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As BB0405 is highly conserved across different <jats:italic>B. burgdorferi<\/jats:italic> sensu lato species, we investigated whether vaccination with recombinant BB0405 or through intradermal <jats:italic>bb0405<\/jats:italic> DNA tattoo vaccination could provide protection against different <jats:italic>Borrelia<\/jats:italic> species, specifically against <jats:italic>Borrelia afzelii<\/jats:italic>, the predominant <jats:italic>B. burgdorferi<\/jats:italic> sensu lato genospecies causing Lyme borreliosis across Eurasia. We immunized C3H\/HeN mice with recombinant BB0405 or with a codon-optimized <jats:italic>bb0405<\/jats:italic> DNA vaccine using the pVAC plasmid and immunized corresponding control groups mice with only adjuvant or empty vectors. We subsequently subjected these immunized mice to a tick challenge with <jats:italic>B. afzelii<\/jats:italic> CB43-infected <jats:italic>Ixodes ricinus<\/jats:italic> nymphs. Upon vaccination, recombinant BB0405 induced a high total IgG response, but <jats:italic>bb0405<\/jats:italic> DNA vaccination did not elicit antibody responses. Both vaccine formulations did not provide protection against <jats:italic>Borrelia afzelii<\/jats:italic> strain CB43 after tick challenge. In an attempt to understand the lack of protection of the recombinant vaccine, we determined expression of BB0405 and showed that <jats:italic>B. afzelii<\/jats:italic> CB43 spirochetes significantly and drastically downregulate the expression of BB0405 protein at 37\u00a0\u00b0C compared to 33\u00a0\u00b0C, where as in <jats:italic>B. burgdorferi<\/jats:italic> B31 spirochetes expression levels remain unaltered. Vaccination with recombinant BB0405 was previously shown to protect against <jats:italic>B. burgdorferi<\/jats:italic> sensu stricto. Here we show that vaccination with either recombinant BB0405 (or non-immunogenic <jats:italic>bb0405<\/jats:italic> DNA), despite being highly conserved among <jats:italic>B. burgdorferi<\/jats:italic> sl genospecies, does not provide cross-protection against <jats:italic>B. afzelii<\/jats:italic>, mostly likely due to downregulation of this protein in <jats:italic>B. afzelii<\/jats:italic> in the mammalian host.<\/jats:p>","DOI":"10.1038\/s41598-021-84130-y","type":"journal-article","created":{"date-parts":[[2021,2,26]],"date-time":"2021-02-26T11:03:35Z","timestamp":1614337415000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Investigating BB0405 as a novel Borrelia afzelii vaccination candidate in Lyme borreliosis"],"prefix":"10.1038","volume":"11","author":[{"given":"M. J.","family":"Klouwens","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. 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