A recombinant tumor necrosis factor-α p80 receptor:Fc fusion protein decreases circulating bioactive tumor necrosis factor-α but not lung injury or mortality during immunosuppression-related gram-negative bacteremia

Purpose: During gram-negative bacteremia (GNB), tumor necrosis factor-α (TNF-α) is a critical early mediator of host defense, whose overexpression can initiate acute lung injury, multiple organ failure, and death. In this study we evaluated the ability of a chimeric fusion protein containing two extracellular domains of the human p80 TNF-α receptor and the Fc region of human IgG 1 (TNFR:Fc) to reduce circulating TNF-α, and to ameliorate organ injury and improve survival in a rodent model of GNB during immunosuppression-related neutropenia. Materials and Methods: Conscious catheterized male rats ( n = 37) with stable cyclophosphamide-induced neutropenia were infected intravenously (IV) with 5 × 10 9 live Escherichia coli (EC, serotype 055:135) ending at t = 0. All animals received antibiotics (penicillin/ amikacin sulfate) at t = 0.5 and t = 8 hours, and 0.9% sodium chloride (normal saline solution (NS), 1 mL/h) from t = 0 to 8 hours. Subgroups were post-treated at t = 0.5 hours with a 1.0 mL IV dose of TNFR:Fc (60, 600, or 1,200 μg; Immunex), 600 μg of human IgG1-κ or IgG1-λ (Sigma), or NS alone (controls). A separate TNFR:Fc pretreatment subgroup received 600 μg/rat of the fusion protein 5 minutes before starting EC infusion. Hemodynamics were monitored continuously through t = 24 hours, and arterial samples were collected at baseline and at t = 1.5,4.5,8, and 24 hours after EC were analyzed for blood gases, quantitative culture, serum endotoxin, bioactive and antigenic TNF-α, and formed elements. Postmortem tissues were examined for histopathologic changes. Results: Compared with antibiotic-treated and fluid-supported controls, TNFR:Fc dose-dependently reduced bioactive but not antigenic TNF-α without altering bacterial clearance, serum endotoxin, or 24-hour survival. Of note, 600 pg of IgGl-κ or IgG1-λ attenuated peak bioactive TNF-α to a similar degree as 1,200 μg TNFR:Fc, and also significantly reduced serum endotoxin levels. Nevertheless, by t = 8 hours all bacteremic rats were hypothermic with tachypnea-related hypocarbia and hyperoxemia and were thrombocytopenic. At death, all subgroups showed similar hepatic glycogen depletion and pulmonary congestion with perivascular edema and alveolar hemorrhage. Conclusions: Although TNFR:Fc and its idiotypic control IgG1 reduced circulating bioactive TNF-α, neither treatment prevented progression of lethal shock with attendant organ injury in this conscious, antibiotic-treated and fluid-resuscitated model of immunosuppression-related GNB.