T-Cell Subsets, Naive, Memory and Activated
Clinical Information Discusses physiology, pathophysiology, and general clinical aspects, as they relate to a laboratory test
T cells, after completing development and initial differentiation in the thymus, enter the periphery as naive (n) T cells. Naive T cells undergo further differentiation into effector and memory T cells in the peripheral lymphoid organs after recognizing specific antigenic peptides in the context of major histocompatibility (MHC) molecules, through the antigen-specific T-cell receptor. In addition to the cognate signal of the peptide-MHC complex interaction (the term cognate refers to 2 biological molecules that normally interact), T cells require additional costimulatory signals to complete T-cell activation. Naive T cells circulate continuously through the lymph nodes and, on recognition of specific antigen, undergo activation. Due to their antigen-inexperienced state, naive T cells require activation by more potent antigen-presenting cells, such as dendritic cells.
Naive T cells can survive in circulation for prolonged periods of time and are very important in contributing to T-cell repertoire diversity. They proliferate in response to interleukin-2, as a consequence of their response to antigen through recognition of peptide-MHC costimulation. These expanded antigen-specific T cells undergo further differentiation into effector cells. The differentiation of naive CD8 T cells into cytotoxic effectors capable of killing target T cells loaded with endogenous peptides on MHC class I molecules may require additional costimulatory signals from CD4 T cells. Naive CD4 T cells also differentiate into different effector subsets such as Th1, Th2, and Th17, which produce specific cytokines.(1)
T cells can be subdivided into naive and memory subsets based on the expression of cell-surface markers, such as CD45RA and CD45RO, among others. It was initially thought that the presence of cell-surface CD45RA indicated the naive subset, while the presence of CD45RO indicated memory subsets. But, it has now been shown that multiple, rather than single, markers are required to distinguish these subsets.(2) Lanzavecchia and Sallusto proposed a model where naive T cells expressing CD45RA and CCR7 lose CD45RA expression on recognition of antigen.(3) The surface markers for identifying naive T-cell subsets include CD45RA, CD62L (L-selectin), and CD27.(4,5)
Memory T cells are antigen-experienced cells that are present in greater numbers than antigen-specific precursors, and can respond more efficiently and rapidly to specific antigen. Memory T cells can maintain their populations independent of antigen by homeostatic proliferation in response to cytokines. While there are subcategories of memory T cells based on effector function and cell surface and cytolytic molecule expression, the 2 main categories of memory T cells are central memory T cells (Tcm) and effector memory T cells (Tem).(1,6)
Tcm express the CD45RO molecule along with CD62L (L-selectin) and CCR7, and are present mainly in lymphoid tissue.(6,7) They are able to respond to antigen through rapid proliferation and expansion and differentiation into Tem. By themselves, Tcm are not directly effective in effector cytolytic function.
Unlike Tcm, Tem express only CD45RO (not CD62L and CCR7).(6) As the name suggests, Tem have remarkable effector function, though they do not proliferate well. Tem are present throughout the circulation in peripheral tissues providing immune surveillance.
Memory T cells are particularly important for maintenance of immune competence since they are associated with a rapid and effective response to pathogens. Therefore, depletion of this compartment has more immediate significance than the depletion of naive T cells.
Activation of human T cells is critical for the optimal and appropriate performance of T-cell functions within the adaptive immune response. Activated naive T cells undergo proliferation, as well as subsequent differentiation into effector T cells, and are capable of producing cytokines that can modulate the immune response in a variety of ways.(8) There are several markers associated with T-cell activation, but those most commonly used include CD25 (IL-25R)(8) and MHC class II.(9) Additionally, the expression of the costimulatory molecule CD28 augments the T-cell activation response.(10)
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 am and noon, with no change between noon and afternoon. Natural killer cell counts, on the other hand, are constant throughout the day.(11) Circadian variations in circulating T-cell counts have been shown to be negatively correlated with plasma cortisol concentration.(12-14) In fact, cortisol and catecholamine concentrations control distribution and, therefore, numbers of naive versus effector CD4 and CD8 T cells.(11) It is generally accepted that lower CD4 T-cell counts are seen in the morning compared with the evening(15), and during summer compared to winter.(16) These data, therefore, indicate that timing and consistency in timing of blood collection is critical when serially monitoring patients for lymphocyte subsets.
Determining the presence of naive, memory, and activated T cells in various clinical contexts including autoimmune diseases, immunodeficiency states, T-cell recovery posthematopoietic stem cell transplant, DiGeorge syndrome, and as a measure for T-cell immune competence
Naive T-cells results can be used as a surrogate marker for thymic-derived T-cell reconstitution, when used in conjunction with assessment of T-cell receptor excision circles (TREC / T-Cell Receptor Excision Circles [TREC] Analysis for Immune Reconstitution)
Assessing a patient's relative risk for infections
Evaluation of patients with cellular or combined primary immunodeficiencies
Evaluation of T-cell reconstitution after hematopoietic stem cell transplant, chemotherapy, biological therapy, immunosuppression or immunomodulator therapy
Evaluation of patients with autoimmune diseases
Evaluation of HIV-positive patients for naive and memory subsets
Evaluation of T-cell immune competence (presence of memory and activated T cells) in patients with recurrent infections
Absence or reduction of naive T cells with or without T-cell lymphopenia indicates absent or impaired T-cell reconstitution or thymic output. Reduction in activated T cells can also indicate a reduced T-cell immune competent state.
