Thrombospondin and tumor necrosis factor

Progress in two main projects in my laboratory are discussed: (i) the study of the extracellular matrix/cell surface molecule thrombospondin (TSP); and (ii) the characterization of primary or immediate early response genes induced by tumor necrosis factor-α (TNF)

Corrigendum to: Activation of caspases triggered by cytochrome c in vitro (FEBS 20097)

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/117119/1/feb2s0014579398004657.pd

Expression of thrombospondin in the adult nervous system

Thrombospondin (TSP) is an extracellular matrix molecule that has been previously associated with neural development and neurite outgrowth in vitro. Little is known, however, about the expression of TSP in the adult nervous system. In this study, TSP localization was examined in nervous tissue from adult mouse, goldfish, newt, and adult and juvenile Xenopus. TSP was associated with neurons in the brains of all species examined. TSP was present in central nerve tracts capable of regeneration, such as the goldfish, Xenopus, and newt optic nerves, but was absent from tracts not capable of regeneration, such as the mouse optic nerve. TSP was also present in the neuropil of goldfish and newt spinal cord, but was restricted to motor neurons in mice and adult Xenopus. In addition, TSP was observed in sciatic nerves of mice, Xenopus, and newt. These results indicate a correlation between the presence of TSP and the potential for successful nerve regeneration across a wide range of animal classes. © Wiley-Liss, Inc.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/50059/1/903400109_ftp.pd

FADD, a novel death domain-containing protein, interacts with the death domain of fas and initiates apoptosis

AbstractUsing the cytoplasmic domain of Fas in the yeast two-hybrid system, we have identified a novel interacting protein, FADD, which binds Fas and Fas-FD5, a mutant of Fas possessing enhanced killing activity, but not the functionally inactive mutants Fas-LPR and Fas-FD8. FADD contains a death domain homologous to the death domains of Fas and TNFR-1. A point mutation in FADD, analogous to the Ipr mutation of Fas, abolishes its ability to bind Fas, suggesting a death domain to death domain interaction. Overexpression of FADD in MCF7 and BJAB cells induces apoptosis, which, like Fas-induced apoptosis, is blocked by CrmA, a specific inhibitor of the interieukin-lβ-converting enzyme. These findings suggest that FADD may play an important role in the proximal signal transduction of Fas

Innate immunity against Francisella tularensis is dependent on the ASC/caspase-1 axis

Francisella tularensis is a highly infectious gram-negative coccobacillus that causes the zoonosis tularemia. This bacterial pathogen causes a plague-like disease in humans after exposure to as few as 10 cells. Many of the mechanisms by which the innate immune system fights Francisella are unknown. Here we show that wild-type Francisella, which reach the cytosol, but not Francisella mutants that remain localized to the vacuole, induced a host defense response in macrophages, which is dependent on caspase-1 and the death-fold containing adaptor protein ASC. Caspase-1 and ASC signaling resulted in host cell death and the release of the proinflammatory cytokines interleukin (IL)-1β and IL-18. F. tularensis–infected caspase-1– and ASC-deficient mice showed markedly increased bacterial burdens and mortality as compared with wild-type mice, demonstrating a key role for caspase-1 and ASC in innate defense against infection by this pathogen

Inhibitory Effect of Gamma Interferon on Cultured Human Keratinocyte Thrombospondin Production, Distribution, and Biologic Activities

Rapidly proliferating keratinocytes (KCs) maintained in low calcium, serum-free medium produce and utilize thrombospondin (TSP) as an attachment and spreading factor. To begin to understand the modulation of KC TSP metabolism, gamma interferon (IFN-γ), a product of activated T lymphocytes, was added to KC cultures. IFN-γ; was chosen because activated T cells appear at sites of cutaneous injury. Two additional cytokines including tumor necrosis factor (TNF) and IFN-β were also examined. IFN-γ (600 U/ml), but not TNF (500 U/ml) or IFN-β (103 U/ml), as single agents decreased KC TSP biosynthesis, secretion, and utilization as an attachment factor. IFN-γ alone did not detectably decrease TSP mRNA levels suggesting a post-transcriptional effect in KCs. However, the combination of IFN-γ (600 U/ml) and TNF (500 U/ml) inhibited TSP mRNA production. These results demonstrate the modulation of KC TSP metabolism and biologic activity

TRUNDD, a new member of the TRAIL receptor family that antagonizes TRAIL signalling

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/116376/1/feb2s0014579398001355.pd

A Role for FADD in T Cell Activation and Development

AbstractFADD is a cytoplasmic adapter molecule that links the family of death receptors to the activation of caspases during apoptosis. We have produced transgenic mice expressing a dominantly interfering mutant of FADD, lacking the caspase-dimerizing death effector domain, as well as mice overexpressing the poxvirus serpin, CrmA, an inhibitor of caspases downstream of FADD. While thymocytes from either line of mice were completely protected from CD95-dependent cytotoxicity, neither transgene afforded protection from apoptosis induced during thymocyte selection and neither led to the lymphoproliferative disorders associated with deficiencies in CD95. However, in FADD dominant negative (FADDdd) mice, early thymocyte development was retarded and peripheral lymphocyte pools were devoid of normal populations of T cells. We show that thymocytes and peripheral T cells from FADDdd display signaling anomalies, implying that FADD plays a previously uncharacterized role in T cell development and activation