Neisseria gonorrhoeae (Ngo), the causative agent of gonorrhea, uses lipooligosaccharide (LOS) sialylation as a key mechanism of complement evasion. Since Ngo cannot synthesize its own sialic acid, this process depends on its enzyme sialyltransferase (Lst), which incorporates host-derived cytidine-5’-monophospho-N-acetylneuraminic acid (CMP-NANA), a metabolically activated form of sialic acid, into LOS in its outer membrane. Bacterial vaginosis (BV), frequently associated with Gardnerella vaginalis (Gv), is linked to increased gonococcal infections. Gv produces vaginolysin (VLY), a pore-forming toxin that lyses epithelial cells, releasing intracellular contents, including sialic acid. Ngo utilizes this released sialic acid to sialylate LOS and enhance serum resistance. CD59, a complement inhibitor expressed on epithelial cells, functions as a receptor for VLY and facilitates its association with host cells for its lytic activity. While VLY’s role in promoting Ngo survival is known, the role of CD59 in modulating VLY ability to lyse epithelial cells and its subsequent impact on Ngo serum survival remains poorly understood. This study demonstrates that CD59 enhances VLY-mediated epithelial cytolysis, facilitating the release of intracellular contents including sialic acid which Ngo utilizes for LOS sialylation. Using CRISPR/Cas9-generated CD59 knockout (KO) ME-180 cervical epithelial cells, we showed that CD59 is essential for VLY-induced cytotoxicity, measured by lactate dehydrogenase (LDH) release, trypan blue exclusion, and MTT assays. Serum bactericidal assays demonstrated that supernatants from VLY-treated WT ME-180 cells significantly enhanced Ngo serum survival in an Lst-dependent manner, an effect absent in CD59 KO cells. However, WT Ngo survival relative to a Δlst mutant was restored when treated with supernatants from CD59 rescued ME-180 cells. These findings highlight a role for CD59 in facilitating VLY-mediated epithelial cell lysis, which releases intracellular contents, including sialic acid, enabling Ngo to enhance its serum resistance through LOS sialylation