Leukemia/Lymphoma Immunophenotyping Profile

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Test Details

Synonyms

  • Chronic Leukemia/Lymphoma
  • Flow Cytometry
  • Flow Immunophenotyping
  • Leukemia
  • Leukemia Profile
  • Lymphoma

Use

Identify and characterize the following:

• Reactive lymphocytosis vs chronic lymphocytic leukemia (CLL)

• CLL vs mantle cell lymphoma

• Prolymphocytic leukemia vs lymphoblastic leukemia large granular lymphocyte proliferations, T-γ lymphoproliferative disease, natural killer cell proliferations, T-cell CLL, T-cell γ/δ proliferations

• Sézary syndrome

• Non-Hodgkin lymphoma

• Adult T-cell leukemia/lymphoma

Neoplastic B-cell proliferations (chronic leukemias and lymphomas) are clonal expansions of cells that express either κ or λ immunoglobulin light chains.

Methodology

Immunophenotyping by flow cytometry

Additional Information

In normal or reactive processes, a bimodal distribution of κ- and λ-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, κ or λ. CD10 (CALLA) expression is usually absent. Mantle cell lymphoma is distinguished from CLL by absent or very dim expression of CD23.

Lymphomas are biologically complex neoplasms of the immune system. Numerous classification schemes have been developed based on morphologic features. This limited approach is often unreliable. Immunophenotyping, by flow cytometry and/or immunohistochemistry, has emerged as a valuable adjunct to conventional morphologic diagnosis and classification. Flow cytometry offers the advantage of rapid multiparameter analysis. Combining light scatter characteristics with patterns of antigen expression and DNA content provides biological information that is useful in making a diagnosis and assessing prognosis. Various gating strategies can be employed to enhance the detection of minor populations, thus providing a level of sensitivity comparable to molecular methods (gene rearrangement studies).

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.

Detection of a B-cell population coexpressing CD22, CD11c, and CD25 is useful in establishing a diagnosis of hairy cell leukemia when used in conjunction with morphology and cytochemistry. Immunophenotyping by flow cytometry is a sensitive method for detecting residual or recurrent disease in the peripheral blood of patients with an established diagnosis.

Detection of a population of cells expressing CD38 and CD138 in the peripheral blood is useful in establishing a diagnosis of plasma cell leukemia when used in conjunction with morphology.

Lineage assignment in acute leukemia is necessary for selecting appropriate therapy and is useful in assessing prognosis. Multiparameter analysis using four-color immunophenotyping techniques is a rapid and specific method of assigning lineage in acute leukemia.

This profile is also useful in distinguishing lymphoid from myeloid blast crisis in CML and immunophenotyping lymphoblastic lymphoma in blood or bone marrow. Immunophenotyping and cytogenetic analysis are increasingly being used to supplement the traditional methods (morphology and cytochemistry) of classifying acute leukemias and to provide prognostic information. Acute lymphoblastic leukemia (ALL) can be classified into undifferentiated null T- and B-cell lineages. In all of B-cell lineage, expression of CD10 (CALLA) is a favorable prognostic factor. Acute myelogenous leukemias (AML) are a heterogeneous group. In cases where morphology and cytochemical staining is equivocal, immunophenotyping can be useful. Immunophenotyping is particularly useful in classifying megakaryoblastic leukemia (FABM7). A combination of characteristic light scattering properties and myeloid phenotype can suggest a diagnosis of acute promyelocytic leukemia (FABM3). Confirmation of the retinoic acid receptor gene rearrangement by cytogenetic or molecular methods is recommended. See tests listed under Related Information and related FISH tests (eg, 510669).

Specimen Requirements

Specimen

Whole blood, bone marrow aspirate, body fluids, fresh lymph node, spleen, extranodal solid tissue, biopsy, or needle aspirate

Volume

3 mL whole blood or 2 mL bone marrow aspirate, 2 mL body fluid tube. Large volumes of body fluids should be concentrated to <5 mL; 0.5 to 1.0 cm3 fresh tissue.

Minimum Volume

1 mL whole blood or bone marrow (Note: This volume does not allow for repeat testing.)

Container

Green-top (sodium heparin) tube (preferred), lavender-top (EDTA) tube, or yellow-top (ACD) tube for whole blood or bone marrow (acceptable, not preferred); lavender-top (EDTA body fluids) tube; fresh tissue in lymph node transport bottle containing RPMI

Collection

Note: In an attempt to maintain specimen at room temperature,

• In hot weather, enclose a refrigerated (not frozen) gel pack in shipper kit.

• In cold weather, run hot water over gel pack for three to five minutes and enclose in shipper kit.

