Standardizing immunophenotyping for the human immunology project. Foon2 1division of hematopathology, department of pathology, and 2division of hematologyoncology, department of medicine, university of pittsburgh school of medicine, pa flow cytometric immunophenotyping remains an indispensable tool for the diag. Leukemia and lymphoma immunophenotyping represent an extremely important complement to morphology in the diagnosis and monitoring of hematopoietic malignancies. The most common use of flow cytometry is in the identification of markers on cells, particularly the immune system or immunophenotyping. Immunophenotyping refers to the technique of identifying molecules that are associated with lymphoma cells and that help to characterize them. Flow cytometric immunophenotyping for hematologic neoplasms.
The acs laboratory assessment of leukaemia lymphoma immunophenotyping is an acs document to be read in conjunction with acs documents guidelines for clinical flow cytometry laboratory practice and guidelines for leukaemia lymphoma immunophenotyping. Application of flow cytometry in the diagnosis a leukemia and lymphoma german. Cytometry, leukemia immunophenotyping flow cytometry, lymphoma immunophenotyping leukemia immunophenotyping lymphocytoflow lymphoma immunophenotyping by flow cytometry plasma cell. Lymphoma is the general term for many different types of blood cancer. Greater manchester cancer pathway lymphoma guidelines. Immunophenotyping by flow cytometry lab tests online.
Fcs files, from which central analyses manual and automated were performed, and. Immunophenotyping and flow cytometry test guidelines test ordering for immunophenotyping and flow cytometry is based on the patients diagnosis andor disease state. Nonhodgkin lymphoma and hodgkin lymphoma are the two main types. Rapid, effective and userfriendly immunophenotyping of. Greater manchester cancer pathway lymphoma guidelines 2019. It is the desire to detect an increased number of cell subsets of the immune system that has driven the increase in multicolor flow cytometry. Flow cytometric immunophenotyping of adult tcell leukemia lymphoma using cd3 gating taiji yokote, md,1 toshikazu akioka, md,1 satoko oka, md,1 satoshi hara, md,1 kichinosuke kobayashi, md,1 hideto nakajima, md,1 takeshi yamano, md,1 toshiyuki ikemoto, phd,3 akira shimizu, md,3 motomu tsuji, md,2 and toshiaki hanafusa, md1. The european leukemianet eln, workpackage 10 wp10 was designed to deal with diagnosis matters using morphology and immunophenotyping. Anaplastic large cell lymphoma alcl is usually diagnosed by histologic and immunohistochemical analysis.
In addition, flow cytometric immunophenotyping fci has become a widely used laboratory procedure for diagnosis and subtyping of lymphoma. Sample cases click here for instructions on how to download the free fcs express reader to view and manipulate the sample cases. Immunophenotypic criteria for the diagnosis of nonhodgkin. Immunophenotyping cd marker labeling pbmcs with orflos moxi go next generation flow cytometer histogram for that region. To evaluate the role of flow cytometric immunophenotyping fci in diagnosis and characterization of lymphoma tissue specimens from egyptian patients. Peripheral blood cd2, cd19, cd20, cd3, cd4cd3, cd8cd3, helper suppressor ratio. The test results indicate whether the persons lymphocytes are derived from either a single cancer cell leukemia or from other noncancerous conditions. Evaluating leukemia and lymphoma in blood, marrow and tissue, where it can be essential for accurate diagnosis. Immunophenotyping of thymic lymphoma in a nelore cow. Using a method of analysis relying solely on immunoarchitectural features of a given case.
In the context of lymphoma diagnosis, immunohistochemistry ihc has the. Diagnosis recommended tests epic code acute leukemia all, aml, amml, cmlbc. It provides a lot of useful information in this setting that transfers directly from laboratory to clinical management of patients. Lymphocyte immunophenotyping is an important test used to diagnose cll and other types of leukemia and lymphoma, by comparing the cancer cells to normal immune cells. Traditionally, 23 of dogs with lymphoma are classified as b cell and are t cell. Immunophenotyping of acute leukemias from biology to. The diagnosis of hodgkin lymphoma hl is defined in terms of its microscopic appearance and the expression of cell surface markers. Immunophenotyping of classical hodgkins lymphoma by flow cytometry. Flow cytometric immunophenotyping of nonhodgkins lymphomas and related disorder.
Tissue except lymph node, bronchial alveolar lavage, sterile. To address this issue, we performed immunophenotyping of pdl1positive cells in hodgkin lymphoma and in angioimmunoblastic t cell lymphoma aitl employing multiple immunofluorescent immunolabeling. In veterinary medicine, currently canine lymphoma is broken down into two main immunophenotypes, b and t cell. Third, ageing has been associated with an increase in the number of cd28 latestage effector t cells16 and an increase in the number of oligoclonal cytomegalovirusspecific t cells 17. Angioimmunoblastic tcell lymphoma aitl is a distinct form of peripheral tcell lymphoma ptcl frequently involving lymph nodes, spleen and bone marrow, and is associated with systemic symptoms. Our report also documents the unusual expression of t. Review article flow cytometric immunophenotyping for hematologic neoplasms fiona e. The cell subsets prespecified for evaluation in the study are shown in table 2.
