{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T23:07:34Z","timestamp":1740179254280,"version":"3.37.3"},"reference-count":31,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2025,1,6]],"date-time":"2025-01-06T00:00:00Z","timestamp":1736121600000},"content-version":"vor","delay-in-days":158,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"Department of Biotechnology, Government of India under Centre of Excellence","award":["BT\/COE\/34\/SP15189\/2015"],"award-info":[{"award-number":["BT\/COE\/34\/SP15189\/2015"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2024,8,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>Glycoconjugate vaccines elicit robust anti-polysaccharide Ab response by recruiting T-cell help. Multiple doses of glycoconjugate vaccine are required to induce long-lasting immunity. The characteristics of anti-polysaccharide Ab response have been reported previously. However, the effect of glycoconjugate booster immunization on anti-polysaccharide and anti\u2013carrier protein Ab repertoire remains poorly understood. In this study, we used clinically relevant pneumococcal capsular polysaccharide type 14 (PCP14) conjugated with cross-reactive material 197 (CRM197) as a model glycoconjugate Ag (PCP14-CRM197). We performed a comprehensive sequence analysis of mouse mAbs generated against PCP14 and CRM197 following immunization with one or three doses of PCP14-CRM197. Analysis of the paired Ig H and L chain transcripts revealed that anti-PCP14 Ab repertoire is extremely restricted. The reoccurrence of five replacement mutations at identical positions in anti-polysaccharide mAbs generated from different mice provided evidence for Ag-driven selection in PCP14-specific B cells. Convergent evolution was observed wherein distinct V(D)J rearrangements resulted in identical or nearly identical CDR3 in anti-PCP14 mAbs. Abs that lacked DH encoded amino acids dominated the anti-PCP14 Ab response. In contrast, anti-CRM197 Ab response was quite diverse, with fewer mutations compared with the anti-PCP14 mAbs, suggesting that conjugation of the polysaccharide to a carrier protein interferes with the development of carrier protein\u2013specific Ab responses. Our findings provide molecular insights into the maturation of Ab responses driven by booster doses of glycoconjugate. This has fundamental implications for the design of glycoconjugate vaccines, especially where the development of Ab response against the carrier protein is also crucial.<\/jats:p>","DOI":"10.4049\/immunohorizons.2400055","type":"journal-article","created":{"date-parts":[[2024,8,2]],"date-time":"2024-08-02T13:49:44Z","timestamp":1722606584000},"page":"511-526","source":"Crossref","is-referenced-by-count":0,"title":["Antigen-driven Convergent Evolution of Polysaccharide-specific \u201cDH-less\u201d B Cells in Glycoconjugate Immunized Mice"],"prefix":"10.1093","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9819-4100","authenticated-orcid":false,"given":"Sachin","family":"Kushwaha","sequence":"first","affiliation":[{"name":"Molecular Immunology Laboratory, National Institute of Immunology , Aruna Asaf Ali Marg, New Delhi ,","place":["India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-5611-4802","authenticated-orcid":false,"given":"Pratiksha","family":"Shome","sequence":"additional","affiliation":[{"name":"Molecular Immunology Laboratory, National Institute of Immunology , Aruna Asaf Ali Marg, New Delhi ,","place":["India"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4869-7105","authenticated-orcid":false,"given":"Devinder","family":"Sehgal","sequence":"additional","affiliation":[{"name":"Molecular Immunology Laboratory, National Institute of Immunology , Aruna Asaf Ali Marg, New Delhi ,","place":["India"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2024,8,2]]},"reference":[{"key":"2025010608555501600_r1","doi-asserted-by":"crossref","first-page":"eaat4615","DOI":"10.1126\/scitranslmed.aat4615","article-title":"Glycoconjugate vaccines: principles and mechanisms","volume":"10","author":"Rappuoli","year":"2018","journal-title":"Sci. Transl. Med."},{"key":"2025010608555501600_r2","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1038\/s41541-019-0110-z","article-title":"Recent advances in the production of recombinant glycoconjugate vaccines","volume":"4","author":"Kay","year":"2019","journal-title":"NPJ Vaccines"},{"key":"2025010608555501600_r3","doi-asserted-by":"crossref","first-page":"5450","DOI":"10.1128\/IAI.66.11.5450-5456.1998","article-title":"Murine immune responses to Neisseria meningitidis group C capsular polysaccharide and a thymus-dependent toxoid conjugate vaccine","volume":"66","author":"Rubinstein","year":"1998","journal-title":"Infect. Immun."},{"key":"2025010608555501600_r4","doi-asserted-by":"crossref","first-page":"2539","DOI":"10.1016\/j.carres.2003.07.008","article-title":"Immunology of bacterial polysaccharide antigens","volume":"338","author":"Weintraub","year":"2003","journal-title":"Carbohydr. Res."