• Ship specimens in a timely manner based on the specific requirements of the test.

Submit blood or bone marrow at room temperature. Collect the specimen so it will arrive in the laboratory Monday through Saturday and within 24 hours of collection. Please state on the test request form the date and time of collection and the name and phone number of the pathologist responsible for the histologic or cytologic diagnosis.

For fresh tissue, aseptically cut tissue in pieces and place in lymph node transport bottle. If aspirate is submitted, rinse needle in transport medium. Submit at room temperature using Lymph Node Transport Kit (supplied by LabCorp). If transport kit is not available, place specimen in sterile container with saline. Submit specimen so it will arrive in the laboratory Monday through Saturday and within 24 hours of surgical removal. To avoid transportation delays, submit specimen on the day of collection.

Storage Instructions

Maintain specimen at room temperature.

Causes for Rejection

Hemolysis; specimen clotted; specimen frozen; specimen in formalin or other fixative; blood more than 72 hours old; bone marrow aspirates more than five days old; bags or bottles of body fluid or bronchial washing; tissue in formalin or other fixative; contaminated transport medium

Clinical Information

Special Instructions

If both tissue flow cytometry and histology are required, submit one portion of fresh specimen in transport medium or saline for flow cytometry and one portion in 10% formalin for histologic analysis.

Please direct any questions regarding this test to oncology customer service at 800-345-4363. Pathologist consultation is available Monday through Friday. Indicate differential diagnosis on test request form. Submit recent CBC results for consideration in report. Billing will be performed back end.

References

Bitter MA. Hairy cell leukemia. In: Knowles DM, ed. Neoplastic Hematopathology. Baltimore, Md: Williams and Wilkins;1992.
Braylan RC. Lymphomas. In: Bauer KD, Duque RE, Shankey TV, eds. Clinical Flow Cytometry: Principles and Applications. Baltimore, Md: Williams and Wilkins;1993.
Foucar K. B-Cell chronic lymphocytic and prolymphocytic leukemia. In: Knowles DM, ed.Neoplastic Hematopathology. Baltimore, Md: Williams and Wilkins;1992.
Knowles DM II. The human T-cell leukemias: Clinical, cytomorphologic, immunophenotypic, and genotypic characteristics. Hum Pathol. 1986; 17(1):14-33. 3002948
McDaniel HL, MacPherson BR, Tindle BH, et al. Lymphoproliferative disorder of granular lymphocytes: A heterogeneous disease. Arch Pathol Lab Med. 1992; 116(3):242-248. 1371379
Miller ML, Fishleder AJ, Tubbs RR. The expression of CD22 (leu 14) and CD11c (leu M5) in chronic lymphoproliferative disorders using two-color flow cytometric analysis. Am J Clin Pathol. 1991; 96(1):100-108. 2069128

LOINC® Map

Order Code Order Code Name Order Loinc Result Code Result Code Name UofM Result LOINC
480260 Comp panel: Leukemia/Lymphoma 490024 Flow Interpretation 50595-8
480260 Comp panel: Leukemia/Lymphoma 490025 Flow Comment 48767-8
480260 Comp panel: Leukemia/Lymphoma 490026 Dianon Case Number N/A
480260 Comp panel: Leukemia/Lymphoma 490045 Clinical Information 55752-0
480260 Comp panel: Leukemia/Lymphoma 490027 Specimen Type 31208-2
480260 Comp panel: Leukemia/Lymphoma 490028 Assessment of Leukocytes N/A
480260 Comp panel: Leukemia/Lymphoma 490031 Cellularity Assessment 67127-1
480260 Comp panel: Leukemia/Lymphoma 490032 Viability 33194-2
480260 Comp panel: Leukemia/Lymphoma 490033 Immunophenotypic Profile 67128-9
480260 Comp panel: Leukemia/Lymphoma 490034 PNH Flow Assay N/A
480260 Comp panel: Leukemia/Lymphoma 490035 Analysis and Gating Strategy N/A
480260 Comp panel: Leukemia/Lymphoma 490036 Phenotype Chart 55230-7
480260 Comp panel: Leukemia/Lymphoma 490039 Resulting Path Name 19139-5
480260 Comp panel: Leukemia/Lymphoma 490040 Clinician provided ICD9: 52797-8
480260 Comp panel: Leukemia/Lymphoma 490041 Resulting ICD9: 52797-8
480260 Comp panel: Leukemia/Lymphoma 490042 Indications for Study 42349-1
480260 Comp panel: Leukemia/Lymphoma 490046 Comment: 77202-0
480260 Comp panel: Leukemia/Lymphoma 480059 Flow LLS Tracking N/A

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