Lupus Prone Mice With Defined Mutations

1.6.1. lpr gld and Other CD95 Pathway Mutations This group of recessive mutations share in common defective signaling via the CD95 (Fas)-CD95 ligand apoptosis pathway. The CD95 mutant Ipr arose spontaneously during inbreeding of the MRL Mp strain (100-102) because of a disabling insertion of an endogenous retrotransposon (224-227). The gld mutant arose spontaneously in C3H HeJ mice (228) as a point mutation in CD95 ligand (229,230). Lprcg arose spontaneously in a Japanese CBA KlJms colony...

The Lupus Oriented Mouse Necropsy

Collect urine for analysis (e.g., 248), if desired (see Note 1). a. Cover the bottom of an empty mouse cage with plastic wrap (i.e., no bedding). b. Place mouse on plastic wrap. c. After 10-15 s, remove the mouse. d. Collect urine with a micropipet or Pasteur pipet. 2. Collect blood by retroorbital (RO) approach (e.g., http www.upenn.edu regulatoryaffairs animal annex9.html see Note 2). b. Apply pressure to the external jugular vein caudal to the mandible with thumb and...

John M Walker Series Editor

112 Molecular Cardiology Methods and Protocols, edited by Zhongjie Sun, 2005 111 Chemosensitivity Volume 2, In Vivo Models, Imaging, and Molecular Regulators, edited by Rosalyn D. Blumethal, 2005 110 Chemosensitivity Volume 1, In Vitro Assays, edited by Rosalyn D. Blumethal, 2005 109 Adoptive Immunotherapy, Methods and Protocols, edited by Burkhard Ludewig and Matthias W. Hoffman, 2005 108 Hypertension, Methods and Protocols, edited by J r me P. Fennell and Andrew H. Baker, 2005 107 Human Cell...

Measurement of Cytokines in Autoimmune Disease

Kirou, Christina Lee, and Mary K. Crow Systemic autoimmune diseases are characterized by extensive alterations in immune system function, with cytokines and autoantibodies contributing to impaired immunoregulation and tissue damage. Characterization of the expression and function of cytokines is important for elucidation of pathogenic mechanisms and for identification of therapeutic targets and strategies. We reviewed the utility of assays that reflect individual variability in...

Anti Islet Autoantibody Assays

Type 1A diabetes mellitus, as defined by an expert panel of the American Diabetes Association, is characterized by the presence of antiislet autoantibodies. The presence of islet autoantibodies in individuals who otherwise seem healthy denotes an increased risk for later development of type 1A diabetes. The early expression of insulin autoantibodies (IAAs) is associated with early development of diabetes in young children and NOD mice. IAAs appear early and then disappear at approximately the...

TCell Signaling Abnormalities in Human Systemic Lupus Erythematosus

Nambiar, Sandeep Krishnan, and George C. Tsokos Abnormal expression of key signaling molecules and defective functions of T lymphocytes play a significant role in the pathogenesis of systemic lupus erythematosus SLE . T-cell receptor TCR CD3-mediated stimulation of SLE T cells shows increased protein tyrosine phos-phorylation of cellular proteins, with faster kinetics, heightened calcium response, and decreased interleukin IL -2 production. The molecular mechanism of T-cell...

Pathogenesis and Spectrum of Autoimmunity

The immune system specifically recognizes and eliminates foreign antigens and thus protects the integrity of the host. During maturation of the immune system, tolerance mechanisms develop that prevent or inhibit potentially harmful reactivities to self-antigens. Autoreactive B and T cells that are generated during immune responses are eliminated by apoptosis in the thymus, lymph nodes, or peripheral circulation or are actively suppressed by regulatory T cells. However, autoreactive cells may...

Mitochondrial Hyperpolarization and ATP Depletion Predispose Lupus T Cells to Necrosis

In response to treatment with exogenous H2O2, a precursor, or ROI, lupus T cells failed to undergo apoptosis, and cell death preferentially occurred via necrosis. As noted in ref. 24, H2O2 triggered a rapid increase of AWm and ROI production, which was followed by apoptosis of PBLs in healthy subjects. By contrast, H2O2 failed to elevate AWm, ROI production, and apoptosis, but rather elicited necrosis in patients with lupus. Both CD3 CD28-induced H2O2 production and H2O2-induced apoptosis...

Contents

vii xi 1 Pathogenesis and Spectrum of Autoimmunity Part i. Models of Pathogenesis in Human Autoimmune Disorders 2 Mapping the Systematic Lupus Erythematosus Susceptibility Genes Swapan K. Nath, Jennifer A. Kelly, John B. Harley, 3 T-Cell Signaling Abnormalities in Human Systemic Lupus Erythematosus Madhusoodana P. Nambiar, Sandeep Krishnan, and George C. 4 TCR -Chain Abnormalities in Human Systemic Lupus Erythematosus Madhusoodana P. Nambiar, Sandeep Krishnan, Vishal G. Warke, and George C. 5...

Mitochondrial Checkpoints of TCell Activation and Apoptosis

Ppp Ribose Serine

Both cell proliferation and apoptosis are energy-dependent processes. Energy in the form of adenosine triphosphate ATP is provided through glycolysis and oxidative phosphorylation. The mitochondrion Fig. 1 , the site of oxidative phosphorylation, has long been identified as a source of energy and cell survival 8 . The synthesis of ATP is driven by an electrochemical gradient across the inner mitochondrial membrane maintained by an electron transport chain and the membrane potential negative...

Gradi Frac Control Settings

Control unit UV-1 Knob set to 0.5, switch set to range AU. 2. Pump P-50 Flow rate adjustments will be made using the control panel on the fraction collector. 3. Chart recorder Chart speed set to 5 mm min, blue pen range set to 10 mV, and red pen range set to 1 V. For operation, the valve settings of the GradiFrac are shown in Fig. 1. Fig. 1. Valve settings of the GradiFrac. Fig. 1. Valve settings of the GradiFrac. Fig. 2. Schematic representation of the tubing connection on the main port of...