{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,17]],"date-time":"2026-10-17T02:54:21Z","timestamp":1792205661954,"version":"4.4.0"},"reference-count":49,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,8,22]],"date-time":"2023-08-22T00:00:00Z","timestamp":1692662400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100019707","name":"Connecticut Children's Medical Center","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100019707","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Syphilis, a sexually transmitted infection caused by the spirochete\n                      <jats:italic>Treponema pallidum<\/jats:italic>\n                      (\n                      <jats:italic>Tp<\/jats:italic>\n                      ), is resurging globally.\n                      <jats:italic>Tp<\/jats:italic>\n                      \u2019s repertoire of outer membrane proteins (OMPs) includes BamA (\u03b2-barrel assembly machinery subunit A\/TP0326), a bipartite protein consisting of a 16-stranded \u03b2-barrel with nine extracellular loops (ECLs) and five periplasmic POTRA (polypeptide transport-associated) domains. BamA ECL4 antisera promotes internalization of\n                      <jats:italic>Tp<\/jats:italic>\n                      by rabbit peritoneal macrophages.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Three overlapping BamA ECL4 peptides and a two-stage, phage display strategy, termed \u201cEpivolve\u201d (for epitope evolution) were employed to generate single-chain variable fragments (scFvs). Additionally, antisera generated by immunizing mice and rabbits with BamA ECL4 displayed by a\n                      <jats:italic>Pyrococcus furiosus<\/jats:italic>\n                      thioredoxin scaffold (\n                      <jats:italic>Pf<\/jats:italic>\n                      Trx\n                      <jats:sup>BamA\/ECL4<\/jats:sup>\n                      ). MAbs and antisera reactivities were evaluated by immunoblotting and ELISA. A comparison of murine and rabbit opsonophagocytosis assays was conducted to evaluate the functional ability of the Abs (\n                      <jats:italic>e.g.<\/jats:italic>\n                      , opsonization) and validate the mouse assay. Sera from\n                      <jats:italic>Tp<\/jats:italic>\n                      -infected mice (MSS) and rabbits (IRS) were evaluated for ECL4-specific Abs using\n                      <jats:italic>Pf<\/jats:italic>\n                      Trx\n                      <jats:sup>BamA\/ECL4<\/jats:sup>\n                      and overlapping ECL4 peptides in immunoblotting and ELISA assays.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Each of the five mAbs demonstrated reactivity by immunoblotting and ELISA to nanogram amounts of\n                      <jats:italic>Pf<\/jats:italic>\n                      Trx\n                      <jats:sup>BamA\/ECL4<\/jats:sup>\n                      . One mAb, containing a unique amino acid sequence in both the light and heavy chains, showed activity in the murine opsonophagocytosis assay. Mice and rabbits hyperimmunized with\n                      <jats:italic>Pf<\/jats:italic>\n                      Trx\n                      <jats:sup>BamA\/ECL4<\/jats:sup>\n                      produced opsonic antisera that strongly recognized the ECL presented in a heterologous scaffold and overlapping ECL4 peptides, including S2. In contrast, Abs generated during\n                      <jats:italic>Tp<\/jats:italic>\n                      infection of mice and rabbits poorly recognized the peptides, indicating that S2 contains a subdominant epitope.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      Epivolve produced mAbs target subdominant opsonic epitopes in BamA ECL4, a top syphilis vaccine candidate. The murine opsonophagocytosis assay can serve as an alternative model to investigate the opsonic potential of vaccinogens. Detailed characterization of BamA ECL4-specific Abs provided a means to dissect Ab responses elicited by\n                      <jats:italic>Tp<\/jats:italic>\n                      infection.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fimmu.2023.1222267","type":"journal-article","created":{"date-parts":[[2023,8,22]],"date-time":"2023-08-22T09:56:19Z","timestamp":1692698179000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":24,"title":["Use of Epivolve phage display to generate a monoclonal antibody with opsonic activity directed against a subdominant epitope on extracellular loop 4 of Treponema pallidum BamA (TP0326)"],"prefix":"10.3389","volume":"14","author":[{"given":"Mary R.","family":"Ferguson","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristina N.","family":"Delgado","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shannon","family":"McBride","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabel C.","family":"Orbe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carson J.","family":"La Vake","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa J.","family":"Caimano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiana","family":"Mendez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Trevor F.","family":"Moraes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anthony B.","family":"Schryvers","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M. 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