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angie_rasmussen's profile
Dr. Angela Rasmussen
Dr. Angela Rasmussen
Dr. Angela Rasmussen
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@angie_rasmussen

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Dr. Angela RasmussenVerified account

@angie_rasmussen

Excessively direct virologist. PI @VIDOInterVac. Affiliate @georgetown_ghss. Virus-host dramatics. 1X Jeopardy! loser. 🇺🇸in 🇨🇦. Rep: @anniescranton. she/her

Saskatoon, Saskatchewan
angelarasmussen.org
Joined October 2011

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    Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

    Они обалдели! The Sputnik V vaccine Ad5 vector is evidently replication competent. The makers apparently neglected to delete E1, so getting this vaccine means being infected with live adenovirus 5. Hence Brazil’s regulator correctly rejected it.https://www.ctvnews.ca/health/coronavirus/brazil-health-regulator-rejects-russia-s-sputnik-vaccine-1.5403539 …

    6:24 AM - 28 Apr 2021
    • 3,756 Retweets
    • 7,297 Likes
    • Thota Naveen Yuvasena Till Ulenspigel AnaG SpaceFan 🇺🇸 Aditya Leonard T. Muranaga John Makar Charmaine M. Avena Mark
    354 replies 3,756 retweets 7,297 likes
      1. New conversation
      2. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        Adenovirus-vectored vaccines like J&J, AstraZeneca, and Sputnik V use infection with adenoviruses, common pathogens that usually cause common colds. But to increase safety and decrease risk of side effects, most vaccine vectors delete the AdV E1 and E3 genes.

        16 replies 237 retweets 1,239 likes
        Show this thread
      3. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        E1/E3 deletions are standard in Ad-vectors. Deleting E1 prevents the virus from replicating and deleting E3 prevents it from interacting with the immune system.

        4 replies 161 retweets 962 likes
        Show this thread
      4. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        This results in a vaccine vector that causes a transient infection, due to the vector’s inability to complete its replication cycle. When I got J&J it infected my cells, began expressing viral proteins, including SARS-CoV-2 spike, and then hit a block because of the E1 deletion.

        8 replies 136 retweets 896 likes
        Show this thread
      5. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        Sputnik V is a heterologous Ad-vectored vaccine: the prime dose is an Ad26-vectored vaccine (like J&J), followed by a booster that uses Ad5. This was apparently found in testing the Ad5 booster but I can’t emphasize how sloppy this is.

        11 replies 177 retweets 936 likes
        Show this thread
      6. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        When making adenovirus-vectored vaccines, once you’ve deleted a region of the genome, it doesn’t just reappear out of nowhere. The presence of replicating virus suggests E1 was either not deleted or it recombined during manufacturing with a full length AdV genome.

        9 replies 161 retweets 892 likes
        Show this thread
      7. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        And while strong statements from Gamaleya and the RDIF dispute the findings, this isn’t the first observation that the Sputnik V doses submitted to regulators aren’t consistent with the phase 3 trial results published in Lancet.

        6 replies 202 retweets 994 likes
        Show this thread
      8. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        Slovakia’s drug regulator previously rejected a batch of Sputnik V for being substantively different than the batches reported in Lancet. The EMA has also said they have insufficient safety and QC data to authorize it in the EU.https://www.washingtonpost.com/world/2021/04/27/sputnik-vaccine-brazil-russia-coronavirus/ …

        16 replies 346 retweets 1,146 likes
        Show this thread
      9. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        This is disappointing. Ad-vectored vaccines are easier than mRNA vaccines to store and distribute, and are going to be important for vaccinating people in hard-to-reach places. Despite rare side effects, they are generally considered safe and trial data shows they are effective.

        11 replies 130 retweets 953 likes
        Show this thread
      10. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        Safety concerns have already reduced enthusiasm for these vaccines, and an unforced error like this further amplifies those concerns and decreases trust in the technology itself. I hope Gamaleya resolves these issues and supports it with data and robust QC/QA.

        9 replies 123 retweets 834 likes
        Show this thread
      11. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        PS-though I wish it were for different reasons, I’m always thankful for the opportunity to practice my Russian exclamations. Да ты что, Гамалея!

        18 replies 62 retweets 680 likes
        Show this thread
      12. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        Dr. Angela Rasmussen Retweeted David Rach

        More info here: apparently the Ad5 vector was capable of forming plaques on A549 cells. The explanation is recombination, which begs the question: if it was manufactured using best practices, WTF did the vaccine vector recombine with? h/t @hildabasthttps://twitter.com/DavidRach2/status/1387183627193229313?s=20 …

        Dr. Angela Rasmussen added,

        David Rach @DavidRach2
        Replying to @DavidRach2 @Dereklowe
        Still watching the Anvisa presentation (its in Portuguese, sorry) but looks to be Ad5 vaccine lot component was forming plaques in A549 cells (https://youtu.be/loqGRfjIq8c ), method slide at 21:28. The explanation was recombinantation event. They haven't yet looked in Ad26 lots.
        12 replies 140 retweets 698 likes
        Show this thread
      13. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        Could be either a wild-type AdV or a packaging vector. To make replication-deficient vaccine vectors, you have to grow the virus in a "packaging" cell line expressing E1 in trans (on a separate piece of DNA called a plasmid).

        4 replies 66 retweets 455 likes
        Show this thread
      14. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        At this stage recombination could occur with E1 DNA from the packaging cell line, but usually there are QC measures in place to ensure this doesn't happen. Deleting E1/E3 in many Ad-vectored systems is also needed to make "room" in the genome for the insert, SARS-CoV-2, spike.

        3 replies 62 retweets 414 likes
        Show this thread
      15. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        I don't know enough about the specific AdV system used here, but it's reasonable to think that if E1 recombined w/ the Ad vector, it might replace spike. So that's doubly bad: you not only have a replication-competent Ad5, it's not expressing spike. Which is...the entire point.

        16 replies 96 retweets 553 likes
        Show this thread
      16. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        One thing is clear: if the vaccine were forming plaques on A549s, that means there's infectious, fully replication competent virus there. A plaque is a "hole" in a carpet (monolayer) of cells on a culture plate caused by cytopathic effect (virus killing cells).

        17 replies 73 retweets 480 likes
        Show this thread
      17. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        The plaque assay is a classical virological technique used to titrate infectious virus. An individual cell killed by a virus can't be seen without a microscope. To see a plaque with the naked eye, the virus has to spread to surrounding cells, and to do that it has to replicate.

        4 replies 55 retweets 442 likes
        Show this thread
      18. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        No visible plaque=no spread to surrounding cells=no viral replication. So if they saw plaques on A549s (a type of lung cell that is convenient for plaquing adenoviruses), that means there was fully replication competent virus. It's a basic and inexcusable failure of QC/QA.

        27 replies 115 retweets 670 likes
        Show this thread
      19. Dr. Angela Rasmussen‏Verified account @angie_rasmussen Apr 28

        Here's a GIF that presents an analogous situation of what the Anvisa regulators observed:pic.twitter.com/CRcSKEgeNm

        26 replies 32 retweets 394 likes
        Show this thread
      20. End of conversation

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