Hypoxia induces a human macrophage cell line to release tumor necrosis factor-alpha and its soluble receptors in vitro

The Journal of Surgical Research
G ScannellG A Granger

Abstract

Tissue hypoxia following hemorrhage and trauma is a possible initiating factor of the generalized inflammatory response seen after shock. The role of hypoxia in the release from a human monocyte cell line (THP-1) of tumor necrosis factor-alpha (TNF alpha) and its soluble membrane receptors (TNF alpha R) in-vitro is investigated in this study. Flat-bottom plates with 500,000 THP-1 cells/ml were placed in air-tight sealed boxes and exposed to hypoxia (O2 = 1%) or controls (O2 = 9%) for up to 24 hr. Supernatants were tested for TNF alpha, as well as 55- and 75-kDa soluble receptors for TNF alpha, by ELISA. Cell viability was assessed by vital dye uptake and was found to be maintained throughout hypoxic exposure. Medium pH levels were within normal range. In eight experiments conducted in duplicate, minimal change over 24 hr occurred in control samples. Control mean and SD were: TNF alpha = 12.0 +/- 4.2, 55-kDa R = 34.6 +/- 2.03, and 75-kDa R = 38.88 +/- 9.68 pg/ml. During hypoxia, TNF alpha was released as early as the first 30 min of exposure (41.3 +/- 2.3 pg/ml) with a small peak at 1 hr (52 +/- 5.0 pg/ml) and a later more pronounced peak at 18 hr (526 +/- 48 pg/ml). Both 55- and 75-kDa R were released by the hypoxic monocytes; ...Continue Reading

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