Tumor necrosis factor measurement

KOning A, Muhlbauer RC, Fleisch H (1988) Tumor necrosis factor alpha and interleukin-1 stimulate bone resorption in vivo as measured by urinary [3H]tetracycline excretion from prelabeled mice. J Bone Miner Res 3 621-627... 

Figure B3.6.9 Quenching of fluorescence of tumor necrosis factor (TNF) on denaturation. Fluorescence emission spectra are shown for native TNF (solid line), guanidine-unfolded TNF (dotted line), and acid-denatured TNF (dashed line). Parameters TNF concentration, 30 pg/ml Xex = 280 nm bandwidths ranging from 16 to 24 nm. Measurements made using Perkin-Elmer MPF3 spectrofluorimeter. Reprinted from Hlodan and Pain (1994) with permission of Elsevier Science.

Komatsu M, Kobayashi D, Saito K, Furuya D, Yagihashi A, Araake H, Tsuji N, Sakamaki S, Niitsu Y, Watanabe N. Tumor necrosis factor-alpha in serum of patients with inflammatory bowel disease as measured by a highly sensitive immuno-PCR. Clin Chem 2001 47(7) 1297-1301. 

Watanabe N. Clinical significance of measurement of circulating tumor necrosis factor alpha. Rinsho Byori 2001 49(9) 829-833. 

Many mediators of inflammation have been identified— cytokines IL-6, tumor necrosis factor alpha cell adhesion molecules intracellular adhesion molecule-1 (ICAM-I), P-selectin and acute phase reactants CR.R fibrinogen, serum amyloid A, and soluble CD40 (Fig. I) (3). Myeloperoxidase is an enzyme secreted from monocytes, neutrophils, and macrophages. A single measurement taken from patient with chest pain in the emergency department predicted the early risk of myocardial infarction and the risk of major cardiac of ends in the next 30 days to six months (15). 

Since oxidative stress has been reported to produce subcellular redistribution and activation of NF-kB, we measured total NF-kB content in cytosolic and particulate fractions of the hearts subjected to ischemia/reperfusion. In another set of experiments, the activation of NF-kB protein was studied by measuring the total NF-kB and phospho-NF-KB in the heart homogenate. Elevated ratios of NF-kB in particulate versus cytosolic fraction and of phospho-NF-KB and total NF-kB in the hearts subjected to ischemia/reperfusion were reduced by selenium. As oxidative stress has been reported to produce subcellular redistribution and activation of NF-kB and antioxidants have been shown to prevent this alteration (Cargnoni et al. 2002), it is possible that our observation on selenium-induced NF-kB translocation might be due to its antioxidant action. It should be noted that since selenium treatment could reduce the tumor necrosis factor-cc (TNF-ct)-mediated activation of NF-kB (Kim and Stadtman 1997), it suggests a possible involvement of a reduction in the formation of TNF-tx in the hearts subjected to ischemia/reperfusion by selenium. 

In this chapter, we have reviewed some critical aspects of stoppers that are used for packaging of lyophilized biopharmaceuticals. We have listed the desirable attributes for lyophilization closures for use with protein formulations. We have discussed issues relating to preparation of stoppers. Two issues, siliconization and occluded water, have been dealt with in detail. Siliconization can be a cause of particulate generation and aggregation, as shown by our experience with recombinant tumor necrosis factor formulations. We have described methods of measurement of stopper-occluded moisture and discussed their relative merits and demerits. We further studied the kinetics of moisture removal from stoppers under selected conditions. Our approach to validation of moisture removal from stoppers has also been presented. Finally, we have discussed the usefulness of occluded moisture measurement for determination of moisture uptake by protein products under real... 

The use of isotope-labelled synthetic peptides as IS was proposed for the absolute quantification of proteins in protein expression studies [113]. If necessary, these synthetic peptides can be covalently modified to be applied as IS in for instance phosphopeptide quantification. This approach was applied in the quantitative analysis of two glutathione 5 -transferase isoforms in human liver cytosol by LC-MS in SRM mode [114]. A series of pilot experiments were performed to select the most suitable IS peptides for four human plasma proteins (hemopexin, ocl antichymotrypsin, interleiddn-6, and tumor necrosis factor-oc) [115]. Rabbit polyclonal antibodies were raised against these selected peptides and immobilized on POROS supports. These lAC columns were applied to achieve a 120-fold emichment of the antigen peptide from digested human plasma proteins. The peptides and its IS were measured by LC-MS in SIM or SRM mode. The methods appears to be a tailor method for targeted protein analysis. 

We are still at the very early stages of interpreting the clinical relevance of endogenous cytokine concentrations. Not only is the immune system affected by cytokines such as IL-1, IL-6, and tumor necrosis factor a (TNF-a), but other systems (skeletal, endocrine, central nervous system) are also affected. Therefore, measurement of... 

Many patients with liver failure have a malabsorption syndrome and chronic diarrhea. Chronic diarrhea causes zinc deficiency because stool contains substantial quantities of zinc. Cytokines such as tumor necrosis factor, interleukin-1, and interleukin-6 may stimulate metal-lothionein, an intestinal zinc-binding protein, thereby inhibiting zinc absorption. Considering the importance of zinc in metalloenzyme reactions, wound healing, immunocompetence, and the senses of taste and smell, patients with chronic diarrhea or large ostomy losses should be suspected of having zinc deficiency measurement of serum concentrations may be used to confirm such deficiencies. Patients receiving a protein-restricted diet may be at additional risk because substantial amounts of zinc are found in red meat. 

Archer, S. (1993) Measurement of nitric oxide in biological models. Faseb J. 7 349-360. Bagavandoss, P., Wiggins, R.C.,Kunkel, S.L.,Remick, D.G., and Keyes, P.L. (1990). Tumor necrosis factor production and accumulation of inflammatory cells in the corpus luteum of pseudopregnancy and pregnancy in rabbits. Biol.Reprod. 42 367-376. 

Fig. 1. Schematic diagram of the major histocompatibility complex on human chromosome 6. Fully expressed genes are shown in black pseudogenes (which do not express a product) are shown in white and genes of undefined or uncertain status are shaded. Distances along the chromosome are measured in units known as kilobase pairs (kbp). BAT indicates B-associated transcript TNF, tumor necrosis factor. Adapted fiom Peter and Hawkins (P3) with permission, Copyright 1992, American Medical Association.

Surcel H, Syrjala H, Karttunen R, Tapaninaho S, Herva E. Development of Francisella tularensis antigen responses measured as T-lymphocyte proliferation and cytokine production (tumor necrosis factor alpha, gamma interferon, and interleukin-2 and -4) during human tularemia. Infect Immun. 1991 59 1948-1953. 

Fig. 2. EPA decreases lipopolysaccharide (LPS)-induced tumor necrosis-factor (TNF)-a production. THP-1 cells preincubated with increased concentrations of eicosapentaenoic acid (EPA) (0, 10, 60, 120 and 180 p,M) for 24 h were stimulated with 0.2 p.g/mL of EPS for 6 h. TNF-a in the cell culture supernatant was measured by enzyme-linked immunosorbent assay. The amount of TNF-a produced in the control EPS-stimulated cells is set as 100%. Values are means + SEM, n = 3.

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