A novel sialylation site on Neisseria gonorrhoeae lipooligosaccharide links heptose II lactose expression with pathogenicity

Ram, S.; Gulati, S.; Lewis, L. A.; Chakraborti, S.; Zheng, B.; DeOliveira, R. B.; Reed, G. W.; Cox, A. D.; Li, J.; St. Michael, F.; Stupak, J.; Su, X.-H.; Saha, S.; Landig, C. S.; Varki, A.; RICE, P. A.

doi:10.1101/302968

bioRxiv · 10.1101/302968

A novel sialylation site on Neisseria gonorrhoeae lipooligosaccharide links heptose II lactose expression with pathogenicity

Ram, S.·Gulati, S.·Lewis, L. A.·Chakraborti, S.·Zheng, B.·DeOliveira, R. B.·Reed, G. W.·Cox, A. D.·Li, J.·St. Michael, F.·Stupak, J.·Su, X.-H.·Saha, S.·Landig, C. S.·Varki, A.·RICE, P. A.

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Abstract

Sialylation of lacto-W-neotetraose (LNnT) extending from heptose I (HepI) of gonococcal lipooligosaccharide (LOS) contributes to pathogenesis. Previously, gonococcal LOS sialyltransterase (Lst) was shown to sialylate LOS in Triton X-100 extracts of strain 15253, which expresses lactose from both HepI and HepII, the minimal structure required for mAb 2C7 binding. Ongoing work has shown that growth of 15253 in cytidine monophospho-W-acetylneuraminic acid (CMP-Neu5Ac)-containing media enables binding to CD33/Siglec-3, a cell surface receptor that binds sialic acid, suggesting that lactose termini on LOS of intact gonococci can be sialylated. Neu5Ac was detected on LOSs of strains 15253 and a MS11 mutant with only lactose from HepI and HepII by mass spectrometry; deleting HepII lactose rendered Neu5Ac undetectable. Resistance of HepII lactose Neu5Ac to desialylation by 2-3-specific neuraminidase suggested an 2-6-linkage. Although not associated with increased factor H binding, HepII lactose sialylation inhibited complement C3 deposition on gonococci. 15253 mutants that lacked Lst or HepII lactose were significantly attenuated in mice, confirming the importance of HepII Neu5Ac in virulence. All 75 minimally passaged clinical isolates from Nanjing, China, expressed HepII lactose, evidenced by reactivity with mAb 2C7; mAb 2C7 was bactericidal against the first 62 (of 75) isolates that had been collected sequentially and were sialylated before testing. mAb 2C7 effectively attenuated 15253 vaginal colonization in mice. In conclusion, this novel sialylation site could explain the ubiquity of gonococcal HepII lactose in vivo. Our findings reiterate the candidacy of the 2C7 epitope as a vaccine antigen and mAb 2C7 as an immunotherapeutic antibody.

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Ram, S., Gulati, S., Lewis, L. A., Chakraborti, S., Zheng, B., DeOliveira, R. B., Reed, G. W., Cox, A. D., Li, J., St. Michael, F., Stupak, J., Su, X.-H., Saha, S., Landig, C. S., Varki, A., RICE, P. A.. 2018-04-17. A novel sialylation site on Neisseria gonorrhoeae lipooligosaccharide links heptose II lactose expression with pathogenicity. https://doi.org/10.1101/302968

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