𝐃𝐞𝐚𝐫 𝐏𝐫𝐢𝐲𝐚𝐥,𝐲𝐨𝐮 𝐡𝐚𝐯𝐞 𝐚 𝐠𝐨𝐨𝐝 𝐪𝐮𝐞𝐬𝐭𝐢𝐨𝐧. Neisseria gonorrhoeae causes the sexually transmitted disease gonorrhoea by evading innate immunity. Colonizing the mucosa of the reproductive tract depends on the bacterial outer membrane porin, PorB, which is essential for ion and nutrient uptake. PorB is also targeted to host mitochondria and regulates apoptosis pathways to promote infections. How PorB traffics from the outer membrane of N. gonorrhoeae to mitochondria and whether it modulates innate immune cells, such as macrophages, remains unclear. Here, we show that N. gonorrhoeae secretes PorB via outer membrane vesicles (OMVs). Purified OMVs contained primarily outer membrane proteins including oligomeric PorB. The porin was targeted to mitochondria of macrophages after exposure to purified OMVs and wild type N. gonorrhoeae. This was associated with loss of mitochondrial membrane potential, release of cytochrome c, activation of apoptotic caspases and cell death in a time-dependent manner. Consistent with this, OMV-induced macrophage death was prevented with the pan-caspase inhibitor, Q-VD-PH. This shows that N. gonorrhoeae utilizes OMVs to target PorB to mitochondria and to induce apoptosis in macrophages, thus affecting innate immunity.

𝐅𝐨𝐫 𝐦𝐨𝐫𝐞 𝐢𝐧𝐬𝐢𝐠𝐡𝐭𝐬 𝐫𝐞𝐠𝐚𝐫𝐝𝐢𝐧𝐠 𝐲𝐨𝐮𝐫 𝐪𝐮𝐞𝐫𝐲 𝐲𝐨𝐮 𝐜𝐚𝐧 𝐯𝐢𝐯𝐢𝐝𝐥𝐲 𝐠𝐨 𝐭𝐡𝐫𝐨𝐮𝐠𝐡 𝐭𝐡𝐞 𝐟𝐨𝐥𝐥𝐨𝐰𝐢𝐧𝐠 𝐚𝐫𝐭𝐢𝐜𝐥𝐞 𝐨𝐫 𝐥𝐢𝐧𝐤𝐬 𝐛𝐞𝐥𝐨𝐰

𝐀𝐫𝐭𝐢𝐜𝐥𝐞-1: Outer membrane vesicles from Neisseria gonorrhoeae target PorB to mitochondria and induce 𝐚𝐩𝐨𝐩𝐭𝐨𝐬𝐢𝐬

𝐃𝐎𝐈:PLoS Pathog. 2018 Mar; 14(3): e1006945.

Published online 2018 Mar 30. doi: 10.1371/journal.ppat.1006945

PMCID: PMC5877877

PMID: 29601598

𝐀𝐫𝐭𝐢𝐜𝐥𝐞 2:

2019

Springer Protocols

Isolation of Naturally Released Gonococcal Outer Membrane Vesicles as Vaccine 𝐀𝐧𝐭𝐢𝐠𝐞𝐧𝐬

𝐃𝐨𝐢: DOI

https://doi.org/10.1007/978-1-4939-9496-0_9

𝐑𝐞𝐠𝐚𝐫𝐝𝐬,

𝐌𝐃. 𝐎𝐬𝐢𝐦 𝐀𝐪𝐮𝐚𝐭𝐚𝐫 ,𝐂𝐒𝐈𝐑-𝐍𝐄𝐄𝐓𝐈,𝐈𝐍𝐃𝐈𝐀

𝐃𝐞𝐚𝐫 𝐏𝐫𝐢𝐲𝐚𝐥,𝐲𝐨𝐮 𝐡𝐚𝐯𝐞 𝐚 𝐠𝐨𝐨𝐝 𝐪𝐮𝐞𝐬𝐭𝐢𝐨𝐧. Neisseria gonorrhoeae causes the sexually transmitted disease gonorrhoea by evading innate immunity. Colonizing the mucosa of the reproductive tract depends on the bacterial outer membrane porin, PorB, which is essential for ion and nutrient uptake. PorB is also targeted to host mitochondria and regulates apoptosis pathways to promote infections. How PorB traffics from the outer membrane of N. gonorrhoeae to mitochondria and whether it modulates innate immune cells, such as macrophages, remains unclear. Here, we show that N. gonorrhoeae secretes PorB via outer membrane vesicles (OMVs). Purified OMVs contained primarily outer membrane proteins including oligomeric PorB. The porin was targeted to mitochondria of macrophages after exposure to purified OMVs and wild type N. gonorrhoeae. This was associated with loss of mitochondrial membrane potential, release of cytochrome c, activation of apoptotic caspases and cell death in a time-dependent manner. Consistent with this, OMV-induced macrophage death was prevented with the pan-caspase inhibitor, Q-VD-PH. This shows that N. gonorrhoeae utilizes OMVs to target PorB to mitochondria and to induce apoptosis in macrophages, thus affecting innate immunity.

𝐅𝐨𝐫 𝐦𝐨𝐫𝐞 𝐢𝐧𝐬𝐢𝐠𝐡𝐭𝐬 𝐫𝐞𝐠𝐚𝐫𝐝𝐢𝐧𝐠 𝐲𝐨𝐮𝐫 𝐪𝐮𝐞𝐫𝐲 𝐲𝐨𝐮 𝐜𝐚𝐧 𝐯𝐢𝐯𝐢𝐝𝐥𝐲 𝐠𝐨 𝐭𝐡𝐫𝐨𝐮𝐠𝐡 𝐭𝐡𝐞 𝐟𝐨𝐥𝐥𝐨𝐰𝐢𝐧𝐠 𝐚𝐫𝐭𝐢𝐜𝐥𝐞 𝐨𝐫 𝐥𝐢𝐧𝐤𝐬 𝐛𝐞𝐥𝐨𝐰

𝐀𝐫𝐭𝐢𝐜𝐥𝐞-1: Outer membrane vesicles from Neisseria gonorrhoeae target PorB to mitochondria and induce 𝐚𝐩𝐨𝐩𝐭𝐨𝐬𝐢𝐬

𝐃𝐎𝐈:PLoS Pathog. 2018 Mar; 14(3): e1006945.

Published online 2018 Mar 30. doi: 10.1371/journal.ppat.1006945

PMCID: PMC5877877

PMID: 29601598

𝐀𝐫𝐭𝐢𝐜𝐥𝐞 2:

2019

Springer Protocols

Isolation of Naturally Released Gonococcal Outer Membrane Vesicles as Vaccine 𝐀𝐧𝐭𝐢𝐠𝐞𝐧𝐬

𝐃𝐨𝐢: DOI

https://doi.org/10.1007/978-1-4939-9496-0_9

𝐑𝐞𝐠𝐚𝐫𝐝𝐬,

𝐌𝐃. 𝐎𝐬𝐢𝐦 𝐀𝐪𝐮𝐚𝐭𝐚𝐫 ,𝐂𝐒𝐈𝐑-𝐍𝐄𝐄𝐓𝐈,𝐈𝐍𝐃𝐈𝐀

Thank you for your answer Md Osim Aquatar

I appreciate your help. We plan to explore the potential of outer membrane vesicles to be used as vaccine antigens. Thanks much!