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Characterization of the coupled plasma filtration–adsorption resin cartridge adsorptive capacity for cytokines and inflammatory mediators: adsorption profiles from septic patient plasma and in vitro endotoxin-stimulated whole blood

Introduction

Coupled plasma filtration–adsorption (CPFA) is an extracorporeal therapy that uses plasma filtration associated with an adsorbent cartridge and hemofiltration in postdilution to remove cytokines and inflammatory mediators associated with septic shock and severe sepsis. We evaluated the adsorptive capacity of the resin cartridge to remove various inflammatory mediators and cytokines in vitro and ex vivo.

Methods

In vitro experiments included static and dynamic evaluations of resin binding. Whole human blood was stimulated with endotoxin for 4 hours at 37°C; or with the addition of added cytokines or toxins for evaluation. For static conditions, 4 ml plasma with 1 ml resin were incubated for up to 10 hours. Aliquots were withdrawn between 0 and 10 hours and cytokine inflammatory mediator and toxin adsorption were determined with standard ELISAs, multianalyte protein arrays, HPLC and diode array adsorption spectroscopy. Dynamic conditions involved defining the optimal linear velocity and evaluating the adsorption capacity under flow conditions. These experiments used a closed circuit consisting of a plasma filter and resin cartridge. Samples were taken from a blood port and immediately before and after the plasma cartridge. In addition, serum, pre-cartridge and post-cartridge plasma samples were also taken from septic patients undergoing CPFA.

Results

Endotoxin-stimulated blood or samples from septic patients had high levels of cytokines and inflammatory mediators. The resin used in the CPFA adsorptive cartridge showed higher than 80% adsorption under both static and dynamic conditions for: IL-1α, IL-6, IL-8, MIP-1α and MIP-1β, TNFα, MCP, myoglobin. IL-6 appeared to be particularly adsorbed by the cartridge. Severe septic patients had great variability and often very high levels of IL-6 ranging from normal levels (50 pg/ml) up to 12,300 pg/ml. The mean of 10 patients treated before CPFA was 1,775 ± 3757 pg/ml, while post-session IL-6 was 995 ± 2178 pg/ml. The plasma levels before the cartridge ranged from 12 pg/ml to 1,750 pg/ml, while post-cartridge levels were below the level of detection.

Conclusion

The resin in CPFA has a high adsorption capacity for several cytokines and mediators involved in severe sepsis and septic shock. Studies are currently ongoing to correlate cytokine reduction with clinically relevant improvements in these patients.

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Livigni, S., Silengo, D., Maio, M. et al. Characterization of the coupled plasma filtration–adsorption resin cartridge adsorptive capacity for cytokines and inflammatory mediators: adsorption profiles from septic patient plasma and in vitro endotoxin-stimulated whole blood. Crit Care 11, P118 (2007). https://0-doi-org.brum.beds.ac.uk/10.1186/cc5278

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Keywords

  • Adsorption Capacity
  • Septic Shock
  • Severe Sepsis
  • Inflammatory Mediator
  • Septic Patient