title: Rapid monoclonal antibody generation via dendritic cell targeting in vivo.
abstract: Dendritic cells (DC) are the professional antigen-presenting cells of the immune system. Previous studies have demonstrated that targeting foreign antigens to DC leads to enhanced antigen (Ag)-specific responses in vivo. However, the utility of this strategy for the generation of MAbs has not been investigated. To address this question we immunized mice with IgG-peptide conjugates prepared with the hamster anti-murine CD11c MAb N418. Synthetic peptides corresponding to two different exposed regions of DC-specific ICAM-3 grabbing nonintegrin (DC-SIGN), a human C-type lectin, were conjugated to N418 using thiol-based chemistry. The N418 MAb served as the targeting molecule and synthetic peptides as the Ag (MAb-Ag). A rapid and peptide specific serum IgG response was produced by Day 7 when the synthetic peptides were linked to the N418 MAb, compared to peptide co-delivered with the N418 without linkage. Spleen cells from N418-peptide immunized mice were fused on Day 10, and three IgG1/k monoclonal antibodies (MAbs) were selected to one of the peptide epitopes (MID-peptide). One of the MAbs, Novik 2, bound to two forms of recombinant DC-SIGN protein in enzyme-linked immunosorbent assay (ELISA), and was specifically inhibited by the MID-peptide in solution. Two of these MAbs show specific binding to DC-SIGN expressed by cultured human primary DC. We conclude that in vivo DC targeting enhances the immunogenicity of synthetic peptides and is an effective method for the rapid generation of MAbs to predetermined epitopes.
authors.affiliation: The Skaggs Institute for Chemical Biology, The Scripps Research Institute, BCC-515, 10550 North Torrey Pines Road, La Jolla, CA 92126, USA.
authors.forename: Jody D
authors.initials: JD
authors.lastname: Berry
authors.forename: Alexei
authors.initials: A
authors.lastname: Licea
authors.forename: Mikhail
authors.initials: M
authors.lastname: Popkov
authors.forename: Xochitl
authors.initials: X
authors.lastname: Cortez
authors.forename: Roberta
authors.initials: R
authors.lastname: Fuller
authors.forename: Marikka
authors.initials: M
authors.lastname: Elia
authors.forename: Lisa
authors.initials: L
authors.lastname: Kerwin
authors.forename: Diane
authors.initials: D
authors.lastname: Kubitz
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Barbas
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D039521:CD11c Antigen / Q000276:immunology; D015815:Cell Adhesion Molecules / Q000032:analysis / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology*; D005434:Flow Cytometry; D006801:Humans; D037181:Lectins, C-Type / Q000032:analysis / Q000276:immunology; D051379:Mice; D010455:Peptides / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000032:analysis / Q000276:immunology; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin
chemical_list: D000911:Antibodies, Monoclonal; D039521:CD11c Antigen; D015815:Cell Adhesion Molecules; D037181:Lectins, C-Type; D010455:Peptides; D011956:Receptors, Cell Surface; D000099215:DC-Specific ICAM-3 Grabbing Nonintegrin