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17 March 2003

A human whole blood model of LPS-mediated suppression of T cell activation.

Sheraz Yaqub, Vigdis Solhaug, Torkel Vang, Rigmor Solberg, Ansgar O. Aasen, Kjetil Taskén, Jacob E. Wang

Med Sci Monit 2003; 9(3): BR120-126 :: ID: 4736


BACKGROUND: Lipopolysaccharide (LPS) is the main initiator of the early signaling events leading to sepsis caused by Gram-negative bacteria. Late stages of sepsis are associated with impairments of T lymphocyte function, a condition associated with nosocomial infection and poor outcome. The molecular basis for septic immunosuppression is not fully understood. MATERIAL/METHODS: Human whole blood was incubated ex vivo with purified LPS. Cytokine responses and T cell proliferation were assessed, and the role of cyclic 3',5'-adenosine monophosphate (cAMP) in T cell suppression by LPS was studied using a cAMP-antagonist RESULTS: Adding LPS (0.01 to 10 mg/ml) to human blood ex vivo caused a release of prostaglandin E2 (PGE2) in a concentration- and time-dependent manner, with maximal levels of PGE2 obtained with 10 mg LPS per ml blood after 10 hours of incubation. Adding PGE2-concentrations ranging from 0.03 to 10 mM to purified T cells completely abrogated T cell activation and proliferative response, which was largely reversed by adding Rp-8-Br-cAMPS. Peripheral blood mononuclear cells (PBMC) and T cells harvested from whole blood cultured in the presence of LPS ex vivo showed attenuated proliferative response (30-70%) (purified T cells and PBMC) and reduced IL-2 production (85%) upon T cell receptor (TCR)/CD3 activation with anti-CD3. The proliferation in T cells and PBMC was in part restored by Rp-8-Br-cAMPS. CONCLUSIONS: The human whole blood model of LPS-mediated T lymphocyte suppression described in this paper is time and cost efficient, as well as easy to use.

Keywords: 8-Bromo Cyclic Adenosine Monophosphate - analogs & derivatives, Cyclic AMP - blood, Cyclic AMP-Dependent Protein Kinases - blood, Dinoprostone - blood

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750