2009The Journal of ImmunologyRequires access

Age-dependent T and B lymphocyte subsets defined by multicolor flow cytometry (86.13)

Matthew Crabtree, Stephen C. DeRosa, Gordon Alan Cohen, ALICE E. WIEDEMAN, Anne M. Stevens

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Abstract

Abstract Immune dysregulation in children due to primary immunodeficiency, autoimmune disease or immunomosuppressive therapy may be characterized in detail by multiparametric flow cytometry. To determine age-dependent reference values for healthy children, we developed four panels: aß and γδ T lymphocytes; regulatory T lymphocytes; B lymphocytes; NK and NK T cells. Detailed analyses performed on 18 healthy subjects aged 8 to 46 years revealed differences between adult and juvenile phenotypes in 66 of 301 distinct subsets. Among naïve aß T lymphocytes (CD45RA+CCR7+CD27+) we found a 60% decrease in the CD4+ subset but no change in the CD8+ subset in adults compared to children. Expression of CD31, an aß T lymphocyte marker of recent thymic emigration, decreased 50% with age. T regulatory cells (CD4+CD127loCD25+FoxP3+) were 30% lower in adults. No differences were found in major γδ T lymphocyte, NK, or NK T cell subsets. Memory B cells (CD38loCD24+) increased 2-fold with age, while naïve B cells (CD24hiCD38+IgM+IgD-) decreased by 50%. Thus, the influence of aging greatly effects subset frequencies, and may reflect decreasing thymus function and acute/chronic stimulation of memory B and T lymphocytes. Phenotypic analysis of leukocyte subsets may serve as a valuable tool for detecting and monitoring immune dysregulation in children, but age-matched controls are essential. Supported by the McMillen Foundation

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Abstract Immune dysregulation in children due to primary immunodeficiency, autoimmune disease or immunomosuppressive therapy may be characterized in detail by multiparametric flow cytometry. To determine age-dependent reference values for healthy children, we developed four panels: aß and γδ T lymphocytes; regulatory T lymphocytes; B lymphocytes; NK and NK T cells. Detailed analyses performed on 18 healthy subjects aged 8 to 46 years revealed differences between adult and juvenile phenotypes in 66 of 301 distinct subsets. Among naïve aß T lymphocytes (CD45RA+CCR7+CD27+) we found a 60% decrease in the CD4+ subset but no change in the CD8+ subset in adults compared to children. Expression of CD31, an aß T lymphocyte marker of recent thymic emigration, decreased 50% with age. T regulatory cells (CD4+CD127loCD25+FoxP3+) were 30% lower in adults. No differences were found in major γδ T lymphocyte, NK, or NK T cell subsets. Memory B cells (CD38loCD24+) increased 2-fold with age, while naïve B cells (CD24hiCD38+IgM+IgD-) decreased by 50%. Thus, the influence of aging greatly effects subset frequencies, and may reflect decreasing thymus function and acute/chronic stimulation of memory B and T lymphocytes. Phenotypic analysis of leukocyte subsets may serve as a valuable tool for detecting and monitoring immune dysregulation in children, but age-matched controls are essential. Supported by the McMillen Foundation

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Available abstract

Abstract Immune dysregulation in children due to primary immunodeficiency, autoimmune disease or immunomosuppressive therapy may be characterized in detail by multiparametric flow cytometry. To determine age-dependent reference values for healthy children, we developed four panels: aß and γδ T lymphocytes; regulatory T lymphocytes; B lymphocytes; NK and NK T cells. Detailed analyses performed on 18 healthy subjects aged 8 to 46 years revealed differences between adult and juvenile phenotypes in 66 of 301 distinct subsets. Among naïve aß T lymphocytes (CD45RA+CCR7+CD27+) we found a 60% decrease in the CD4+ subset but no change in the CD8+ subset in adults compared to children. Expression of CD31, an aß T lymphocyte marker of recent thymic emigration, decreased 50% with age. T regulatory cells (CD4+CD127loCD25+FoxP3+) were 30% lower in adults. No differences were found in major γδ T lymphocyte, NK, or NK T cell subsets. Memory B cells (CD38loCD24+) increased 2-fold with age, while naïve B cells (CD24hiCD38+IgM+IgD-) decreased by 50%. Thus, the influence of aging greatly effects subset frequencies, and may reflect decreasing thymus function and acute/chronic stimulation of memory B and T lymphocytes. Phenotypic analysis of leukocyte subsets may serve as a valuable tool for detecting and monitoring immune dysregulation in children, but age-matched controls are essential. Supported by the McMillen Foundation

Key concepts: Immunology, Immune system, CD8, FOXP3, Flow cytometry, Immunophenotyping, Biology, Immune dysregulation

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