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Immunophenotyping

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Also known as: Immunophenotyping by Flow Cytometry or Immunohistochemistry
Formal name: Immunophenotyping

At a Glance

Why Get Tested?

To help diagnose and classify a leukemia or lymphoma; to help guide treatment; to detect and evaluate leukocyte cancer cells that remain after treatment or disease relapse

When to Get Tested?

When you have signs and symptoms that a health practitioner thinks may be due to leukemia or lymphoma; when you have been diagnosed with leukemia or lymphoma but the specific subtype is unknown; sometimes to evaluate the effectiveness of treatment or to evaluate for recurrent disease

Sample Required?

A blood sample drawn from a vein in your arm; sometimes a bone marrow, tissue biopsy, or fluid sample collected by a health practitioner

Test Preparation Needed?

None

The Test Sample

What is being tested?

Immunophenotyping detects the presence or absence of white blood cell (WBC) antigens. These antigens are protein structures found on the surface or interior of WBCs. Typical groupings of antigens are present on normal WBCs and are unique to specific cell types. Atypical but characteristic groupings are seen with leukemias and lymphomas. This allows immunophenotyping to be useful in helping to diagnose and classify these blood cell cancers.

Leukemias and lymphomas are caused by an abnormal cell that begins to clone itself uncontrollably. The abnormal lymphocytes or myeloid (granular) monoclonal cells proliferate, yet do not fight infections or perform other functions like normal WBCs. Because they do not die at a normal rate, they accumulate in the bone marrow, in a lymph node, or in other tissues, where they grow in numbers. As the number of cells increases in the bone marrow, they may crowd out and inhibit the production of normal red and white blood cells. Eventually, the abnormal cells will also be released into the bloodstream.

CBC (complete blood count) and differential tests performed on a sample of blood from someone with leukemia or a lymphoma may reveal an increased number of white blood cells with a predominance of one type. These tests may suggest lymphoma or leukemia, but more information is generally needed to confirm a diagnosis. CBC and differential testing cannot confirm the presence of monoclonal WBCs or detect the subtle differences that may exist between different types of blood cell cancers.

With immunophenotyping, a blood, bone marrow, or other tissue sample can be tested to gather this information – information that is then used to identify a specific type of leukemia or lymphoma and, where possible, used to predict its likely aggressiveness and/or responsiveness to certain treatment. The identification of different types of leukemias and lymphomas is based upon the presence or absence of antigens and a typical pattern that has been established with each leukemia/lymphoma.

Immunophenotyping detects the presence or absence of antigens found on the surface or interior of blood cells. Atypical or abnormal cells can demonstrate characteristic antigen groupings that are consistent with specific types of leukemia and lymphoma. The identifications made are based upon a "library" of antigen associations and patterns that have been established over time.

Most of the antigens that immunophenotyping detects are identified by a CD (clusters of differentiation or cluster designation) number (see the table in the "What does the test result mean?" section). CD numbers represent a naming convention that is based upon international consensus. While hundreds of antigens have been identified and have received a unique CD number, only a small number of these are routinely used.

How is the sample collected for testing?

A blood sample is obtained by inserting a needle into a vein in the arm. A bone marrow aspiration and/or biopsy procedure is performed by a trained health practitioner. Fluid samples are obtained through collection of the fluid in a container or by inserting a needle into the body cavity and aspirating a portion of the fluid with a syringe. Sometimes, a tissue sample, such as from a lymph node, is obtained using a biopsy procedure.

NOTE: If undergoing medical tests makes you or someone you care for anxious, embarrassed, or even difficult to manage, you might consider reading one or more of the following articles: Coping with Test Pain, Discomfort, and Anxiety, Tips on Blood Testing, Tips to Help Children through Their Medical Tests, and Tips to Help the Elderly through Their Medical Tests.

Another article, Follow That Sample, provides a glimpse at the collection and processing of a blood sample and throat culture.

Is any test preparation needed to ensure the quality of the sample?

No test preparation is needed.

The Test

Common Questions

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Article Sources

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NOTE: This article is based on research that utilizes the sources cited here as well as the collective experience of the Lab Tests Online Editorial Review Board. This article is periodically reviewed by the Editorial Board and may be updated as a result of the review. Any new sources cited will be added to the list and distinguished from the original sources used.

Sources Used in Current Review

Chen, Y. (Updated 2014 March 23). B-cell leukemia/lymphoma panel. MedlinePlus Medical Encyclopedia [On-line information]. Available online at http://www.nlm.nih.gov/medlineplus/ency/article/003518.htm through http://www.nlm.nih.gov. Accessed December 2014.

