% CD3+CD25+ Lymphs

Optimal Result: 4.9 - 25.9 %.

CD25 is the receptor for IL2 and is expressed on activated T cells, B cells, and macrophages. CD25 is expressed in certain types of B-cell lymphoma (hairy cell leukemia) and T-cell lymphoma (adult T-cell lymphoma/leukemia [ATLL]).

The interleukin-2 receptor is designated CD25. Originally isolated from T-lymphocytes, it is now known to be expressed on hairy cell leukemia and adult T-cell leukemia/lymphoma, classical Hodgkin lymphoma, and a subset of other peripheral T-cell lymphomas.

General information:

Lymphocytes in peripheral blood (circulation) are heterogeneous and can be broadly classified into T cells, B cells, and natural killer (NK) cells.

There are various subsets of each of these individual populations with specific cell-surface markers and function.

This assay provides absolute (cells/mcL) and relative (%) quantitation for the main categories of T cells, B cells, and NK cells, in addition to a total lymphocyte count (= CD45+).

- Each of these lymphocyte subpopulations have distinct effector and regulatory functions and are maintained in homeostasis under normal physiological conditions.

- Each of these lymphocyte subsets can be identified by a combination of one or more cell surface markers.


The CD3 antigen is a pan-T-cell marker, and T cells can be further divided into 2 broad categories, based on the expression of CD4 or CD8 coreceptors.

B cells:

B cells can be identified by expression of CD19.

NK cells: 

NK cells are typically identified by the coexpression of CD16 and CD56.


Total T-cells (CD3+)

B-cells (CD3-/CD19+)

Natural killer (NK) cells (CD3-/CD56+CD16+)

Helper/inducer (CD3+/CD4+) T-cell subset

cytotoxic/suppressor (CD3+/CD8+) T-cell subset

Total CD45+ lymphocyte count

CD4:CD8 ratio


The absolute counts of lymphocyte subsets are known to be influenced by a variety of biological factors, including hormones, the environment, and temperature. The studies on diurnal (circadian) variation in lymphocyte counts have demonstrated progressive increase in CD4 T-cell count throughout the day, while CD8 T cells and CD19+ B cells increase between 8:30 a.m. and noon with no change between noon and afternoon. NK-cell counts, on the other hand, are constant throughout the day.

Circadian variations in circulating T-cell counts have been shown to be negatively correlated with plasma cortisol concentration. In fact, cortisol and catecholamine concentrations control distribution and, therefore, numbers of naive versus effector CD4 and CD8 T cells.

It is generally accepted that lower CD4 T-cell counts are seen in the morning compared to the evening and during summer compared to winter. These data, therefore, indicate that timing and consistency in timing of blood collection is critical when serially monitoring patients for lymphocyte subsets.

Abnormalities in the number and percent of T (CD3, CD4, CD8), B (CD19), and NK (CD16+CD56) lymphocytes have been described in a number of different disease conditions.

In patients who are infected with HIV, the CD4 count is measured for 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 patients who are HIV-positive be tested every 3 to 6 months for the level of CD4 T lymphocytes.

Lymphocyte subset quantitation is also very useful in the evaluation of patients with primary immunodeficiencies of all ages, including follow-up for newborn screening for severe combined immunodeficiency and immune monitoring following immunosuppressive therapy for transplantation, autoimmunity, or any other relevant clinical condition where immunomodulatory treatment is used.

It is also helpful as a preliminary screening assay for gross quantitative anomalies in any lymphocyte subset, whether related to malignancies or infection.

The 2008 guidelines for diagnosis and treatment of Chronic Lymphocytic Leukemia (CLL) from the International Workshop on Chronic Lymphocytic Leukemia recommends changing the diagnostic criteria for CLL from an absolute lymphocyte count greater than 5 x 10(9)/L to a circulating B-cell count greater than 5 x 10(9)/L(8,9) previously defined in the 1996 National Cancer Institute (NCI) guidelines for CLL. This flow cytometric assay enables accurate quantitation of circulating B cells using a single platform technology with absolute quantitation through the use of flow cytometry beads.






Milioglou I, Kalaitzidou I, Ladomenou F. Interpretation of lymphocyte subset counts by the general pediatrician. Pediatr Int. 2019 Jan;61(1):16-22. doi: 10.1111/ped.13701. Epub 2018 Dec 10. PMID: 30248214.

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