title: Antibody-driven design of a human cytomegalovirus gHgLpUL128L subunit vaccine that selectively elicits potent neutralizing antibodies.
abstract: The use of neutralizing antibodies to identify the most effective antigen has been proposed as a strategy to design vaccines capable of eliciting protective B-cell immunity. In this study, we analyzed the human antibody response to cytomegalovirus (human cytomegalovirus, HCMV) infection and found that antibodies to glycoprotein (g)B, a surface glycoprotein that has been developed as a HCMV vaccine, were primarily nonneutralizing. In contrast, most of the antibodies to the complex formed by gH, gL, protein (p)UL128, pUL130, and pUL131 (the gHgLpUL128L pentamer) neutralized HCMV infection with high potency. Based on this analysis, we developed a single polycistronic vector encoding the five pentamer genes separated by "self-cleaving" 2A peptides to generate a stably transfected CHO cell line constitutively secreting high levels of recombinant pentamer that displayed the functional antigenic sites targeted by human neutralizing antibodies. Immunization of mice with the pentamer formulated with different adjuvants elicited HCMV neutralizing antibody titers that persisted to high levels over time and that were a hundred- to thousand-fold higher than those found in individuals that recovered from primary HCMV infection. Sera from mice immunized with the pentamer vaccine neutralized infection of both epithelial cells and fibroblasts and prevented cell-to-cell spread and viral dissemination from endothelial cells to leukocytes. Neutralizing monoclonal antibodies from immunized mice showed the same potency as human antibodies and targeted the same as well as additional sites on the pentamer. These results illustrate with a relevant example a general and practical approach of analytic vaccinology for the development of subunit vaccines against complex pathogens.
authors.affiliation: Institute for Research in Biomedicine, Università della Svizzera Italiana, 6500 Bellinzona, Switzerland;
authors.forename: Anna
authors.initials: A
authors.lastname: Kabanova
authors.affiliation: Institute for Research in Biomedicine, Università della Svizzera Italiana, 6500 Bellinzona, Switzerland;
authors.forename: Laurent
authors.initials: L
authors.lastname: Perez
authors.affiliation: Institute for Research in Biomedicine, Università della Svizzera Italiana, 6500 Bellinzona, Switzerland; Laboratori Sperimentali di Ricerca and.
authors.forename: Daniele
authors.initials: D
authors.lastname: Lilleri
authors.affiliation: Institute for Research in Biomedicine, Università della Svizzera Italiana, 6500 Bellinzona, Switzerland;
authors.forename: Jessica
authors.initials: J
authors.lastname: Marcandalli
authors.affiliation: Humabs BioMed SA, 6500 Bellinzona, Switzerland; and.
authors.forename: Gloria
authors.initials: G
authors.lastname: Agatic
authors.affiliation: Institute for Research in Biomedicine, Università della Svizzera Italiana, 6500 Bellinzona, Switzerland;
authors.forename: Simone
authors.initials: S
authors.lastname: Becattini
authors.affiliation: Institute for Research in Biomedicine, Università della Svizzera Italiana, 6500 Bellinzona, Switzerland;
authors.forename: Silvia
authors.initials: S
authors.lastname: Preite
authors.affiliation: Institute for Research in Biomedicine, Università della Svizzera Italiana, 6500 Bellinzona, Switzerland;
authors.forename: Dario
authors.initials: D
authors.lastname: Fuschillo
authors.affiliation: Virologia e Microbiologia, Fondazione Istituto di Ricovero e Cura a Carattere Scientifico Policlinico San Matteo, 27100 Pavia, Italy;
authors.forename: Elena
authors.initials: E
authors.lastname: Percivalle
authors.affiliation: Institute for Research in Biomedicine, Università della Svizzera Italiana, 6500 Bellinzona, Switzerland;
authors.forename: Federica
authors.initials: F
authors.lastname: Sallusto
authors.affiliation: Laboratori Sperimentali di Ricerca and.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Gerna
authors.affiliation: Humabs BioMed SA, 6500 Bellinzona, Switzerland; and.
authors.forename: Davide
authors.initials: D
authors.lastname: Corti
authors.affiliation: Institute for Research in Biomedicine, Università della Svizzera Italiana, 6500 Bellinzona, Switzerland; Institute of Microbiology, ETH Zurich, 8093 Zurich, Switzerland lanzavecchia@irb.usi.ch.
authors.forename: Antonio
authors.initials: A
authors.lastname: Lanzavecchia
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology* / Q000302:isolation & purification; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D022482:Cytomegalovirus Vaccines / Q000276:immunology*; D015195:Drug Design; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D018709:Statistics, Nonparametric; D022223:Vaccines, Subunit / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000956:Antigens, Viral; D022482:Cytomegalovirus Vaccines; D022223:Vaccines, Subunit
keywords: analytic vaccinology; human cytomegalovirus; vaccine