The effects of anti-inflammatory and antiallergic drugs on the release of IL-1β and TNF-α in the human whole blood assay

Abstract

Heparinized human whole blood was evaluated as a model to study the effects of various classes of anti-inflammatory drugs on IL-1 beta and TNF-alpha release from leukocytes. Human whole blood was stimulated with zymosan (1.5 mg/ml) or LPS (5 micrograms/ml) to induce significant cytokine release. As previously reported, the 5-lipoxygenase/cyclooxygenase (5-LO/CO) inhibitor, SKF86002 (30 microM), significantly inhibited both IL-1 beta and TNF-alpha release using either stimulus. In contrast, the cyclooxygenase (CO) inhibitors (naproxen and ibuprofen) and the lipoxygenase (5-LO) inhibitors (zileuton, L-663536 and BWA4C) did not effect IL-1 beta or TNF-alpha release/biosynthesis. Isoproterenol (beta-agonist), rolipram (a PDE-IV inhibitor), and IBMX (a nonselective PDE inhibitor), significantly inhibited TNF-alpha but not IL-1 beta in the LPS model while having no effect in the zymosan model. This human whole blood assay is a unique and rapid method which can be used to identify novel inhibitors of IL-1 beta and TNF-alpha release/biosynthesis.

Inhibitory effects of JTE-522, a novel prostaglandin H synthase-2 inhibitor, on adjuvant-induced arthritis and bone changes in rats

“…In our preliminary test, JTE-522 suppressed lipopolysaccharide (LPS)-induced tumor necrosis factor-a (TNF-a) production from human peripheral blood mononuclear cells, and inhibited carbonic anhydrase activity in in vitro experiments (IC 50 value was nanomole order). On the other hand, indomethacin enhanced LPS-induced TNF-a production, which is in agreement with the literature [22], and also activates carbonic anhydrase [23]. TNF-a may be one of the cytokines involved in the bone destruction in adjuvantinduced arthritic joints [24,25] and rheumatic joints [26] while carbonic anhydrase is a key enzyme affecting bone resorption [27,28].…”

Adaptive responses of human monocytes infected byBordetella pertussis: The role of adenylate cyclase hemolysin

“…Downregulation of TNF-␣ by AC-Hly represents a peculiar virulence strategy, in sharp contrast with known effects of most bacterial exotoxins, and has not yet been demonstrated. Inhibition of TNF-␣ release has been demonstrated with different agents that elevate cAMP, including prostaglandin E 2 (Kunkel, 1988), dibutyryl cAMP, cholera toxin (Tannenbaum and Hamilton, 1989), or phosphodiesterase inhibitors (Hartman et al, 1993). Because these agents are known to modulate intracellular cAMP concentrations by distinct mechanisms, it is likely that inhibition of TNF-␣ release results only from changes in cAMP levels.…”

The effects of phosphodiesterase type 4 inhibitors on tumour necrosis factor‐α and leukotriene B4 in a novel human whole blood assay

“…Most of the published methods for TNF-a induction and inhibition by various agents in human whole blood use a 4 h LPS incubation period (Hartman et al, 1993;. To determine whether 4 h is optimal for characterizing PDE4 inhibitors, we examined the e ects of LPS incubation time and PGE 2 on TNF-a inhibition.…”

“…Inhibition of this enzyme leads to increases in intracellular cyclic AMP which can subsequently activate protein kinase A (PKA) (Torphy & Undem, 1991;Seldon et al, 1995). It has been shown with various PDE4-speci®c inhibitors that an increase in intracellular cyclic AMP can inhibit the expression and release of several in¯ammatory mediators, prevent superoxide anion formation in neutrophils, inhibit eosinophil chemotaxis and degranulation as well as lymphocyte proliferation, and causes relaxation of bronchial smooth muscles resulting in bronchodilatation (Nielson et al, 1990;Hartman et al, 1993;Underwood et al, 1994;Schudt et al, 1995;Essayan et al, 1997). PDE4 is predominantly expressed in monocytes, neutrophils and eosinophils (Schudt et al, 1995) and its inhibition could be bene®cial in treating the underlying in¯ammation associated with asthma.…”