Increases in naive T cells with corresponding decreases in the memory T-cell compartment indicates a failure of further differentiation and effector function or selective loss of memory T cells and an increased risk for infection.
Cautions Discusses conditions that may cause diagnostic confusion, including improper specimen collection and handling, inappropriate test selection, and interfering substances
This assay provides quantitative information on various T-cell subsets in blood; it does not provide any information on the antigen-specific or otherwise functional state of the T cells. To assess the overall functional state of T cells, LPMGF / Lymphocyte Proliferation to Mitogens, Blood and LPAGF / Lymphocyte Proliferation to Antigens, Blood (using Candida and tetanus antigens) are appropriate. To assess cytomegalovirus (CMV)-specific immune competence, order CMVC8 / Cytomegalovirus (CMV) CD8 T-Cell Immune Competence, Quantitative Assessment by Flow Cytometry.
Timing and consistency in timing of blood collection is critical when serially monitoring patients for lymphocyte subsets. See data under Clinical Information.
Reference Values Describes reference intervals and additional information for interpretation of test results. May include intervals based on age and sex when appropriate. Intervals are Mayo-derived, unless otherwise designated. If an interpretive report is provided, the reference value field will state this.
The appropriate age-related reference values will be provided on the report.
Clinical References Provides recommendations for further in-depth reading of a clinical nature
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2. De Rosa SC, Herzenberg LA, Herzenberg LA, et al: 11-color, 13-parameter flow cytometry: identification of human naive T-cells by phenotype, function, and T-cell receptor diversity. Nat Med 2001;7:245-248
3. Sallusto F, Lenig D, Forster R, et al: Two subsets of memory T-lymphocytes with distinct homing potentials and effector functions. Nature 1999;401:708-712
4. Picker LJ, Treer JR, Ferguson-Darnell B, et al: Control of lymphocyte recirculation in man. I. Differential regulation of the peripheral lymph node homing receptor L-selectin on T-cells during the virgin to memory cell transition. J Immunol 1993;150:1105-1121
5. Morimoto C, Schlossman SF: Human naive and memory T-cells revisited: new markers (CD31 and CD27) that help define CD4+ T-cell subsets. Clin Exp Rheumatol 1993;11:241-247
6. LaRosa DF, Orange JS: Lymphocytes. J Allergy Clin Immunol 2008;121:S364-369
7. Foster AE, Marangolo M, Sartor MM, et al: Human CD62L-memory T-cells are less responsive to alloantigen stimulation than CD62L+ naive T-cells: potential for adoptive immunotherapy and allodepletion. Blood 2004;104:2403-2409
8. Brenchley JM, Douek DC, Ambrozal DR, et al: Expansion of activated human naive T-cells preceded effector function. Clin Exp Immunol 2002;130:431-440
9. Holling TM, van der Stoep N, Quinten E, et al: Activated human T-cells accomplish MHC class II expression through T-cell specific occupation of class II transactivator promoter III. J Immunol 2002;168:763-770
10. Thompson CB, Lindsten T, Ledbetter JA, et al: CD28 activation pathway regulates the production of multiple T-cell-derived lymphokines/cytokines. Proc Natl Acad Sci USA 1989;86:1333-1337
11. Carmichael KF, Abayomi A: Analysis of diurnal variation of lymphocyte subsets in healthy subjects and its implication in HIV monitoring and treatment. 15th Intl Conference on AIDS, Bangkok, Thailand, 2004, Abstract B11052
12. Dimitrov S, Benedict C, Heutling D, et al: Cortisol and epinephrine control opposing circadian rhythms in T cell subsets. Blood 2009;113(21):5134-5143
13. Dimitrov S, Lange T, Nohroudi K, Born J: Number and function of circulating antigen presenting cells regulated by sleep. Sleep 2007;30:401-411
14. Kronfol Z, Nair M, Zhang Q, et al: Circadian immune measures in healthy volunteers: relationship to hypothalamic-pituitary-adrenal axis hormones and sympathetic neurotransmitters. Psychosom Med 1997;59:42-50
15. Malone JL, Simms TE, Gray GC, et al: Sources of variability in repeated T-helper lymphocyte counts from HIV 1-infected patients: total lymphocyte count fluctuations and diurnal cycle are important. J AIDS 1990;3:144-151
16. Paglieroni TG, Holland PV: Circannual variation in lymphocyte subsets, revisited. Transfusion 1994;34:512-516