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Discussion on HSV-2 strains and assembly of mRNA and LNP

Posted by Saher on 04 Jun 2021 at 10:59 GMT

This work is really amazing. I have few points to discuss here:
1. Is there any reason to choose different strains as you have used HSV - 2 strain 333 for Glycoprotein C and Glycoprotein D but for Glycoprotein E you have used HSV - 2 strain 2.12. For mice challenge, again you have used another strain that is HSV - 2 strain MS. Could you please explain why these strains have been selected for vaccination as there are lot of other options as well.
2. You have combined trivalent in LNPs, as these are nanoparticles, how much lengthy mRNA can get fit into it and how do you assure all three mRNA in LNPs.
I would really appreciate if you give any clarification for the better understanding of readers.

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RE: Discussion on HSV-2 strains and assembly of mRNA and LNP

hfriedman replied to Saher on 13 Jul 2021 at 14:54 GMT

1. Regarding HSV-2 strains selected: Approximately 10 years ago we started immunizing mice and guinea pigs with glycoproteins prepared in baculovirus. We were working with the Cohen lab at Penn. His lab had prepared gC2 and gD2 baculovirus proteins from strain 333, while my lab had prepared gE2 from strain 2.12. The gC2, gD2 and gE2 sequences are highly conserved among different HSV-2 isolates. When we started working with nucleoside mRNA we decided to use the identical amino acid sequences as we used for baculovirus so that we could compare the two immunization strategies head-to-head.
2. Regarding length of mRNA: My colleague Drew Weissman has successfully encapsulated up to 10Kb mRNA.
Regarding encapsulating all 3 mRNA: Our goal is to encapsulate all 3, but even if variable, individually encapsulated mRNA is effective.

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