title: Hemocyanins Stimulate Innate Immunity by Inducing Different Temporal Patterns of Proinflammatory Cytokine Expression in Macrophages.
abstract: Hemocyanins induce a potent Th1-dominant immune response with beneficial clinical outcomes when used as a carrier/adjuvant in vaccines and nonspecific immunostimulant in cancer. However, the mechanisms by which hemocyanins trigger innate immune responses, leading to beneficial adaptive immune responses, are unknown. This response is triggered by a proinflammatory signal from various components, of which macrophages are an essential part. To understand how these proteins influence macrophage response, we investigated the effects of mollusks hemocyanins with varying structural and immunological properties, including hemocyanins from Concholepas concholepas, Fissurella latimarginata, and Megathura crenulata (keyhole limpet hemocyanin), on cultures of peritoneal macrophages. Hemocyanins were phagocytosed and slowly processed. Analysis of this process showed differential gene expression along with protein levels of proinflammatory markers, including IL-1β, IL-6, IL-12p40, and TNF-α. An extended expression analysis of 84 cytokines during a 24-h period showed a robust proinflammatory response for F. latimarginata hemocyanin in comparison with keyhole limpet hemocyanin and C. concholepas hemocyanin, which was characterized by an increase in the transcript levels of M1 cytokines involved in leukocyte recruitment. These cytokine genes included chemokines (Cxcl1, Cxcl3, Cxcl5, Ccl2, and Ccl3), ILs (Il1b and Ifng), growth factors (Csf2 and Csf3), and TNF family members (Cd40lg). The protein levels of certain cytokines were increased. However, every hemocyanin maintains downregulated key M2 cytokine genes, including Il4 and Il5 Collectively, our data demonstrate that hemocyanins are able to trigger the release of proinflammatory factors with different patterns of cytokine expression, suggesting differential signaling pathways and transcriptional network mechanisms that lead to the activation of M1-polarized macrophages.
authors.affiliation: Fundación Ciencia y Tecnología para el Desarrollo, Santiago 7750269, Chile; and.
authors.forename: Ta-Ying
authors.identifier: 0000-0001-6442-8004
authors.initials: TY
authors.lastname: Zhong
authors.affiliation: Fundación Ciencia y Tecnología para el Desarrollo, Santiago 7750269, Chile; and.
authors.forename: Sergio
authors.identifier: 0000-0002-3421-2920
authors.initials: S
authors.lastname: Arancibia
authors.affiliation: Fundación Ciencia y Tecnología para el Desarrollo, Santiago 7750269, Chile; and.
authors.forename: Raimundo
authors.initials: R
authors.lastname: Born
authors.affiliation: Fundación Ciencia y Tecnología para el Desarrollo, Santiago 7750269, Chile; and.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Tampe
authors.affiliation: Fundación Ciencia y Tecnología para el Desarrollo, Santiago 7750269, Chile; and.
authors.forename: Javiera
authors.identifier: 0000-0002-8581-4155
authors.initials: J
authors.lastname: Villar
authors.affiliation: Fundación Ciencia y Tecnología para el Desarrollo, Santiago 7750269, Chile; and.
authors.forename: Miguel
authors.identifier: 0000-0003-3300-9784
authors.initials: M
authors.lastname: Del Campo
authors.affiliation: Biosonda Corporation, Santiago 7750269, Chile.
authors.forename: Augusto
authors.identifier: 0000-0002-7931-4314
authors.initials: A
authors.lastname: Manubens
authors.affiliation: Fundación Ciencia y Tecnología para el Desarrollo, Santiago 7750269, Chile; and Biosonda Corporation, Santiago 7750269, Chile mariaines.becker@fucited.cl.
authors.forename: María Inés
authors.identifier: 0000-0002-9711-2222
authors.initials: MI
authors.lastname: Becker
mesh_terms: D000818:Animals; D016207:Cytokines / Q000235:genetics* / Q000276:immunology; D015536:Down-Regulation / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects*; D006433:Hemocyanins / Q000494:pharmacology*; D007113:Immunity, Innate / Q000187:drug effects* / Q000276:immunology; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D013997:Time Factors; D059467:Transcriptome
chemical_list: D016207:Cytokines; D006433:Hemocyanins