Effective Immunomodulatory Treatment ofEscherichia coliExperimental Sepsis with Thalidomide

Abstract

Thalidomide, an agent which inhibits biosynthesis of tumor necrosis factor alpha (TNF-␣) and which is used to treat a variety of chronic inflammatory conditions, was investigated as therapy for experimental sepsis. Sepsis was induced by intraperitoneal injection of 10 7 CFU of Escherichia coli per kg of body weight to 80 Wistar rats divided into four groups. Group A consisted of 24 control animals that did not receive any pretreatment, group B consisted of 18 vehicle-treated control animals pretreated with seed oil, group C consisted of 30 rats administered thalidomide diluted in seed oil at a dose of 50 mg/kg 30 min before bacterial challenge, and group D consisted of eight animals that were not challenged with E. coli but that were used for white blood cell count determination. Sepsis was determined by measurement of vital signs before and 6 h after bacterial challenge. After 6 h the animals were killed and blood was sampled for culture; white blood cell count determination; and determination of endotoxin (lipopolysaccharide), TNF-␣, interleukin-1␤ (IL-1␤), and IL-6 levels. The levels of these cytokines were also estimated in the supernatants of human monocytes pretreated with thalidomide after exposure to the isolate. Sepsis developed in all vehicle-treated control animals and controls by 6 h after bacterial challenge but in only 10 animals (33.3%) pretreated with thalidomide (P < 0.0001). Six hours after bacterial challenge all animals had similar levels of endotoxemia, IL-1␤, and IL-6. The mean white blood cell count for groups A, B, and C were 5,631.1, 2,638.9, and 8,169.3 cells/l, respectively (P value between groups, <0.0001); the TNF-␣ levels were 77.3, 107.2, and 26.1 pg/ml, respectively (P values between groups, <0.0001). Pretreatment of human monocytes with thalidomide prevented the secretion of TNF-␣ and IL-1␤ but not that of IL-6. It is concluded that thalidomide exerts a considerable anti-inflammatory effect by preventing evolution to sepsis and by decreasing the level of production of TNF-␣ and therefore deserves to be further evaluated in research for the therapy of sepsis.Over the last decade the idea of inhibiting tumor necrosis factor alpha (TNF-␣) biosynthesis or TNF-␣ activity as an immunomodulatory therapy for sepsis caused by gram-negative organisms has been thoroughly explored with various animals and in clinical trials (4, 18). Results from recent human studies with the p55 tumor necrosis factor receptor protein (lenercept), however, were disappointing (1). Thalidomide is an old agent that in 1998 was approved for use by the Food and Drug Administration for the treatment of erythema nodosum leprosum. Since then, further knowledge on its immunomodulatory properties has been acquired. A major effect of this drug is reduction of the half-life of TNF-␣ mRNA in human monocytes (7). Thalidomide is used to treat a variety of dermatological conditions, such as cutaneous lupus erythematosus and Behçet's syndrome; various conditions associated with human immunodeficiency virus infection like apht...

Immunomodulatory Clarithromycin Treatment of Experimental Sepsis and Acute Pyelonephritis Caused by Multidrug-Resistant Pseudomonas aeruginosa

“…Pretreatment with potent immunomodulatory agents in animal studies may have led to the failure of previous clinical trials (1). Administration of clarithromycin in parallel with bacterial challenge allows (i) avoidance of pretreatment; (ii) the ability to achieve the desired drug levels within 30 to 60 min after intravenous infusion, i.e., later than bacterial inoculation; and (iii) documentation of whether the secretion of proinflammatory mediators is prevented, which constitutes the necessary step before proceeding to drug administration in the event of the presence of clinical symptoms of sepsis (9).…”

“…A total of 10 8 CFU of the P. aeruginosa isolate per kg in a volume of 0.1 ml was injected into the renal pelvis, proximal to the suture, with a 26-gauge needle. The bacterial inoculum was selected on the basis of those used in previous studies (9). The peritoneal cavity and the abdominal wall were then closed in layers.…”

Angiopoietin-2 Enhances Survival in Experimental Sepsis Induced by Multidrug-ResistantPseudomonas aeruginosa

“…As such, inhibition of TNF-␣ has provided prolonged survival in a variety of animal models of sepsis induced after challenge with LPS or Gram-negative bacteria such as E. coli (Tracey et al, 1987;Giamarellos-Bourboulis et al, 2003). The biological significance of TNF-␣ as the acute proinflammatory cytokine leading to early death has been shown in experiments using TNF-deficient mice.…”

Effect of Thalidomide on Endotoxin-Induced Decreases in Activity and Expression of Hepatic Cytochrome P450 3A2

“…[17][18][19][20][21] There are interesting reports suggesting that thalidomide improves endotoxin-or alcohol-induced liver injury, 22,23) and prolongs the survival rate following the experimental sepsis induced by endotoxin or bacterial challenge in rats. 24,25) Shimazawa et al 26) have reported that thalidomide and its analogs have NO synthase (NOS)-inhibitory activity in vitro, but the activity of thalidomide is weak. Enomoto et al 27) reported that thalidomide abolishes the endotoxin-induced increase in CD14 expression in Kupffer cells in vitro.…”

“…The dose of thalidomide used in this study was chosen on the basis of that used in previous studies. 25) Twenty-four hours after injection of endotoxin or saline, antipyrine clearance experiments were conducted, since we previously reported that the level of endotoxin-induced decrease in hepatic drug-metabolizing enzyme activity reaches the maximum 24 h after endotoxin administration. 29) Rats received a bolus intravenous injection of antipyrine (20 mg/kg of body weight), which was dissolved in saline at the concentration of 10 mg/ml.…”