnormalities such as deletion of the long arm of chromosome 5 (5q-syndrome), 7, or 8. Based on the percentage of blasts (<5%, 5-20%, 20-30%) and the presence of >15% ringed sideroblasts for marrows with <5% blasts, the French-American-British (FAB) classifies MDS into 4 morphologic categories: refractory anemia (RA), refractory anemia with excess of blasts (RAEB), refractory anemia with excess of blasts in transformation (RAEB-t), and refractory anemia with ringed sideroblasts. The fifth morphologic type is chronic myelomonocytic leukemia characterized by peripheral blood monocytosis (>1x10(9)/l). However, a modification of this classification will be proposed by the World Health Organization, with the intention of lowering the threshold for the diagnosis of AML from 30% to 20% blast cells. In patients presenting with cytopenias suggesting impaired hematopoiesis, the initial diagnosis depends mainly on the cytological evaluation of bone marrow and blood smears and the histological findings of trephine bone marrow biopsy. In a retrospective analysis we evaluated the occurrence of the distinct FAB-categories as percentage of the total number of MDS cases diagnosed at the Institute of Pathology of the University of Freiburg. A total of 63% fullfilled the criteria of RA/RARS, 17% of RAEB, 14% of RAEB-t, and 6% of CMML. A fibrotic variant of MDS was observed in 7.67% of all cases, ranging from 2.34% in RA up to 15. 42-15.84% in the categories which did not show significant differences with regard to myelofibrosis. The histologic evaluation of a trephine bone marrow biopsy is of critical importance for the evaluation of fibrotic or hypocellular MDS since these patterns are not reflected by the cytological examination. The combined cytological and histological diagnosis of bone marrow and peripheral blood is a reliable tool for the initial diagnosis of MDS. In addition, cytogenetic and molecular analysis should be performed. Presently, the risk of leukemic transformation is evaluated using the International Prognostic Scoring System for MDS, which is the sum of the scores of bone marrow blasts, karyotypes and cytopenia. In the context of clinical trials therapeutic modalities should be considerd according to the age and the general performance state and the prognostic scores of individual patients.

Adult↗