Acs guideline for haematology oncology immunophenotpying v scope the acs guideline for leukaemia lymphoma immunophenotyping is an acs document to be read in conjunction with the acs document guidelines for clinical flow cytometry. Leukemialymphoma immunophenotyping profile labcorp. Discrepancies in the immunophenotype of lymphoma cells in. Immunophenotypic criteria for the diagnosis of nonhodgkins.
Test information immunophenotyping and flow cytometry. Immunophenotyping of diffuse large bcell lymphoma dlbcl. Study have successfully developed standardized immunophenotyping panels and procedures for leukemia and lymphoma diagnostics and whole blood immunophenotyping, respectively. A clinical study ashish gupta1, abhijit pal1, silas supragya nelson2 1assistant professor, department of medicine, nscb medical college, jabalpur, madhya pradesh, india, 2associate professor, department of medicine, nscb medical college, jabalpur, madhya pradesh, india meaning white blood. Methods cases from 1988 to 2009 with an fna diagnosis of thyroid lymphoma were selected from hospital records. Immunophenotyping for leukemia lymphoma, tissue, bronchial alveolar lavage, or sterile body fluid gipp. Immunohistochemistrylymphomanonhodgkin lymphomahodgkin lymphomaheatinduced epitope retrieval monoclonal antibody. The legends summarize the diagnosis, the threecolor tube format fitc, pe, peridinin chlorophyll percp, or cy51, and the purpose of the illustrations. Validation and quality control of immunophenotyping in. Fci using 2 and 3 color staining approaches, was performed on 50 fresh lymph nodes specimen from cairo nci patients with suspected lymphoma presenting with either localized or generalized lymphadenopathy. We found that pdl1positive cells and pd1positive cells both in hodgkin lymphoma and in aitl belong to two completely different cell lineages. Guidelines on diagnosis and treatment of malignant.
Immunophenotyping by flow cytometry is a laboratory method that may be used to help diagnose and classify a leukemia or lymphoma. Immunophenotyping cd marker labeling pbmcs with orflos. To evaluate the role of flow cytometric immunophenotyping fci in diagnosis and. Flow cytometric analysis of leukemia and lymphoma the basics. This is our first attempt to bring some uniformity in reporting of hematolymphoid neoplasms in india. It is essential that such an approach underpins the clinical management of these diseases, many of which are amenable to cure. Diffuse large bcell lymphoma dlbcl forms a heterogeneous collection of aggressive nonhodgkins lymphoma in which three principle classes of neoplasia have been defined according to gene expression and immunophenotyping studies.
Standardizing flow cytometry immunophenotyping analysis from. Guidelines on diagnosis and treatment of malignant lymphomas diagnosis. Immunophenotyping leukemias and lymphomas click here to start. Tissue flow cytometry immunophenotyping in the diagnosis and. The role of fineneedle aspiration in the diagnosis of. If both tissue flow cytometry and histology are required, submit one portion of fresh specimen in transport medium or saline for flow. Immunophenotyping of nonhodgkins lymphomas in sudan. Hodgkin lymphoma diagnosis leukemia and lymphoma society. Immunophenotyping is a process of classifying cells of the immune system based on structural and functional differences.
These studies used a similar approach by assessing correlation and agreement with linear. This can be done on tissue section fresh or fixed tissue, cell suspension, etc. An electronic medliner and embase search retrieved. The role of immunophenotyping in differential diagnosis of. This process is used to diagnose specific types of leukaemia and lymphoma by comparing the cancer cells to normal cells of the immune system. Flow cytometry fc immunophenotyping has become a standard practice in the. Illustrative examples of lymphoma immunotyping by threecolor flow cytometry. Evaluating lymphocytoses of undetermined etiology identifying b and tcell lymphoproliferative disorders involving blood and bone marrow distinguishing acute lymphoblastic leukemia all from acute myeloid leukemia aml immunologic subtyping of all distinguishing reactive lymphocytes and lymphoid hyperplasia from malignant lymphoma. Flow cytometry is one of the most powerful tools for singlecell analysis of the immune system at a cellular level. Acs guideline for haematology oncology immunophenotpying v scope the acs guideline for leukaemia lymphoma immunophenotyping is an acs document to be read in conjunction with the acs document guidelines for clinical flow cytometry laboratory practice. Flow cytometric immunophenotyping of anaplastic large cell. A process used to identify cells, based on the types of antigens or markers on the surface of the cell. The molecules are analyzable because in most cases they are expressed on the outer cell surface membrane. With that view, the fine sizediscrimination of the impedancebased detection approach is clearly highlighted in the resolution.
Immunophenotyping of classical hodgkins lymphoma by flow. Immunophenotyping uses antibodies to identify white blood cells based on the types of antigens or markers on their surface or their interior. Immunophenotypic criteria for the diagnosis of nonhodgkins lymphoma. Standardizing flow cytometry immunophenotyping analysis. About marrow, blood and blood cells the information in this section about normal blood and marrow may help you understand the lymphoma information in. Other tests will be done to make sure the diagnosis of all is correct. Aufzahlung, other related material on this website in german, pdffiles. Generating an epub file may take a long time, please be patient. Immunophenotyping of angioimmunoblastic tcell lymphomas.
