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Characterizing polyclonal antibody responses via currently available methods is inherently complex and difficult. Mapping epitopes in an immune response is typically incomplete, which creates a barrier to fully understanding the humoral response to antigens and hinders rational vaccine design efforts. Here, we describe a method of characterizing polyclonal responses by using electron microscopy, and we applied this method to the immunization of rabbits with an HIV-1 envelope glycoprotein vaccine candidate, BG505 SOSIP.664. We detected known epitopes within the polyclonal sera and revealed how antibody responses evolved during the prime-boosting strategy to ultimately result in a neutralizing antibody response. We uncovered previously unidentified epitopes, including an epitope proximal to one recognized by human broadly neutralizing antibodies as well as potentially distracting non-neutralizing epitopes. Our method provides an efficient and semiquantitative map of epitopes that are targeted in a polyclonal antibody response and should be of widespread utility in vaccine and infection studies.
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http://dx.doi.org/10.1016/j.immuni.2018.07.009 | DOI Listing |
Front Immunol
September 2025
Division of Allergy and Clinical Immunology, Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, United States.
NSG-SGM3 humanized mouse models are well-suited for studying human immune physiology but are technically challenging and expensive. We previously characterized a simplified NSG-SGM3 mouse, engrafted with human donor CD34 hematopoietic stem cells without receiving prior bone marrow ablation or human secondary lymphoid tissue implantation, that still retains human mast cell- and basophil-dependent passive anaphylaxis responses. Its capacities for human antibody production and human B cell maturation, however, remain unknown.
View Article and Find Full Text PDFMicrob Genom
September 2025
Clinical Microbiology and Virology Unit, 'Spirito Santo' Hospital, Via Fonte Romana n. 8, 65124 Pescara, Italy.
This study investigated an increase in bacteraemia cases amongst hospitalized patients in Italy during the summer of 2023. To precisely characterize the outbreak, we performed extensive genomic investigation, including both short- and long-read sequencing technologies, combined with bioinformatics analysis. This genomic approach enabled us to identify the putative source of the outbreak and understand the transmission dynamics of this opportunistic pathogen within the hospital.
View Article and Find Full Text PDFBioelectrochemistry
August 2025
Department of Physical Chemistry, Institute of Chemistry, Faculty of Chemistry and Geosciences, Vilnius University, Naugarduko St. 24, LT-03225 Vilnius, Lithuania; Department of Nanotechnology, State Research Institute Center for Physical and Technological Sciences, Sauletekio Ave. 3, LT-10257 Vilni
The emergence of SARS-CoV-2 has posed significant global health challenges. The nucleocapsid protein (N-protein) is a structural part of the SARS-CoV-2 virus and an important immunogenic target of specific antibodies, which are developed in the organism during the infection by this virus. Artificially designed specific (monoclonal and polyclonal) antibodies are also used for therapeutic and bioanalytical purposes, therefore, the assessment and characterization of newly designed antibodies is an important analytical issue.
View Article and Find Full Text PDFN Engl J Med
September 2025
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, ON, Canada.
Background: Heparin-induced thrombocytopenia (HIT) is an immune-mediated platelet disorder caused by antibodies that target complexes of platelet factor 4 (PF4) and heparin. HIT has been characterized as a polyclonal immune response; however, studies of other rare anti-PF4 disorders have identified clonally restricted antibodies.
Methods: In this study, we investigated the clonality of pathogenic HIT antibodies.
Brain Behav
September 2025
Department of Transplant and Infection Immunology, Saarland University, Homburg, Germany.
Background: Immune induction under B-cell depletion is complex and far from being fully understood.
Methods: We investigated clinical and immunological responses after dual homologous mRNA vaccination with BNT162b2 and after booster vaccination or infection in 14 B-cell depleted patients with inflammatory central nervous system disease in comparison to 28 healthy controls. Spike-specific IgG were determined using ELISA and neutralizing activity by surrogate assay.