Cargando…
TLR4, but Neither Dectin-1 nor Dectin-2, Participates in the Mollusk Hemocyanin-Induced Proinflammatory Effects in Antigen-Presenting Cells From Mammals
Mollusk hemocyanins have biomedical uses as carriers/adjuvants and nonspecific immunostimulants with beneficial clinical outcomes by triggering the production of proinflammatory cytokines in antigen-presenting cells (APCs) and driving immune responses toward type 1 T helper (Th1) polarization. Signi...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6554540/ https://www.ncbi.nlm.nih.gov/pubmed/31214162 http://dx.doi.org/10.3389/fimmu.2019.01136 |
_version_ | 1783424985248825344 |
---|---|
author | Jiménez, José M. Salazar, Michelle L. Arancibia, Sergio Villar, Javiera Salazar, Fabián Brown, Gordon D. Lavelle, Ed C. Martínez-Pomares, Luisa Ortiz-Quintero, Jafet Lavandero, Sergio Manubens, Augusto Becker, María Inés |
author_facet | Jiménez, José M. Salazar, Michelle L. Arancibia, Sergio Villar, Javiera Salazar, Fabián Brown, Gordon D. Lavelle, Ed C. Martínez-Pomares, Luisa Ortiz-Quintero, Jafet Lavandero, Sergio Manubens, Augusto Becker, María Inés |
author_sort | Jiménez, José M. |
collection | PubMed |
description | Mollusk hemocyanins have biomedical uses as carriers/adjuvants and nonspecific immunostimulants with beneficial clinical outcomes by triggering the production of proinflammatory cytokines in antigen-presenting cells (APCs) and driving immune responses toward type 1 T helper (Th1) polarization. Significant structural features of hemocyanins as a model antigen are their glycosylation patterns. Indeed, hemocyanins have a multivalent nature as highly mannosylated antigens. We have previously shown that hemocyanins are internalized by APCs through receptor-mediated endocytosis with proteins that contain C-type lectin domains, such as mannose receptor (MR). However, the contribution of other innate immune receptors to the proinflammatory signaling pathway triggered by hemocyanins is unknown. Thus, we studied the roles of Dectin-1, Dectin-2, and Toll-like receptor 4 (TLR4) in the hemocyanin activation of murine APCs, both in dendritic cells (DCs) and macrophages, using hemocyanins from Megathura crenulata (KLH), Concholepas concholepas (CCH) and Fissurella latimarginata (FLH). The results showed that these hemocyanins bound to chimeric Dectin-1 and Dectin-2 receptors in vitro; which significantly decreased when the glycoproteins were deglycosylated. However, hemocyanin-induced proinflammatory effects in APCs from Dectin-1 knock-out (KO) and Dectin-2 KO mice were independent of both receptors. Moreover, when wild-type APCs were cultured in the presence of hemocyanins, phosphorylation of Syk kinase was not detected. We further showed that KLH and FLH induced ERK1/2 phosphorylation, a key event involved in the TLR signaling pathway. We confirmed a glycan-dependent binding of hemocyanins to chimeric TLR4 in vitro. Moreover, DCs from mice deficient for MyD88-adapter-like (Mal), a downstream adapter molecule of TLR4, were partially activated by FLH, suggesting a role of the TLR pathway in hemocyanin recognition to activate APCs. The participation of TLR4 was confirmed through a decrease in IL-12p40 and IL-6 secretion induced by FLH when a TLR4 blocking antibody was used; a reduction was also observed in DCs from C3H/HeJ mice, a mouse strain with a nonfunctional mutation for this receptor. Moreover, IL-6 secretion induced by FLH was abolished in macrophages deficient for TLR4. Our data showed the involvement of TLR4 in the hemocyanin-mediated proinflammatory response in APCs, which could cooperate with MR in innate immune recognition of these glycoproteins. |
format | Online Article Text |
id | pubmed-6554540 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-65545402019-06-18 TLR4, but Neither Dectin-1 nor Dectin-2, Participates in the Mollusk Hemocyanin-Induced Proinflammatory Effects in Antigen-Presenting Cells From Mammals Jiménez, José M. Salazar, Michelle L. Arancibia, Sergio Villar, Javiera Salazar, Fabián Brown, Gordon D. Lavelle, Ed C. Martínez-Pomares, Luisa Ortiz-Quintero, Jafet Lavandero, Sergio Manubens, Augusto Becker, María Inés Front Immunol Immunology Mollusk hemocyanins have biomedical uses as carriers/adjuvants and nonspecific immunostimulants with beneficial clinical outcomes by triggering the production of proinflammatory cytokines in antigen-presenting cells (APCs) and driving immune responses toward type 1 T helper (Th1) polarization. Significant structural features of hemocyanins as a model antigen are their glycosylation patterns. Indeed, hemocyanins have a multivalent nature as highly mannosylated antigens. We have previously shown that hemocyanins are internalized by APCs through