Also, in many instances, although a specific subclassification may have a predominant immunophenotypic pattern, there is often variability with markedly different immunophenotypic subsets even within a given. Diagnostic hematopathology has become an increasingly complex subspecialty, particularly with neoplastic disorders of blood and bone marrow. While morphologic assessment of blood smears, bone marrow smears, and tissue sections remains the cornerstone of lymphoma and leukemia diagnosis and classification, immunophenotyping is a very valuable and important complementary tool. Diagnosis recommended tests epic code acute leukemia all, aml, amml, cmlbc leukemialymphoma immunophenotyping profile.
However, fineneedle aspiration is becoming a popular alternative to lymph node biopsy, and flowcytometric immunophenotyping is often used to analyze fineneedle aspiration specimens. Pdf immunophenotyping of thymic lymphoma in a nelore cow. Department of internal medicine, university hospital, utrecht, the netherlands. In order to classify the cell types of interest, detecting multiple antigens on the cells is often necessary. 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 nonhodgkins lymphoma.
Foon2 1division of hematopathology, department of pathology, and 2division of hematologyoncology, department of medicine, university of pittsburgh school of medicine, pa flow cytometric immunophenotyping re. Diffuse large bcell lymphoma dlbcl the single largest category of lymphoma, is a heterogeneous group of malignant lymphomas with several morphologic features. In a retrospective analysis the authors studied the relation between the immunologic phenotype of bcell nonhodgkins lymphoma nhl and diseasefree survival. Flow cytometric immunophenotyping for hematologic neoplasms fiona e. Following the course of a disease and may identify relapse or persistent disease in. It is an objective and quantitative diagnostic tool that allows quick multiparametric analysis of a very large number of cells 20. Oct 26, 2012 in our experience, while the neoplastic population the hodgkins reedsternberg cell of classical hodgkins lymphoma chl cannot be detected by flow cytometry assays designed to detect b and t cell non hodgkins lymphoma, six and ninecolor reagent combinations are available to detect these cells with high diagnostic sensitivity and specificity. It is also suitable for investigating paucicellular samples such as cerebrospinal fluid or fine needle aspirates. A hematopathologist is a specialist who studies blood and bone marrow cells and other tissues to help diagnose diseases of the blood, bone marrow and lymph system. Standardizing flow cytometry immunophenotyping analysis from the human immunophenotyping consortium.
The hematopathologist looks for the presence of certain antigens or markers on the surface of the cells in order to identify hodgkin and rs cells and confirm a diagnosis of hl. Immunophenotyping lymphocyte subsets flow contract site. Flow cytometric immunophenotyping of adult tcell leukemia. Sumeet gujral principal investigator, icmr task force. Immunophenotyping this test is used to identify cells based on the types of proteins on the cell surface, to see which of the main subtypes of all you have, bcell all or tcell all.
Guidelines on diagnosis and treatment of malignant lymphomas. Lineage definition is the first goal leading to proper initial therapy. Immunophenotyping is a technique to analyze the immune cell subsets by detecting proteins expressed on the surface or within the cell intracellular using antigen specific antibodies. Pdf more than two decades have past since the recognition of. Various approaches in the histological classification of hl have been developed to predict prognosis on a histomorphological basis. Immunophenotyping is a technique used to study the protein expressed by cells.
The files of the flow cytometry laboratory, department of hematopathology, m. A technique called immunophenotyping is used to distinguish hl from other types of lymphoma or other cancerous or noncancerous conditions. Immunophenotyping is an essential part of the modern diagnostic workup of acute leukemias and thus for an appropriate treatment of these complex and heterogeneous diseases. Standard operating procedures for immunophenotyping of.
Keywords lymphocyte, lymphoma, multiparameter flow cytometry, screening 1 introduction in 2014, approximately 5000 specimens from peripheral blood. Sumeet gujral principal investigator, icmr task force project. Following the course of a disease and may identify relapse or persistent disease in individual cases. These documents outline guidelines for good medical pathology.
Pdf thymic lymphoma is a malignant lymphoid neoplasm that affects several species, including cattle. You may want to get a second medical opinion by an experienced hematopathologist before you begin treatment. Fcs files, from which central analyses manual and automated were performed, and instrument setup parameters. Here we demonstrate that an automated data analysis strategy can be integrated into a workflow utilizing a standardized staining panel. Pdf flow cytometric immunophenotyping of nonhodgkins. This technique is commonly used in basic science research and laboratory diagnostic purpose. Molecular oncology cytogenetics and fish aided by t or bcellspeci. This group aimed at establishing a consensus on the. Our finding of costeffective, accurate and rapid immunophenotyping of canine lymphoma using the personal flow cytometer is similar to the findings of other studies on immunophenotyping lymphocytes in hiv infection in humans 24, 31, 33, 34, 37, 38.
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