Protocols to Interrogate the Interactions Between Neisseria gonorrhoeae and Primary Human Neutrophils.

Neisseria gonorrhoeae (Gc) infection of its obligate human host results in a robust neutrophil-driven immune response. Despite neutrophils' intrinsic ability to neutralize microbes, Gc can survive in the presence of neutrophils. To interrogate how this pathogen evades killing by neutrophils, we employ an ex vivo model of Gc infection with Interleukin-8-primed and adhered primary human neutrophils. This chapter will describe how primary human neutrophils are purified from venous blood, how Gc is prepared for infection, how to assess Gc survival in the presence of human neutrophils by enumeration of colony forming units, and how to determine Gc internalization by human neutrophils using an immunofluorescence-based approach.