receptor-mediated endocytosis with proteins that contain C-type lectin domains, such as mannose receptor (MR). However, the contribution of other innate immune receptors to the proinflammatory signaling pathway triggered by hemocyanins is unknown. Thus, we studied the roles of Dectin-1, Dectin-2, and Toll-like receptor 4 (TLR4) in the hemocyanin activation of murine APCs, both in dendritic cells (DCs) and macrophages, using hemocyanins from Megathura crenulata (KLH), Concholepas concholepas (CCH) and Fissurella latimarginata (FLH). The results showed that these hemocyanins bound to chimeric Dectin-1 and Dectin-2 receptors in vitro; which significantly decreased when the glycoproteins were deglycosylated. However, hemocyanin-induced proinflammatory effects in APCs from Dectin-1 knock-out (KO) and Dectin-2 KO mice were independent of both receptors. Moreover, when wild-type APCs were cultured in the presence of hemocyanins, phosphorylation of Syk kinase was not detected. We further showed that KLH and FLH induced ERK1/2 phosphorylation, a key event involved in the TLR signaling pathway. We confirmed a glycan-dependent binding of hemocyanins to chimeric TLR4 in vitro. Moreover, DCs from mice deficient for MyD88-adapter-like (Mal), a downstream adapter molecule of TLR4, were partially activated by FLH, suggesting a role of the TLR pathway in hemocyanin recognition to activate APCs. The participation of TLR4 was confirmed through a decrease in IL-12p40 and IL-6 secretion induced by FLH when a TLR4 blocking antibody was used; a reduction was also observed in DCs from C3H/HeJ mice, a mouse strain with a nonfunctional mutation for this receptor. Moreover, IL-6 secretion induced by FLH was abolished in macrophages deficient for TLR4. Our data showed the involvement of TLR4 in the hemocyanin-mediated proinflammatory response in APCs, which could cooperate with MR in innate immune recognition of these glycoproteins. Frontiers Media S.A. 2019-05-31 /pmc/articles/PMC6554540/ /pubmed/31214162 http://dx.doi.org/10.3389/fimmu.2019.01136 Text en Copyright © 2019 Jiménez, Salazar, Arancibia, Villar, Salazar, Brown, Lavelle, Martínez-Pomares, Ortiz-Quintero, Lavandero, Manubens and Becker. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Immunology Jiménez, José M. Salazar, Michelle L. Arancibia, Sergio Villar, Javiera Salazar, Fabián Brown, Gordon D. Lavelle, Ed C. Martínez-Pomares, Luisa Ortiz-Quintero, Jafet Lavandero, Sergio Manubens, Augusto Becker, María Inés TLR4, but Neither Dectin-1 nor Dectin-2, Participates in the Mollusk Hemocyanin-Induced Proinflammatory Effects in Antigen-Presenting Cells From Mammals |
title | TLR4, but Neither Dectin-1 nor Dectin-2, Participates in the Mollusk Hemocyanin-Induced Proinflammatory Effects in Antigen-Presenting Cells From Mammals |
title_full | TLR4, but Neither Dectin-1 nor Dectin-2, Participates in the Mollusk Hemocyanin-Induced Proinflammatory Effects in Antigen-Presenting Cells From Mammals |
title_fullStr | TLR4, but Neither Dectin-1 nor Dectin-2, Participates in the Mollusk Hemocyanin-Induced Proinflammatory Effects in Antigen-Presenting Cells From Mammals |
title_full_unstemmed | TLR4, but Neither Dectin-1 nor Dectin-2, Participates in the Mollusk Hemocyanin-Induced Proinflammatory Effects in Antigen-Presenting Cells From Mammals |
title_short | TLR4, but Neither Dectin-1 nor Dectin-2, Participates in the Mollusk Hemocyanin-Induced Proinflammatory Effects in Antigen-Presenting Cells From Mammals |
title_sort | tlr4, but neither dectin-1 nor dectin-2, participates in the mollusk hemocyanin-induced proinflammatory effects in antigen-presenting cells from mammals |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6554540/ https://www.ncbi.nlm.nih.gov/pubmed/31214162 http://dx.doi.org/10.3389/fimmu.2019.01136 |
work_keys_str_mv | AT jimenezjosem tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT salazarmichellel tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT arancibiasergio tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT villarjaviera tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT salazarfabian tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT browngordond tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT lavelleedc tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT martinezpomaresluisa tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT ortizquinterojafet tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT lavanderosergio tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT manubensaugusto tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals AT beckermariaines tlr4butneitherdectin1nordectin2participatesinthemolluskhemocyanininducedproinflammatoryeffectsinantigenpresentingcellsfrommammals |