(© 1995–2014). Leukemia/Lymphoma Immunophenotyping by Flow Cytometry. Mayo Clinic Mayo Medical Laboratories [On-line information]. Available online at http://www.mayomedicallaboratories.com/test-catalog/Overview/3287 through http://www.mayomedicallaboratories.com. Accessed December 2014.

Maecker, H. et. al. (2012 February 17). Standardizing immunophenotyping for the Human Immunology Project. Nat Rev Immunol v12 (3): 191–200. [On-line information]. Available online at http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3409649/ through http://www.ncbi.nlm.nih.gov. Accessed December 2014.

(Reviewed 2013 July 10). Leukemia – Acute Lymphocytic (Adults). American Cancer Society [On-line information]. Available online at http://www.cancer.org/acs/groups/cid/documents/webcontent/003109-pdf.pdf through http://www.cancer.org. Accessed December 2014.

(2013 December 11). Understanding Laboratory Tests. National Cancer Institute [On-line information]. Available online at http://www.cancer.gov/cancertopics/factsheet/detection/laboratory-tests through http://www.cancer.gov. Accessed December 2014.

(Revised 2012). Understanding Lab and Imaging Tests. Leukemia & Lymphoma Society [On-line information]. Available online through http://www.lls.org. Accessed December 2014.

Sources Used in Previous Reviews

Craig, F. and Foon, K. (2008 April 15). Flow cytometric immunophenotyping for hematologic neoplasms. Blood Journal v111 (8) [On-line information]. Available online at http://bloodjournal.hematologylibrary.org/content/111/8/3941.full through http://bloodjournal.hematologylibrary.org. Accessed April 2011.

Jaffe, E. et. al. (2008 December 1). Classification of lymphoid neoplasms: the microscope as a tool for disease discovery. Blood. 2008 December 1; 112(12): 4384–4399. [On-line information]. Available online at http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2954680/ through http://www.ncbi.nlm.nih.gov. Accessed April 2011.

(Updated 2011 March 13). Blood Tests. Leukemia & Lymphoma Society [On-line information]. Available online at http://www.lls.org/#/diseaseinformation/managingyourcancer/newlydiagnosed/understandingdiagnosis/labimagingtests/bloodtests/ through http://www.lls.org. Accessed April 2011.

Torpy, J. (2009 January 28). Acute Lymphoblastic Leukemia. JAMA Patient Page V301 (4) [On-line information]. PDF available for download at http://jama.ama-assn.org/content/301/4/452.full.pdf through http://jama.ama-assn.org. Accessed April 2011.

Bahler, D. (Updated 2011 February). Lymphoma Phenotyping. ARUP Consult [On-line information]. Available online at http://www.arupconsult.com/Topics/LymphomaPhenotyping.html through http://www.arupconsult.com. Accessed April 2011.

(© 1995–2011). Unit Code 3287: Leukemia/Lymphoma Immunophenotyping by Flow Cytometry. Mayo Clinic, Mayo Medical Laboratory [On-line information]. Available online at http://www.mayomedicallaboratories.com/test-catalog/Overview/3287 through http://www.mayomedicallaboratories.com. Accessed April 2011.

(Reviewed 2010 December). Lymphoid Neoplasms Laboratory Support of Diagnosis and Management Test Guide. Quest Diagnostics [On-line information]. Available online at http://www.questdiagnostics.com/hcp/intguide/jsp/showintguidepage.jsp?fn=TG_Lymphoid_Neoplasms.htm through http://www.questdiagnostics.com. Accessed April 2011.

Wittwera, C. and Brown, M. (2000). Flow Cytometry: Principles and Clinical Applications in Hematology Clinical Chemistry 46:8(B) 1221–1229 [On-line information]. Available online at http://www.clinchem.org/cgi/content/full/46/8/1221 through http://www.clinchem.org. Accessed April 2011.

Mayo Clinic Staff (2010 November 24). Chronic lymphocytic leukemia. MayoClinic [On-line information]. Available online at http://www.mayoclinic.com/health/chronic-lymphocytic-leukemia/DS00565 through http://www.mayoclinic.com. Accessed April 2011.

Acute Leukemia. Merck Manual for Healthcare Professionals [On-line information]. Available online at http://www.merckmanuals.com/professional/sec11/ch142/ch142b.html through http://www.merckmanuals.com. Accessed April 2011.

Pagana, K. D. & Pagana, T. J. (© 2011). Mosby's Diagnostic and Laboratory Test Reference 10th Edition: Mosby, Inc., Saint Louis, MO. Pp 244-247.

Wu, A. (© 2006). Tietz Clinical Guide to Laboratory Tests, 4th Edition: Saunders Elsevier, St. Louis, MO. Pp 1633-1711.

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