Anaerobic growth and cytidine 5'-monophospho-N-acetylneuraminic acid act synergistically to induce high-level serum resistance in Neisseria gonorrhoeae

Abstract

In vivo, gonococci encounter a myriad of conditions not present in vitro. At some stages of infection and disease, gonococci may grow anaerobically, probably by using sodium nitrite as a terminal electron acceptor. Also, gonococci sialylate their lipooligosaccharide (LOS) in vivo, by using low concentrations of cytidine 5'-monophospho-N-acetylneuraminic acid (CMP-NANA) present in host tissue. This sialylation is responsible for the acquired resistance of gonococci to both normal and immune human serum. Given that gonococci grown in the absence of oxygen or in the presence of CMP-NANA probably more closely resemble gonococci grown inside a human host, we studied the serum resistance of gonococci cultivated under these conditions. In the absence of CMP-NANA, anaerobically grown (anaerobic) gonococci were somewhat less sensitive to serum killing than were aerobically grown (aerobic) gonococci. However, anaerobic gonococci grown with 6 micrograms of CMP-NANA per ml exhibited almost complete serum resistance, while aerobic gonococci required 16-fold-higher CMP-NANA concentrations to achieve significant serum resistance. Anaerobic gonococci incubated in CMP-NANA converted to serum resistance two to three times faster than did similarly treated aerobic gonococci and incorporated up to six times as much sialic acid into their LOS. Gonococci can express several different LOS molecules. Anaerobic gonococci expressed the LOS molecule that acts as an acceptor for sialic acid from CMP-NANA in greater quantity than aerobic gonococci did. Finally, Triton X-100 extracts of anaerobic gonococci contained about four times more sialyltransferase activity than did extracts of aerobic gonococci. Sialyltransferase activity in these extracts was not inhibited by oxygen or enhanced by anaerobiosis. These data indicate that anaerobic conditions lead to altered LOS biosynthesis and to induction of sialyltransferase activity in gonococci. In vivo, where decreased oxygen levels and relevant concentrations of CMP-NANA are found, gonococci could readily become resistant to killing by normal and immune human serum.