Original article: cardiovascular
Artificial surface-induced cytokine synthesis: effect of heparin coating and complement inhibition

https://doi.org/10.1016/j.athoracsur.2004.02.005Get rights and content

Abstract

Background

Contact between blood and artificial surfaces induces an inflammatory response including activation of leukocytes and platelets, as well as complement and other plasma cascade systems. In the present study we investigated the roles of complement and surface modification in polyvinylchloride-induced cytokine production.

Methods

Human whole blood was incubated in rotating loops of polyvinylchloride or heparin-coated polyvinylchloride tubing for 4 hours. Plasma concentrations of the cytokines tumor necrosis factor α, interleukin (IL) 1β, IL-6, IL-8, IL-10, and monocyte chemoattractant protein 1 (MCP-1) were quantified.

Results

Polyvinylchloride induced a substantial increase in IL-8 and MCP-1, which was abolished by cycloheximide, indicating that they were synthesized during incubation. Interleukin 8 synthesis was completely complement-dependent since it was abolished by neutralizing antibodies to factor D and complement factor 5, as well as by a complement factor 5a receptor antagonist. Monocyte chemoattractant protein 1 synthesis was reduced by approximately half the amount by the complement inhibitors. Heparin-coated polyvinylchloride efficiently prevented synthesis of both IL-8 and MCP-1. Addition of recombinant human complement factor 5a to blood incubated in heparin-coated polyvinylchloride restored IL-8 and MCP-1 production completely and partly, respectively. In contrast to IL-8 and MCP-1, tumor necrosis factor α, IL-1β, interleukin 6 and IL-10 increased only marginally. A minor but significant increase in IL-1β was complement-dependent, whereas a similar increase in IL-10 was completely prevented by heparin-coated polyvinylchloride. No significant changes were observed for tumor necrosis factor α and IL-6.

Conclusions

Polyvinylchloride induced a marked increase in IL-8 and MCP-1, in contrast to a marginal increase in tumor necrosis factor α, IL-1β, IL-6, and IL-10. The increase in IL-8 and MCP-1 was prevented by heparin-coated polyvinylchloride. Interleukin 8 production was totally complement-dependent and mediated by complement factor 5a.

Section snippets

Reagents

Heparin-coated polyvinyl chloride (H-PVC) tubing (CBAS [Carmeda BioActive Surface] and uncoated PVC tubing were provided by Carmeda AB (Stockholm, Sweden). Sterile phosphate-buffered saline was from Life Technologies (Paisley, UK) and lepirudin (Refludan) from Hoechst (Frankfurt am Main, Germany). Cycloheximide and recombinant human C5a were purchased from Sigma-Aldrich (St. Louis, MO).

Interleukin-8

Incubation of blood for 4 hours in PVC loops increased median IL-8 concentration from 5 to 513 pg/mL (Fig 1, left panel, p < 0.001). This increase was reduced to 126 pg/mL by addition of cycloheximide (Fig 1, left panel, p < 0.02 versus PVC), indicating that the majority of IL-8 was synthesized during incubation.

Monocyte chemoattractant protein 1

Incubation of blood for 4 hours in PVC loops increased median MCP-1 concentration from 47 to 233 pg/mL (Fig 1, right panel, p < 0.001). This increase was reduced to 76 pg/mL by

Comment

The systemic inflammatory response during and after CPB has been a subject of considerable research. That is due both to the frequency with which CPB is performed and its potentially serious complications, as well as to the fact that CPB represents a fairly standardized model for studying in vivo human inflammatory reactions. However, as several factors influence the type and degree of the inflammatory response seen in such settings, results from different studies may not be directly

Acknowledgements

Excellent technical assistance was provided by Hilde Fure and Dorte Christiansen. Financial support was kindly provided by Tanox, Inc, Carmeda AB, The Norwegian Council on Cardiovascular Disease, The Norwegian Foundation for Health and Rehabilitation, and the following legacies: The Blix' family, Odd Fellow, Mariane og Rolf Bjørn, Sparebanken Nord-Norge, and The Sonneborn Chairtable Trust.

References (30)

  • J.B Steinberg et al.

    Cytokine and complement levels in patients undergoing cardiopulmonary bypass

    J Thorac Cardiovasc Surg

    (1993)
  • S Mocellin et al.

    The dual role of IL-10

    Trends Immunol

    (2003)
  • S Wan et al.

    Heparin-coated circuits reduce myocardial injury in heart or heart-lung transplantationa prospective, randomized study

    Ann Thorac Surg

    (1999)
  • S Salek-Ardakani et al.

    Heparin and heparan sulfate bind interleukin-10 and modulate its activity

    Blood

    (2000)
  • Levy JH, Tanaka KA. Inflammatory response to cardiopulmonary bypass. Ann Thorac Surg...
  • Cited by (39)

    • Avoiding ambient air in test tubes during incubations of human whole-blood minimizes complement background activation

      2020, Journal of Immunological Methods
      Citation Excerpt :

      This was made possible by anticoagulating the blood with the thrombin-specific inhibitor lepirudin, which does not interfere with the complement system when collecting blood for in vitro experiments. Undesired background activation of complement and other systems of inflammation in experimental whole-blood models is a common problem during in vitro studies (Lappegård et al. 2004), potentially masking biologically relevant signals in the experiments. In 1980, Track and colleagues discovered that the C3 molecule contained an internal thioester bond.

    • Complement links platelets to innate immunity

      2018, Seminars in Immunology
      Citation Excerpt :

      Furthermore, lethal arterial thrombosis induced by extracellular histones seems to be reduced in C5-deficient mice [129]. Platelet-leukocyte aggregate formation is highly relevant for extracorporeal circulation, which is mediated by C5a [130,131]. Platelet-leukocyte aggregate formation, which is considered a major event in various settings of thrombo-inflammation, is mediated by the alternative complement pathway and specifically its regulator properdin [77,132].

    • The induction of cytokines by polycation containing microspheres by a complement dependent mechanism

      2013, Biomaterials
      Citation Excerpt :

      Among these, the strongest complement dependencies are found for IL-8, MIP-1α and VEGF. The most consistent and strong response is found for the chemokine IL-8 [16,17,19]. We would therefore point to IL-8 as an important mediator for predicting the biomaterial tolerability.

    • Alginate microbeads are complement compatible, in contrast to polycation containing microcapsules, as revealed in a human whole blood model

      2011, Acta Biomaterialia
      Citation Excerpt :

      In such studies bioincompatibility has been measured as overgrowth reactions that might have been caused by inflammatory reactions. The complement system is a primary inductor of inflammation were its protein effectors reacts upstream of leukocytes and cytokines [4,23,35–37]. Complement activity may therefore be a useful parameter for revealing bioincompatibility.

    View all citing articles on Scopus
    View full text