title: Potent antigen-specific immune response induced by infusion of spleen cells coupled with succinimidyl-4-(N-maleimidomethyl cyclohexane)-1-carboxylate (SMCC) conjugated antigens.
abstract: In the present study, we report our recently developed new approach to inducing antigen-specific immune response. We use two nucleophilic substitution "click" chemistry processes to successfully couple protein antigens or peptides to mouse spleen cells or T cells by a heterobifunctional crosslinker, succinimidyl-4-(N-maleimidomethyl cyclohexane)-1-carboxylate (SMCC) or sulfo-SMCC. SMCC and its water-soluble analog sulfo-SMCC contain N-hydroxysuccinimide (NHS) ester and maleimide groups, which allow stable covalent conjugation of amine- and sulfhydryl-containing molecules in trans. Protein coupling to cells relies on the free sulfhydryls (thiols) on cell surfaces and the free amines on protein antigens. Although the amount of protein coupled to cells is limited due to the limited number of cell surface thiols, the injection of spleen cells coupled with antigenic proteins, such as keyhole limpet hemocyanin (KLH) or ovalbumin (OVA), induces a potent antigen-specific immune response in vivo, which is even stronger than that induced by the injection of a large dose of protein plus adjuvants. In addition, short peptides coupled to purified splenic T cells also potently elicit peptide-specific T cell proliferation in vivo after injection. Further studies show that antigen-coupled spleen cell treatment leads to augmented IFN-γ-producing T cells. Our study provides a unique antigen delivery method that efficiently distributes antigen to the entire immune system, subsequently eliciting a potent antigen-specific immune response with enhanced IFN-γ production. The findings in the present study suggest that this antigen-cell coupling strategy could be employed in immunotherapy for cancers, infectious diseases as well as immune-mediated disorders.
authors.affiliation: Department of Hematology, Xuanwu Hospital, Capital Medical University, Beijing, China.
authors.forename: Yixian
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL, USA.
authors.forename: Tyler
authors.initials: T
authors.lastname: Werbel
authors.affiliation: Department of Hematology, Xuanwu Hospital, Capital Medical University, Beijing, China.
authors.forename: Suigui
authors.initials: S
authors.lastname: Wan
authors.affiliation: Cansbio Biotechnology Company, Beijing, China.
authors.forename: Haitao
authors.initials: H
authors.lastname: Wu
authors.affiliation: Department of Neuroscience, Xuanwu Hospital, Capital Medical University, Beijing, China.
authors.forename: Yaohua
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Clare-Salzler
authors.affiliation: Department of Hematology, Xuanwu Hospital, Capital Medical University, Beijing, China; Department of Pathology, Immunology and Laboratory Medicine, University of Florida College of Medicine, Gainesville, FL, USA. Electronic address: cqx65@yahoo.com.
authors.forename: Chang-Qing
authors.initials: CQ
authors.lastname: Xia
mesh_terms: D000818:Animals; D000941:Antigens / Q000737:chemistry / Q000276:immunology*; D017690:Cell Transplantation; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000737:chemistry*; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D007371:Interferon-gamma / Q000378:metabolism; D008213:Lymphocyte Activation; D008301:Maleimides / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D013154:Spleen / Q000737:chemistry / Q000166:cytology*; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D000941:Antigens; D003432:Cross-Linking Reagents; D008301:Maleimides; C025285:N-(4-carboxycyclohexylmethyl)maleimide N-hydroxysuccinimide ester; D007371:Interferon-gamma
keywords: Antigen-cell coupling; Click chemistry; Immune response; Immunotherapy; KLH; Mice; OVA; SMCC; Th1 cells