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Neisseria meningitidis  (meningococcus) causes invasive diseases such as meningitis or septicaemia.  Ex vivo  infection of human whole blood is a valuable tool to study meningococcal virulence factors and the host innate immune responses. In order to consider effects of cellular mediators, the coagulation cascade must be inhibited to avoid clotting. There is considerable variation in the anticoagulants used among studies of  N .  meningitidis  whole blood infections, featuring citrate, heparin or derivatives of hirudin, a polypeptide from leech saliva. Here, we compare the influence of these three different anticoagulants, and additionally Mg/EGTA, on host innate immune responses as well as on viability of  N .  meningitidis  strains isolated from healthy carriers and disease cases, reflecting different sequence types and capsule phenotypes. We found that the anticoagulants significantly impact on cellular responses and, strain-dependently, also on bacterial survival. Hirudin does not inhibit complement and is therefore superior over the other anticoagulants; indeed hirudin-plasma most closely reflects the characteristics of serum during  N .  meningitidis  infection. We further demonstrate the impact of heparin on complement activation on  N .  meningitidis  and its consequences on meningococcal survival in immune sera, which appears to be independent of the heparin binding antigens Opc and NHBA.

Anticoagulants impact on innate immune responses and bacterial survival in whole blood models of Neisseria meningitidis infection