CD3, CD4, CD5, CD7, CD8, CD10, CD19, CD20, CD16 + CD56, CD23; kappa; lambda; additional markers added as needed (CD2, CD25, CD38, CD71, TRC alpha/beta, TCR gamma/delta)
Whole blood or bone marrow aspirate or body fluids
Two 7-mL EDTA whole blood tubes or one 7-mL EDTA whole blood tube and 2 cc EDTA or heparinized bone marrow aspirate. One EDTA whole blood tube is for an optional CBC. One 7-mL EDTA body fluid tube. Large volumes of body fluids should be concentrated and transferred to an EDTA tube.
Lavender-stopper (EDTA whole blood or body fluid) tube, lavender-stopper (bone marrow) tube or green-stopper (heparinized bone marrow) tube
Submit specimen at room temperature using Leukemia/Lymphoma Specimen Transport Kit (supplied by the Hospital). Draw specimen so it will arrive in the laboratory Monday through Saturday and within 24 hours of collection. Please state the date and time of collection on the test request form.
Maintain specimen at room temperature.
Causes for Rejection:
Specimen frozen; specimen in formalin or other fixative; clotted specimen; hemolysis; blood more than 72 hours old; bone marrow aspirates older than 5 days; bags or bottles of body fluid or bronchial washings
Identify and characterize the following:
1) reactive lymphocytosis vs chronic lymphocytic leukemia (CLL)
2) prolymphocytic leukemia vs lymphoblastic leukemia.
(large granular lymphocyte proliferations, T-gamma lymphoproliferative disease, natural killer
cell proliferations, T-cell CLL, T-cell gamma/delta proliferations)
3) Sézary syndrome
4) non-Hodgkin lymphoma (in blood, bone marrow, or body fluid)
5) adult T-cell leukemia/lymphoma
Genotyping, to detect T-cell receptor gene rearrangements, is recommended when the immunophenotyping profile suggests a clonal T-cell process. See T-Cell Receptor Gene Rearrangements for Leukemia.
Immunophenotyping by flow cytometry
Neoplastic B-cell proliferations (chronic leukemias and lymphomas) are clonal expansions of cells that express either kappa or lambda immunoglobulin light chains. In normal or reactive processes, a bimodal distribution of kappa- and lambda-positive B cells is present in a ratio of approximately 1.5:1. Immunophenotyping using multiparameter analysis (simultaneous staining with a pan B-cell marker and anti-immunoglobulin light chain antibodies) is a rapid and specific method for detecting and confirming the presence of neoplastic B-cell disorders.
Chronic lymphocytic leukemia (CLL) is a clonal lymphoproliferative disorder, usually of B-cell origin (95%) that has been traditionally diagnosed using clinical and morphologic criteria. Incorporation of immunophenotypic features into the diagnostic criteria is helpful in separating common B-cell CLL from other lymphoproliferative disorders. Detection of karyotypic abnormalities is useful in assessing prognosis. Lymphocytes in B-CLL coexpress CD19, CD20, and CD23 pan B-cell antigens, CD5, pan T-cell antigen, and a single immunoglobulin light chain, kappa or lambda. CD10 (CALLA) expression is usually absent. Mantle cell lymphoma is distinguished from CLL by absent or very dim expression of CD23.
T-cell CLL, unlike B-CLL, is associated with rapid onset, an aggressive clinical course poorly responsive to therapy and decreased survival. Immunophenotyping, in the majority of cases, demonstrates expression of CD3 (a pan T-cell antigen), and CD4 (the helper cell antigen). CD8 (the suppressor/cytotoxic cell antigen) is usually not expressed. Genotyping demonstrates a clonal rearrangement of the T-cell receptor gene.
Large granular lymphocyte (LGL) proliferations can be divided into T-cell and natural killer (NK) cell subsets by immunophenotyping. The more common T-cell type expresses CD3, a pan T-cell antigen and CD8, the suppressor/cytotoxic cell antigen. Genotyping demonstrates a rearrangement of the T-cell receptor gene. The NK cell type is relatively rare and expresses CD2 and CD16 and/or CD56. CD3 expression is absent. Genotyping demonstrates a germline configuration of the T-cell receptor gene.
In Sézary syndrome, the neoplastic lymphocytes are T cells with a helper cell phenotype. Expression of CD7, a pan T-cell antigen, is absent and is useful in distinguishing the neoplastic cells from normal T-helper cells. Genotyping demonstrates a clonal rearrangement of the T-cell receptor gene.
In adult T-cell leukemia/lymphoma, the neoplastic lymphocytes are T cells with a helper cell phenotype. Expression of CD3, CD4, and CD25 is present. Expression of CD7 is absent. Genotyping demonstrates a clonal rearrangement of the T-cell receptor gene.