CLINICAL FLOW CYTOMETRIC ANALYSIS


Director:  Giovanni Insuasti-Beltran, M.D. (501-686-6527); pager (501-405-6934)
Manager: Curtis Wade, BS, MLS(ASCP)CM (501-686-5226)
Supervisor:  Linda Whigham, AIBMS (501-686-5953)
Technical Specialist: Rebecca Owens, BSMT (501-686-5953)
Hours: Monday through Friday, 8:00am - 5:00pm


Flow Cytometric Analysis of Cell Surface Antigens (Immunophenotype)

Clinical Significance:
Changes seen in lymphocyte subsets, such as T or B cell populations, can be indicative of immunological changes produced by various diseases or treatments.  The Helper/Suppressor Panel is used as a general screen for all lymphocyte subsets in cases where there is no knowledge of the underlying cause or disease which may result in the patient's health problems.  Acquired Immune Deficiency Syndrome (AIDS) has several clinical symptoms, one of which is an abnormal decrease in the T helper cell population or CD4 in the peripheral blood.  This abnormality can show up early in the disease and can be followed during the course of the disease and during various treatment programs as a monitor of the progression of the disease or the effectiveness of the treatment.  After a bone marrow transplant, lymphocyte subsets are monitored using the Immune Studies panel to determine the status of certain relevant subsets.

Organ and bone marrow transplants have fast become a well established treatment for patients with chronic life threatening diseases.  Flow cytometry has proven to be a valuable aid in evaluating these patients for disease remission, transplant engraftment or rejection and for response to growth factors (CD34) or immunotherapy (CD3).


The diagnoses of leukemia and lymphoma have, until recently, consisted of evaluating the specimen by various morphological and histochemical criteria.  More extensive immunophenotyping panels by flow cytometry are used to distinguish Chronic Lymphocytic Leukemia from a generalized increase of lymphocytes (Chronic Leukemia/Lymphoma Panel) and to subclassify different chronic leukemias. Flow Cytometry is also used to distinguish Acute Lymphoid Leukemia from Acute Myeloid Leukemia (Acute Leukemia Panel).

Flow cytometry is not indicated for use in the diagnosis of Chronic Myeloid Leukemia in the chronic stage.  Flow cytometry can also only be used to rule out Non-Hodgkin's Lymphoma but not to diagnose Hodgkin's Lymphoma.

Cylex is a non-flow cytometric assay performed in the Flow Cytometry lab.  Cylex, an immune function assay that detects cell-mediated immunity status of CD4 cells, can be used as an additional test for monitoring immunosuppressed patients.  This test can be used in conjunction with CD4 enumeration or in place of the CD4 count.  Discussion of this test and specimen requirements for it are discussed at the end of the discussion of Flow Cytometry assays.

I.    Lymphocyte Subset Analysis for evaluating immunological status.

II.    Chronic Leukemia/Lymphoma and Acute Leukemia Phenotype

        The following tests do not have SoftLab results.  These Flow Cytometry tests should be ordered by panel
        name either in Sunrise or on the requisition sheet.  Results can be found in SOFTPath.

III.    Transplantation Panels:


General Specimen Information

Specimen Requirements for Flow Cytometry Tests: 

          Peripheral Blood for  CD4, Immune Studies, Helper/Suppressor Panels, and CD34
         
1 EDTA (Lavendar Top)

Peripheral Blood (for all other tests)
1 EDTA tube (Lavender Top)

Bone Marrow Aspirate
Orders for flow cytometry tests should be made when arrangements are made for the bone marrow procedure.

Biopsies and Solid Tissues
Orders for flow cytometry tests should be made when arrangements are made for the biopsy or surgical procedure through the Department of Pathology or Radiology.

Body Fluids
All body fluids should be obtained and held at room temperature before being transported to the laboratory.  The specimens should be received in the laboratory within 24 hours of being obtained from the patient.
 

Specimen Rejection:

  1. Clotted tubes
  2. Collected in improper tube
  3. Partial draw
  4. Insufficient number of viable cells
  5. Specimen older than 72 hours
     

Hours of Operation:
Samples are accepted through the Clinical Laboratory Specimen Receiving Area at all hours.  All specimens must be received within 24 hours of collection.  The regular work hours for Flow Cytometry are 7:30am to 5:30pm Monday through Friday.  Any specimen received outside of these hours will be held until the next regular work day.  Contact the Flow Cytometry lab during normal working hours at 501-686-5953 for additional instructions for after-hour specimens.


Results Turnaround Time



Table of Tests Performed in Flow Cytometry


Test Name
(TEST CODE)

Specimen Requirements
 

Therapeutic Range

Expected
Turnaround
Time

CD3 Lymphocyte Levels
(CD3/5)
1 Lav. top
 
Normal ranges:
CD3:  68-80%; 1117-2250/Ál
CD5:  70-82%; 700-3280/Ál
Therapeutic range CD3 or CD5 =
<
10%;  < 100/Ál
4-24 hrs
CD4 T Cell Subset
(CD4)
1 Lav top 39-57%, 720-1348/Ál 4-24 hrs
CD34 [Human Progenitor Cell Antigen
(APB34) {ProCount}
1 Lav top Normal range = Negative
Therapeutic range is any increase over the normal range.
4-24 hrs
Helper/Suppressor Cell Panel
(H/S)
1 Lav top CD3:  64-82%, 1171-2005/Ál
CD4:  39-57%, 720-1348/Ál
CD8:  17-31%, 318-710/Ál
CD19:  8-16%, 151-343/Ál
CD16/56:  7-21, 145-453/Ál
H/S Ratio 1.0-3.6
4-24 hrs
Immune Studies Panel
(IMSPB)
1 Lav top CD3:  64-82%, 1171-2005/Ál
CD4:  39-57%, 720-1348/Ál
CD19:  8-16%, 151-343/Ál
4-24 hrs
Chronic Leukemia/Lymphoma Panel 1 Lav top, Bone marrow aspirate, Biopsy, Tissue or Body Fluid Interpretive report.  Includes enumeration of all lymphocytes and subsets. 24-48 hrs
Sezary Cell/Mycosis Fungoides Panel 1 Lav top, Bone marrow aspirate, Biopsy, Tissue or Body Fluid Interpretive report.  Includes enumeration of all lymphocytes and subsets. 24-48 hrs
Acute Leukemia Panel 1 Lav top, Bone marrow aspirate, Biopsy, Tissue or Body Fluid Interpretive report.  Includes enumeration of blasts with T cell, B cell and myelo/monocytic cell markers 24-48 hrs
Cytoplasmic Immunoglobulins (CIg) vs DNA 1 Green Sodium Heparin top + 1 Lav top, Bone marrow aspirate, Biopsy, Tissue or Body Fluid Interpretive report. 24-48 hrs
Plasma Cell Phenotype
 
1 Lav top, Bone marrow aspirate, Biopsy, Tissue or Body Fluid Interpretive report. 24-48 hrs

Updated 6.28.2013

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