},{"key":"2025010608555501600_r5","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1016\/S0264-410X(00)00314-5","article-title":"Serotype of Streptococcus pneumoniae capsular polysaccharide can modify the Th1\/Th2 cytokine profile and IgG subclass response to pneumococal-CRM(197) conjugate vaccines in a murine model","volume":"19","author":"Mawas","year":"2000","journal-title":"Vaccine"},{"key":"2025010608555501600_r6","doi-asserted-by":"crossref","first-page":"1602","DOI":"10.1038\/nm.2535","article-title":"A mechanism for glycoconjugate vaccine activation of the adaptive immune system and its implications for vaccine design","volume":"17","author":"Avci","year":"2011","journal-title":"Nat. Med."},{"key":"2025010608555501600_r7","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1093\/glycob\/cww062","article-title":"Carbohydrates as T-cell antigens with implications in health and disease","volume":"26","author":"Sun","year":"2016","journal-title":"Glycobiology"},{"key":"2025010608555501600_r8","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1128\/CVI.00368-10","article-title":"Characterization of gene use and efficacy of mouse monoclonal antibodies to Streptococcus pneumoniae serotype 8","volume":"18","author":"Yano","year":"2011","journal-title":"Clin. Vaccine Immunol."},{"key":"2025010608555501600_r9","doi-asserted-by":"crossref","first-page":"1502","DOI":"10.1128\/IAI.01075-08","article-title":"Efficacy of opsonic and nonopsonic serotype 3 pneumococcal capsular polysaccharide-specific monoclonal antibodies against intranasal challenge with Streptococcus pneumoniae in mice","volume":"77","author":"Tian","year":"2009","journal-title":"Infect. Immun."},{"key":"2025010608555501600_r10","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1128\/IAI.68.1.239-246.2000","article-title":"Murine immune response to Neisseria meningitidis group C capsular polysaccharide: analysis of monoclonal antibodies generated in response to a thymus-independent antigen and a thymus-dependent toxoid conjugate vaccine","volume":"68","author":"Garc\u00eda-Ojeda","year":"2000","journal-title":"Infect. Immun."},{"key":"2025010608555501600_r11","doi-asserted-by":"crossref","first-page":"1105","DOI":"10.1084\/jem.177.4.1105","article-title":"Molecular characterization of the humoral responses to Cryptococcus neoformans infection and glucuronoxylomannan-tetanus toxoid conjugate immunization","volume":"177","author":"Mukherjee","year":"1993","journal-title":"J. Exp. Med."},{"key":"2025010608555501600_r12","doi-asserted-by":"crossref","first-page":"4469","DOI":"10.1128\/IAI.67.9.4469-4476.1999","article-title":"Molecular and idiotypic analyses of the antibody response to Cryptococcus neoformans glucuronoxylomannan-protein conjugate vaccine in autoimmune and nonautoimmune mice","volume":"67","author":"Nussbaum","year":"1999","journal-title":"Infect. Immun."},{"key":"2025010608555501600_r13","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1038\/256495a0","article-title":"Continuous cultures of fused cells secreting antibody of predefined specificity","volume":"256","author":"K\u00f6hler","year":"1975","journal-title":"Nature"},{"key":"2025010608555501600_r14","doi-asserted-by":"crossref","first-page":"5570","DOI":"10.4049\/jimmunol.0803254","article-title":"Molecular dissection of antibody responses against pneumococcal surface protein A: evidence for diverse DH-less heavy chain gene usage and avidity maturation","volume":"182","author":"Rohatgi","year":"2009","journal-title":"J. Immunol."},{"key":"2025010608555501600_r15","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.jim.2008.09.017","article-title":"Systematic design and testing of nested (RT-)PCR primers for specific amplification of mouse rearranged\/expressed immunoglobulin variable region genes from small number of B cells","volume":"339","author":"Rohatgi","year":"2008","journal-title":"J. Immunol. Methods"},{"key":"2025010608555501600_r16","doi-asserted-by":"crossref","first-page":"W503","DOI":"10.1093\/nar\/gkn316","article-title":"IMGT\/V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis","volume":"36","author":"Brochet","year":"2008","journal-title":"Nucleic Acids Res."},{"key":"2025010608555501600_r17","first-page":"695","article-title":"IMGT\/V-QUEST: IMGT standardized analysis of the immunoglobulin (IG) and T cell receptor (TR) nucleotide sequences","volume":"2011","author":"Giudicelli","year":"2011","journal-title":"Cold Spring Harb. Protoc."},{"key":"2025010608555501600_r18","doi-asserted-by":"crossref","first-page":"W34","DOI":"10.1093\/nar\/gkt382","article-title":"IgBLAST: an immunoglobulin variable domain sequence analysis tool","volume":"41","author":"Ye","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"2025010608555501600_r19","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/S0145-305X(02)00039-3","article-title":"IMGT unique numbering for immunoglobulin and T cell receptor variable domains and Ig superfamily V-like domains","volume":"27","author":"Lefranc","year":"2003","journal-title":"Dev. Comp. Immunol."