Detection of membrane-bound tumor necrosis factor (TNF): An analysis of TNF-specific reagents

Gerspach, Jeannette; Götz, Alex; Zimmermann, Gudrun; Kolle, Carmen; Böttinger, Heiner; Grell, Matthias

doi:10.1002/1097-0029(20000801)50:3<243::aid-jemt8>3.0.co;2-b

Cited by 21 publications

(20 citation statements)

References 31 publications

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“…Production of TNF has also been proposed as a potential alternate or complementary T cell correlate (45). The findings presented here have important implications for the application of TNF as a correlate; measurement of memTNF is much more difficult technically than measurement of secreted TNF (46). Secreted TNF quantities may not reflect the most relevant T cell activities, but thus far attempts to measure cell surface TNF on activated LVS-immune T cells using flow cytometry have been inconclusive (data not shown).…”

Section: Discussionmentioning

confidence: 86%

Differential Requirements by CD4+ and CD8+ T Cells for Soluble and Membrane TNF in Control of Francisella tularensis Live Vaccine Strain Intramacrophage Growth

Cowley

Sedgwick

Elkins

2007

The Journal of Immunology

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During primary infection with intracellular bacteria, the membrane-associated form of TNF provides some TNF functions, but the relative contributions during memory responses are not well-characterized. In this study, we determined the role of T cell-derived secreted and membrane-bound TNF (memTNF) during adaptive immunity to Francisella tularensis live vaccine strain (LVS). Although transgenic mice expressing only the memTNF were more susceptible to primary LVS infection than wild-type (WT) mice, LVS-immune WT and memTNF mice both survived maximal lethal secondary Francisella challenge. Generation of CD44high memory T cells and clearance of bacteria were similar, although more IFN-γ and IL-12(p40) were produced by memTNF mice. To examine T cell function, we used an in vitro tissue coculture system that measures control of LVS intramacrophage growth by LVS-immune WT and memTNF-T cells. LVS-immune CD4+ and CD8+ T cells isolated from WT and memTNF mice exhibited comparable control of LVS growth in either normal or TNF-α knockout macrophages. Although the magnitude of CD4+ T cell-induced macrophage NO production clearly depended on TNF, control of LVS growth by both CD4+ and CD8+ T cells did not correlate with levels of nitrite. Importantly, intramacrophage LVS growth control by CD8+ T cells, but not CD4+ T cells, was almost entirely dependent on T cell-expressed TNF, and required stimulation through macrophage TNFRs. Collectively, these data demonstrate that T cell-expressed memTNF is necessary and sufficient for memory T cell responses to this intracellular pathogen, and is particularly important for intramacrophage control of bacterial growth by CD8+ T cells.

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Section: Discussionmentioning

confidence: 86%

Differential Requirements by CD4+ and CD8+ T Cells for Soluble and Membrane TNF in Control of Francisella tularensis Live Vaccine Strain Intramacrophage Growth

Cowley

Sedgwick

Elkins

2007

The Journal of Immunology

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“…Polyclonal antimouse TNF (Genzyme Virotech, Rüsselheim, Germany) antibodies (1:100 diluted serum) were visualized with Cy5-conjugated goat antirabbit antibodies (diluted 1:1000; Dianova, Hamburg, Germany). Cryostat sections from the same type of tumor were fixed with cold acetone and stained with TNF receptor-2-human immunoglobin G (TNFR2-huIgG) fusion protein 17,18 (10 g/mL) and visualized using rabbit antihuman IgG antibody (diluted 1:100; Cappel, Durham, NC), biotinylated goat antirabbit IgG antibody (diluted 1:250; Boehringer, Mannheim, Germany), and fluorescein isothiocynate (FITC)-labeled streptavidin (diluted 1:1000; Zymed, San Francisco, CA). Rat antimouse CD31 (1 g/mL; Pharmingen, San Diego, CA) was visualized with tetrarhodamine isothiocyanate-conjugated goat antirat IgG (diluted 1:25; Zymed).…”

Section: Immunohistochemistrymentioning

confidence: 99%

“…Cells were selected for resistance to zeocin, and clones were isolated, transferred into 96-well plates, and analyzed by fluorescence-activated cell sorter (FACS) analysis with the TNF-specific antibody T1. 17 …”

Section: Cell Culture and Transfection Of Endothelial Cellsmentioning

confidence: 99%

A permissive role for tumor necrosis factor in vascular endothelial growth factor-induced vascular permeability

Clauss

Sunderkötter

Sveinbjørnsson

et al. 2001

Blood

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“…Sections were counterstained with Gill's hematoxylin I and mounted with Aquatex (Merck Eurolab, Frankfurt, Germany) or a solution of Mowiol 488 (Calbiochem, Bad Soden, Germany). Staining of liver and lung sections for TNF was performed as previously described (28) using the Ab IP-400 (Genzyme, Ruesselsheim, Germany) or a human sTNFRI/Fc fusion protein (R&D Systems, Wiesbaden, Germany) (29). Isotypematched controls were rabbit normal serum and human IgG, respectively.…”

Section: Immunofluorescence Immunohistochemical Staining and Tunel mentioning

confidence: 99%

Chronic Inflammation and Protection from Acute Hepatitis in Transgenic Mice Expressing TNF in Endothelial Cells

Willuweit

Sass

Schöneberg

et al. 2001

The Journal of Immunology

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Endothelial activation is an important feature of many inflammatory diseases and has been implicated as the cause of vascular complications in disorders such as diabetes, atherosclerosis, and transplant rejection. One of the most potent activators of the endothelium is TNF, which can also be expressed by endothelial cells, causing a permanent, autocrine stimulatory signal. To establish a model of continuous endothelial activation and to elucidate the role of endothelial derived TNF in vivo, we generated transgenic mice expressing a noncleavable transmembrane form of TNF under the control of the endothelial-specific tie2 promoter. Adult tie2-transmembrane TNF-transgenic mice developed chronic inflammatory pathology in kidney and liver, characterized by perivascular infiltration of mononuclear cells into these organs. Along with the infiltrate, an up-regulation of the adhesion molecules ICAM-1 and VCAM-1, but not E-selectin, in the endothelium was observed. Despite predisposition to chronic inflammation these mice were protected from immune-mediated liver injury in a model of Con A-induced acute hepatitis. Although the blood levels of soluble TNF and IFN-γ were increased in transgenic animals after challenge with Con A, no damage of hepatocytes could be detected, as assessed by the lack of increase in plasma transaminase activities and the absence of TUNEL staining in the liver. We conclude that expression of transmembrane TNF in the endothelium causes continuous endothelial activation, leading to both proinflammatory and protective events.

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Detection of membrane-bound tumor necrosis factor (TNF): An analysis of TNF-specific reagents

Cited by 21 publications

References 31 publications

Differential Requirements by CD4+ and CD8+ T Cells for Soluble and Membrane TNF in Control of Francisella tularensis Live Vaccine Strain Intramacrophage Growth

Differential Requirements by CD4+ and CD8+ T Cells for Soluble and Membrane TNF in Control of Francisella tularensis Live Vaccine Strain Intramacrophage Growth

A permissive role for tumor necrosis factor in vascular endothelial growth factor-induced vascular permeability

Chronic Inflammation and Protection from Acute Hepatitis in Transgenic Mice Expressing TNF in Endothelial Cells

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