title: Methods to characterize lymphoid apoptosis in a murine model of autoreactivity.
abstract: The immune system is shaped by the random generation of lymphocytes followed by apoptosis of self-reactive cells, a process termed negative selection. The survival of these pathogenic cells in the periphery can elicit autoreactivity. We describe the development of a biomarker assay for the detection of pathogenic subpopulations of lymphoid cells in adult non-obese diabetic (NOD) mice based on disease-specific alterations in spontaneous or triggered cell death. Utilizing improved methods of cell separations, two distinct lymphoid cell subpopulations with increased susceptibility to apoptosis were identified and quantified. A subpopulation of CD8+ T cells that constitutes approximately 3-7% of the total CD8+ T cell population underwent apoptosis on exposure to low concentrations of TNF-alpha. Such cells were exclusively detected only in NOD mice with histologic signs of active autoreactivity. The non-T cell compartment of NOD immune system, although resistant to TNF-alpha-induced apoptosis, contained a subpopulation of B cells with spontaneous death by culture alone. The refined detection of small numbers of lymphoid cell subsets with quantifiable differences in apoptosis provides a possible immune biomarker for monitoring disease activity or treatment interventions.
authors.affiliation: Immunobiology Laboratory, Massachusetts General Hospital and Harvard Medical School, Building 149, 13th Street, CNY-3601, Charlestown, MA 02129, USA.
authors.forename: Willem M
authors.initials: WM
authors.lastname: Kühtreiber
authors.forename: Shohta
authors.initials: S
authors.lastname: Kodama
authors.forename: Douglas E
authors.initials: DE
authors.lastname: Burger
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Dale
authors.forename: Denise L
authors.initials: DL
authors.lastname: Faustman
mesh_terms: D000818:Animals; D017209:Apoptosis; D015415:Biomarkers / Q000032:analysis; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology* / Q000473:pathology; D005260:Female; D016131:Lymphocyte Subsets / Q000276:immunology*; D051379:Mice; D016688:Mice, Inbred NOD; D013154:Spleen / Q000187:drug effects / Q000276:immunology / Q000473:pathology; D016176:T-Lymphocyte Subsets / Q000276:immunology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D015415:Biomarkers; D014409:Tumor Necrosis Factor-alpha