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Unit Code 9336:
T- and B-Cell Quantitation by Flow Cytometry

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Useful For

Monitoring CD4 counts and assessing immune deficiencies

Clinical Information

Normal immunity requires a balance between the activities of various

lymphocyte subpopulations with different effector and regulatory functions.

 

Different immune cells can be characterized by unique surface membrane

antigens described by a cluster of differentiation nomenclature (eg, CD3

is an antigen found on the surface of T-lymphocytes). Abnormalities in the

number and percent of T (CD3), T-helper (CD4), T-suppressor (CD8),

B (CD19), and natural killer (CD16 CD56) lymphocytes have been

described in a number of different diseases. In patients who are infected

with human immunodeficiency virus (HIV), the CD4 count is measured for

acquired immune deficiency syndrome (AIDS) diagnosis and for initiation

of antiviral therapy. The progressive loss of CD4 T-lymphocytes in patients

infected with HIV is associated with increased infections and complications.

 

The Public Health Service has recommended that all HIV-positive patients

be tested every 3 to 6 months for the level of CD4 T-lymphocytes.

Reference Values

T & B SURFACE MARKER

      % T-cells (CD3)

                0-2 months: 53-84%*

                3-5 months: 51-77%*

                6-11 months: 49-76%*

                12-23 months: 53-75%*

                2-5 years: 56-75%*

                6-11 years: 60-76%*

                12-17 years: 56-84%*

                >=18 years: 52-84%

      % B-cells (CD19)

                0-2 months: 6-32%*

                3-5 months: 11-41%*

                6-11 months: 14-37%*

                12-23 months: 16-35%*

                2-5 years: 14-33%*

                6-11 years: 13-27%*

                12-17 years: 6-23%*

                >=18 years: 5-25%

      % Natural killer (CD16 CD56)

                0-2 months: 4-18%*

                3-5 months: 3-14%*

                6-23 months: 3-15%*

                2-11 years: 4-17%*

                12-17 years: 3-22%*

                >=18 years: 5-30%

      % Helper cells (CD4)

                0-2 months: 35-64%*

                3-5 months: 35-56%*

                6-11 months: 31-56%*

                12-23 months: 32-51%*

                2-5 years: 28-47%*

                6-11 years: 31-47%*

                12-17 years: 31-52%*

                >=18 years: 30-61%

      % Suppressor cells (CD8)

                0-2 months: 12-28%*

                3-5 months: 12-23%*

                6-11 months: 12-24%*

                12-23 months: 14-30%*

                2-5 years: 16-30%*

                6-17 years: 18-35%*

                >=18 years: 12-42%

ABSOLUTE COUNTS

      CD45 lymph count, flow:  0.66-4.60 thou/uL

      T-cells (CD3)

                0-2 months: 2,500-5,500 cells/uL*

                3-5 months: 2,500-5,600 cells/uL*

                6-11 months: 1,900-5,900 cells/uL*

                12-23 months: 2,100-6,200 cells/uL*

                2-5 years: 1,400-3,700 cells/uL*

                6-11 years: 1,200-2,600 cells/uL*

                12-17 years: 1,000-2,200 cells/uL*

                > or =18 years: 582-1,992 cells/uL

      B-cells (CD19)

                0-2 months: 300-2,000 cells/uL*

                3-5 months: 430-3,000 cells/uL*

                6-11 months: 610-2,600 cells/uL*

                12-23 months: 720-2,600 cells/uL*

                2-5 years: 390-1,400 cells/uL*

                6-11 years: 270-860 cells/uL*

                12-17 years: 110-570 cells/uL*

                > or =18 years: 71-567 cells/uL

      Natural killer (CD16 CD56)

                0-2 months: 170-1,100 cells/uL*

                3-5 months: 170-830 cells/uL*

                6-11 months: 160-950 cells/uL*

                12-23 months: 180-920 cells/uL*

                2-5 years: 130-720 cells/uL*

                6-11 years: 100-480 cells/uL*

                12-17 years: 70-480 cells/uL*

                > or =18 years: 80-597 cells/uL

      Helper cells (CD4)  

                0-2 months: 1,600-4,000 cells/uL*

                3-5 months: 1,800-4,000 cells/uL*

                6-11 months: 1,400-4,300 cells/uL*

                12-23 months: 1,300-3,400 cells/uL*

                2-5 years: 700-2,200 cells/uL*

                6-11 years: 650-1,500 cells/uL*

                12-17 years: 530-1,300 cells/uL*

                > or =18 years: 401-1,532 cells/uL

      Suppressor cells (CD8)

                0-2 months: 560-1,700 cells/uL*

                3-5 months: 590-1,600 cells/uL*

                6-11 months: 500-1,700 cells/uL*

                12-23 months: 620-2,000 cells/uL*

                2-5 years: 490-1,300 cells/uL*

                6-11 years: 370-1,100 cells/uL*

                12-17 years: 330-920 cells/uL*

                > or =18 years: 152-838 cells/uL

LYMPHOCYTE RATIO

      H/S ratio:  > or = 1.0

*Shearer WT, Rosenblatt HM, Gelman RS, et al:  Lymphocyte

subsets in healthy children from birth through 18 years of age: The

Pediatric AIDS Clinical Trials Group P1009 study. J Allergy Clin Immunol   

2003;112(5):973-980

Interpretation

When the CD4 count falls below 500 cell/uL, HIV-positive patients

can be diagnosed with AIDS and can receive antiretroviral therapy.

 

When the CD4 count falls below 200 cells/uL, prophylaxis against

Pneumocystis jiroveci pneumonia is recommended.

Cautions

This assay is not used for diagnosing lymphocytic malignancies or

evaluation of lymphocytosis of unknown etiology. In these situations,

#3287 "Leukemia Immunophenotyping by Flow Cytometry" which

includes a hematopathology review, should be ordered.

Clinical Reference

1.   Mandy FF, Nicholson JK, McDougal JS: Guidelines for performing

      single-platform absolute CD4 T-cell determinations with CD45 gating

      for persons infected with human immunodeficiency virus. Center for

      Disease Control and Prevention. MMWR Morb Mortal Wkly Rep

      2003;52:1-13                

 

2.   Centers for Disease Control: 1997 Revised guidelines for performing

      CD4 T-cell determinations in persons infected with human

      immunodeficiency virus (HIV). MMWR Morb Mortal Wkly Rep 46 no.

      RR-2: 1997, pp 1-29

 

3.   U.S. Department of Health and Human Services:  Recommendations

      for prophylaxis against Pneumocystis carinii pneumonia for adults

      and adolescents infected with human immunodeficiency virus.

      MMWR Morb Mortal Wkly Rep 43 no. RR-3: 1994, pp 1-21

Key