title: Binding to gangliosides containing N-acetylneuraminic acid is sufficient to mediate the immunomodulatory properties of the nontoxic mucosal adjuvant LT-IIb(T13I).
abstract: By use of a mouse mucosal immunization model, LT-IIb(T13I), a nontoxic mutant type II heat-labile enterotoxin, was shown to have potent mucosal and systemic adjuvant properties. In contrast to LT-IIb, which binds strongly to ganglioside receptors decorated with either N-acetylneuraminic acid (NeuAc) or N-glycolylneuraminic acid (NeuGc), LT-IIb(T13I) binds NeuAc gangliosides much less well. Rather, LT-IIb(T13I) binds preferentially to NeuGc gangliosides. To determine if the adjuvant properties of LT-IIb(T13I) are altered in the absence of NeuGc ganglioside receptors, experiments were conducted using a Cmah-null mouse line which is deficient in the synthesis of NeuGc gangliosides. Several immunomodulatory properties of LT-IIb(T13I) were shown to be dependent on NeuGc gangliosides. LT-IIb(T13I) had reduced binding activity for NeuGc-deficient B cells and macrophages; binding to NeuGc-deficient T cells and dendritic cells (DC) was essentially undetectable. Treatment of Cmah-null macrophages with LT-IIb(T13I), however, upregulated the transcription of interleukin-4 (IL-4), IL-6, IL-17, and gamma interferon (IFN-gamma), four cytokines important for promoting immune responses. The production of mucosal IgA and serum IgG against an immunizing antigen was augmented in NeuGc-deficient mice administered LT-IIb(T13I) as a mucosal adjuvant. Notably, NeuGc gangliosides are not expressed in humans. Still, treatment of human monocytes with LT-IIb(T13I) induced the secretion of IL-6, an inflammatory cytokine that mediates differential control of leukocyte activation. These results suggested that NeuAc gangliosides are sufficient to mediate the immunomodulatory properties of LT-IIb(T13I) in mice and in human cells. The nontoxic mutant enterotoxin LT-IIb(T13I), therefore, is potentially a new and safe human mucosal adjuvant.
authors.affiliation: Witebsky Center for Microbial Pathogenesis and Immunology and Department of Microbiology and Immunology, University at Buffalo, State University of New York at Buffalo, Buffalo, New York 14214, USA.
authors.forename: Hesham F
authors.initials: HF
authors.lastname: Nawar
authors.forename: Charles S
authors.initials: CS
authors.lastname: Berenson
authors.forename: George
authors.initials: G
authors.lastname: Hajishengallis
authors.forename: Hiromu
authors.initials: H
authors.lastname: Takematsu
authors.forename: Lorrie
authors.initials: L
authors.lastname: Mandell
authors.forename: Ragina L
authors.initials: RL
authors.lastname: Clare
authors.forename: Terry D
authors.initials: TD
authors.lastname: Connell
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D001402:B-Lymphocytes / Q000737:chemistry / Q000276:immunology; D001427:Bacterial Toxins / Q000276:immunology* / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D004768:Enterotoxins / Q000276:immunology* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000276:immunology* / Q000378:metabolism*; D005732:Gangliosides / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D018928:Immunity, Mucosal; D007114:Immunization; D007155:Immunologic Factors / Q000276:immunology; D017737:Macrophages, Peritoneal / Q000737:chemistry / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D009000:Monocytes / Q000737:chemistry / Q000276:immunology; D019158:N-Acetylneuraminic Acid / Q000737:chemistry*; D011956:Receptors, Cell Surface / Q000378:metabolism
chemical_list: D000276:Adjuvants, Immunologic; D001427:Bacterial Toxins; D016207:Cytokines; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D005732:Gangliosides; D007155:Immunologic Factors; D011956:Receptors, Cell Surface; C034993:ganglioside receptor; C027538:heat-labile enterotoxin, E coli; D019158:N-Acetylneuraminic Acid
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