Elicitation of endotoxemic effects in C3H/HeJ mice with glucocorticoid antagonizing factor and partial characterization of the factor

Abstract

C3H/HeJ mice were used to study the origin and nature of endotoxin-induced glucocorticoid antagonizing factor (GAF). In conventional mice GAF is believed to be responsible for a variety of effects that occur as a result of an injection of endotoxin, including the inhibition of hormonal induction of hepatic phosphoenolpyruvate carboxykinase and of glyconeogenesis. Responses in such animals are seen whether the endotoxin is extracted with phenol-water or with trichloroacetic acid. C3H/HeJ mice do not respond (or produce GAF?) after an intravenous injection of phenol-water lipopolysaccharide, but they react normally (produce GAF?) when given a trichloroacetic acid preparation. They also behave the same as conventional animals when injected with serum from poisoned normal mice, especially when the reticuloendothelial system of the donors has been activated by prior injections of Zymosan or heat-killed tubercle bacilli. The C3H/HeJ mice have been used, therefore, as assay animals to establish that peak levels of GAF appear in donor serum about 2 h after an injection of lipopolysaccharide, and it is produced intraperitoneally in C3H/HeJ mice given a mixture of endotoxin and peritoneal exudate cells derived from responder mice. GAF elutes from Sephadex G-200 along with markers of known molecular weight in the region of 100,000 to 200,000. It is inactivated by trypsin and by heating at 75 degrees C for 1 h.

Depressed liver regeneration after partial hepatectomy of germ-free, athymic and lipopolysaccharide-resistant mice

“…Endotoxin-resistant C3H/HeJ mice exhibited a deficiency of IL-1 (371, TNF (38), CSF (39,40) and GAF (41) release together with no febrile response (42) to LPS as compared with endotoxin-sensitive CeH/HeN animals. However, the C3H/HeJ mice did respond appropriately to IL-1 with hypoferremia and neutrophilia (42) and to GAF with antagonism to glucocorticoids (41). These findings indicate that LPS-resistant mice have a defect in cytokine production in response to LPS but not in cytokine activity.…”

Purification and Characteristics of Glucocorticoid Antagonizing Factor in Endotoxemia

“…Berry and his colleagues (13,19,20) demonstrated that endotoxemic mice produce a humoral mediator (GAF) in serum from macrophage and that zymosantreated mice showed higher GAF titers in serum than normal mice after endotoxin challenge. The GAF preparation which they used was partially purified by toxin enhances its release from RES macrophage.…”

“…McCallumn and Berry (18) observed that hepatic gluconeogenesis shows severely impaired glycogen synthesis in endotoxemic animals, and also reported that endotoxin prevents the induction of phosphoenolpyruvate carboxykinase (PEPCK) activity after cortisone administration (5). Berry and his colleagues demonstrated that endotoxin-poisoned mice produced a glucocorticoid antagonizing factor (GAF) which inhibits the gluconeogenesis stimulated by steroid administration and that zymosan-treated mice showed higher GAF titers in serum than normal mice 510 S. SAKAGUCHI AND K. YOKOTA after endotoxin challenge (14,19,20). It was found that this factor is produced within 2 hr of an endotoxin injection and exerts at least some of its effects on hepatocytes during the first few hours.…”

“…It was found that this factor is produced within 2 hr of an endotoxin injection and exerts at least some of its effects on hepatocytes during the first few hours. Moore et al (20) reported that GAF inhibits corticosteroid-induced synthesis of the gluconeogenic enzyme PEPCK in endotoxinpoisoned mice and that GAF was eluted by chromatography on a Sephadex G-200 column along with markers of known molecular weight in the region of 100,000 to 200,000. Tryptophan oxygenase (TO) and tyrosine aminotransferase (TAT) are enzymes which are induced in the liver by treatment with glucocorticoid hormones.…”

Pathogenesis of Malaria and Clinically Similar Conditions

“…To our knowledge, this was first demonstrated (in Joe Berry's laboratory in Austin, Tex.) by showing that C3H/HeJ mice, while refractory to the ability of LPS to block gluconeogenesis, were susceptible to the transfer of this activity via peritoneal exudate cell supernatants from wild-type mice exposed to LPS (285,286). A few years later, the idea of the effects of LPS in mammals being mediated by soluble factors generated by host cells had become a widely accepted principle (273).…”