},{"key":"2025010608555501600_r20","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1016\/j.vaccine.2009.09.081","article-title":"Memory B and T cell responses induced by serotype 4 Streptococcus pneumoniae vaccines: longitudinal analysis comparing responses elicited by free polysaccharide, conjugate and carrier","volume":"28","author":"Deauvieau","year":"2009","journal-title":"Vaccine"},{"key":"2025010608555501600_r21","doi-asserted-by":"crossref","first-page":"4615","DOI":"10.1128\/IAI.00472-08","article-title":"Identification of the smallest structure capable of evoking opsonophagocytic antibodies against Streptococcus pneumoniae type 14","volume":"76","author":"Safari","year":"2008","journal-title":"Infect. Immun."},{"key":"2025010608555501600_r22","doi-asserted-by":"crossref","first-page":"5107","DOI":"10.1128\/IAI.70.9.5107-5114.2002","article-title":"Immunogenicity in a mouse model of a conjugate vaccine made with a synthetic single repeating unit of type 14 pneumococcal polysaccharide coupled to CRM197","volume":"70","author":"Mawas","year":"2002","journal-title":"Infect. Immun."},{"key":"2025010608555501600_r23","doi-asserted-by":"crossref","first-page":"1491","DOI":"10.1172\/JCI91192","article-title":"T cells control the generation of nanomolar-affinity anti-glycan antibodies","volume":"127","author":"Polonskaya","year":"2017","journal-title":"J. Clin. Invest."},{"key":"2025010608555501600_r24","doi-asserted-by":"crossref","first-page":"eabj1181","DOI":"10.1126\/sciimmunol.abj1181","article-title":"Salmonella Typhi Vi capsule prime-boost vaccination induces convergent and functional antibody responses","volume":"6","author":"Dahora","year":"2021","journal-title":"Sci. Immunol."},{"key":"2025010608555501600_r25","doi-asserted-by":"crossref","first-page":"1589","DOI":"10.1038\/s41591-019-0602-4","article-title":"Polyclonal and convergent antibody response to Ebola virus vaccine rVSV-ZEBOV","volume":"25","author":"Ehrhardt","year":"2019","journal-title":"Nat. Med."},{"key":"2025010608555501600_r26","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.chom.2014.05.013","article-title":"Human responses to influenza vaccination show seroconversion signatures and convergent antibody rearrangements","volume":"16","author":"Jackson","year":"2014","journal-title":"Cell. Host. Microbe"},{"key":"2025010608555501600_r27","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1016\/j.chom.2013.05.008","article-title":"Convergent antibody signatures in human dengue","volume":"13","author":"Parameswaran","year":"2013","journal-title":"Cell Host Microbe"},{"key":"2025010608555501600_r28","doi-asserted-by":"crossref","first-page":"1201033","DOI":"10.3389\/fimmu.2023.1201033","article-title":"Convergent antibody responses are associated with broad neutralization of hepatitis C virus","volume":"14","author":"Skinner","year":"2023","journal-title":"Front. Immunol."},{"key":"2025010608555501600_r29","doi-asserted-by":"crossref","first-page":"109604","DOI":"10.1016\/j.celrep.2021.109604","article-title":"Convergent antibody responses to the SARS-CoV-2 spike protein in convalescent and vaccinated individuals","volume":"36","author":"Chen","year":"2021","journal-title":"Cell Rep."},{"key":"2025010608555501600_r30","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1084\/jem.20041577","article-title":"Direct in vivo VH to JH rearrangement violating the 12\/23 rule","volume":"201","author":"Koralov","year":"2005","journal-title":"J. Exp. Med."},{"key":"2025010608555501600_r31","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.immuni.2006.04.016","article-title":"Antibody repertoires generated by VH replacement and direct VH to JH joining","volume":"25","author":"Koralov","year":"2006","journal-title":"Immunity"}],"container-title":["ImmunoHorizons"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/immunohorizons\/article-pdf\/8\/8\/511\/60189139\/ih2400055.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/immunohorizons\/article-pdf\/8\/8\/511\/60189139\/ih2400055.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,1,6]],"date-time":"2025-01-06T08:57:57Z","timestamp":1736153877000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/immunohorizons\/article\/8\/8\/511\/7848626"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,8,1]]},"references-count":31,"journal-issue":{"issue":"8","published-print":{"date-parts":[[2024,8,1]]}},"URL":"https:\/\/doi.org\/10.4049\/immunohorizons.2400055","relation":{},"ISSN":["2573-7732"],"issn-type":[{"type":"electronic","value":"2573-7732"}],"subject":[],"published-other":{"date-parts":[[2024,8]]},"published":{"date-parts":[[2024,8,1]]}}}