February 1994

Reduced Tumor Necrosis Factor α Production in Lipopolysaccharide-Treated Whole Blood From Patients in the Intensive Care Unit

Author Affiliations

From the Departments of Surgery (Drs Setrakian, Yee, and Christou) and Microbiology and Immunology (Dr Christou), McGill University, Montreal, Quebec.

Arch Surg. 1994;129(2):187-192. doi:10.1001/archsurg.1994.01420260083011

Objective:  To determine the cytokine response to lipopolysaccharide in patients in the intensive care unit.

Patients:  Patients in a mixed medical/surgical intensive care unit with fever and a de novo clinical dysfunction of at least one organ system.

Methods:  Whole blood from patients and from laboratory controls was stimulated with 8 ng/mL of lipopolysaccharide (Escherichia coli 0111:B4) at 37°C, and tumor necrosis factor α (TNF-α) was measured using enzyme linked immunosorbent assay at 4,8, and 24 hours. The same subjects' purified monocytes were cultured with 8 ng/mL of lipopolysaccharide in the presence of autologous or pooled control plasma or cocultured with purified autologous polymorphonuclear leukocytes at a polymorphonuclear leukocyte—monocyte ratio of 10:1, and TNF-α was measured at 24 hours using the enzyme linked immunosorbent assay.

Results:  We detected high (n=5) and low (n=5) TNF-α responders in whole blood producing a mean (±SEM) of 27.2± 6.3 pg/mL per 1000 monocytes vs 0.0±2.4 pg/mL per 1000 monocytes, respectively (controls, 58.0±13.0 pg/mL per 1000 monocytes). The kinetics of TNF-α production in both groups were comparable. Purified monocytes from both groups of patients cultured with lipopolysaccharide alone produced equivalent TNF-α values (42.4±10.5 vs 40.8± 12.5 pg/mL per 1000 monocytes). Assayable TNF-α was not different with autologous vs control serum but was markedly diminished by the presence of polymorphonuclear leukocytes in patients as well as in controls; the two groups of patients did not differ in this polymorphonuclear leukocyte effect.

Conclusion:  Lipopolysaccharide stimulation of monocytes in the whole blood results in marked variation of TNF-α production. This phenomenon may account for the variable septic response to infection in patients in the intensive care unit.(Arch Surg. 1994;129:187-192)