David J Vance, Saiful Basir, Carol Lyn Piazza, Graham G Willsey, H M Emranul Haque, Jacque M Tremblay, Michael J Rudolph, Beatrice Muriuki, Lisa Cavacini, David D Weis, Charles B Shoemaker, Nicholas J Mantis
{"title":"单域抗体揭示了莱姆病疫苗抗原外表面蛋白 A(OspA)上独特的杀硼表位。","authors":"David J Vance, Saiful Basir, Carol Lyn Piazza, Graham G Willsey, H M Emranul Haque, Jacque M Tremblay, Michael J Rudolph, Beatrice Muriuki, Lisa Cavacini, David D Weis, Charles B Shoemaker, Nicholas J Mantis","doi":"10.1128/iai.00084-24","DOIUrl":null,"url":null,"abstract":"<p><p>Camelid-derived, single-domain antibodies (V<sub>H</sub>Hs) have proven to be extremely powerful tools in defining the antigenic landscape of immunologically heterogeneous surface proteins. In this report, we generated a phage-displayed V<sub>H</sub>H library directed against the candidate Lyme disease vaccine antigen, outer surface protein A (OspA). Two alpacas were immunized with recombinant OspA serotype 1 from <i>Borrelia burgdorferi sensu stricto</i> strain B31, in combination with the canine vaccine RECOMBITEK Lyme containing lipidated OspA. The phage library was subjected to two rounds of affinity enrichment (\"panning\") against recombinant OspA, yielding 21 unique V<sub>H</sub>Hs within two epitope bins, as determined through competition enzyme linked immunosorbent assays (ELISAs) with a panel of OspA-specific human monoclonal antibodies. Epitope refinement was conducted by hydrogen exchange-mass spectrometry. Six of the monovalent V<sub>H</sub>Hs were expressed as human IgG1-Fc fusion proteins and shown to have functional properties associated with protective human monoclonal antibodies, including <i>B. burgdorferi</i> agglutination, outer membrane damage, and complement-dependent borreliacidal activity. The V<sub>H</sub>Hs displayed unique reactivity profiles with the seven OspA serotypes associated with <i>B. burgdorferi</i> genospecies in the United States and Europe consistent with there being unique epitopes across OspA serotypes that should be considered when designing and evaluating multivalent Lyme disease vaccines.</p>","PeriodicalId":13541,"journal":{"name":"Infection and Immunity","volume":null,"pages":null},"PeriodicalIF":2.9000,"publicationDate":"2024-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11003225/pdf/","citationCount":"0","resultStr":"{\"title\":\"Single-domain antibodies reveal unique borrelicidal epitopes on the Lyme disease vaccine antigen, outer surface protein A (OspA).\",\"authors\":\"David J Vance, Saiful Basir, Carol Lyn Piazza, Graham G Willsey, H M Emranul Haque, Jacque M Tremblay, Michael J Rudolph, Beatrice Muriuki, Lisa Cavacini, David D Weis, Charles B Shoemaker, Nicholas J Mantis\",\"doi\":\"10.1128/iai.00084-24\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Camelid-derived, single-domain antibodies (V<sub>H</sub>Hs) have proven to be extremely powerful tools in defining the antigenic landscape of immunologically heterogeneous surface proteins. In this report, we generated a phage-displayed V<sub>H</sub>H library directed against the candidate Lyme disease vaccine antigen, outer surface protein A (OspA). Two alpacas were immunized with recombinant OspA serotype 1 from <i>Borrelia burgdorferi sensu stricto</i> strain B31, in combination with the canine vaccine RECOMBITEK Lyme containing lipidated OspA. The phage library was subjected to two rounds of affinity enrichment (\\\"panning\\\") against recombinant OspA, yielding 21 unique V<sub>H</sub>Hs within two epitope bins, as determined through competition enzyme linked immunosorbent assays (ELISAs) with a panel of OspA-specific human monoclonal antibodies. Epitope refinement was conducted by hydrogen exchange-mass spectrometry. Six of the monovalent V<sub>H</sub>Hs were expressed as human IgG1-Fc fusion proteins and shown to have functional properties associated with protective human monoclonal antibodies, including <i>B. burgdorferi</i> agglutination, outer membrane damage, and complement-dependent borreliacidal activity. The V<sub>H</sub>Hs displayed unique reactivity profiles with the seven OspA serotypes associated with <i>B. burgdorferi</i> genospecies in the United States and Europe consistent with there being unique epitopes across OspA serotypes that should be considered when designing and evaluating multivalent Lyme disease vaccines.</p>\",\"PeriodicalId\":13541,\"journal\":{\"name\":\"Infection and Immunity\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.9000,\"publicationDate\":\"2024-04-09\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11003225/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Infection and Immunity\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1128/iai.00084-24\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/3/12 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q3\",\"JCRName\":\"IMMUNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Infection and Immunity","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1128/iai.00084-24","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/3/12 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"IMMUNOLOGY","Score":null,"Total":0}
Single-domain antibodies reveal unique borrelicidal epitopes on the Lyme disease vaccine antigen, outer surface protein A (OspA).
Camelid-derived, single-domain antibodies (VHHs) have proven to be extremely powerful tools in defining the antigenic landscape of immunologically heterogeneous surface proteins. In this report, we generated a phage-displayed VHH library directed against the candidate Lyme disease vaccine antigen, outer surface protein A (OspA). Two alpacas were immunized with recombinant OspA serotype 1 from Borrelia burgdorferi sensu stricto strain B31, in combination with the canine vaccine RECOMBITEK Lyme containing lipidated OspA. The phage library was subjected to two rounds of affinity enrichment ("panning") against recombinant OspA, yielding 21 unique VHHs within two epitope bins, as determined through competition enzyme linked immunosorbent assays (ELISAs) with a panel of OspA-specific human monoclonal antibodies. Epitope refinement was conducted by hydrogen exchange-mass spectrometry. Six of the monovalent VHHs were expressed as human IgG1-Fc fusion proteins and shown to have functional properties associated with protective human monoclonal antibodies, including B. burgdorferi agglutination, outer membrane damage, and complement-dependent borreliacidal activity. The VHHs displayed unique reactivity profiles with the seven OspA serotypes associated with B. burgdorferi genospecies in the United States and Europe consistent with there being unique epitopes across OspA serotypes that should be considered when designing and evaluating multivalent Lyme disease vaccines.
期刊介绍:
Infection and Immunity (IAI) provides new insights into the interactions between bacterial, fungal and parasitic pathogens and their hosts. Specific areas of interest include mechanisms of molecular pathogenesis, virulence factors, cellular microbiology, experimental models of infection, host resistance or susceptibility, and the generation of innate and adaptive immune responses. IAI also welcomes studies of the microbiome relating to host-pathogen interactions.