Proinflammatory Action of a New Electronegative Low-Density Lipoprotein Epitope
The electronegative low-density lipoprotein, LDL (−), is an endogenously modified LDL subfraction with cytotoxic and proinflammatory actions on endothelial cells, monocytes, and macrophages contributing to the progression of atherosclerosis. In this study, epitopes of LDL (−) wer...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2019-08-01
|
| Series: | Biomolecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-273X/9/8/386 |
| _version_ | 1811196167801798656 |
|---|---|
| author | Tanize do Espirito Santo Faulin Soraya Megumi Kazuma Gustavo Luis Tripodi Marcela Frota Cavalcante Felipe Wakasuqui Cristiano Luis Pinto Oliveira Maximilia Frazão de Souza Degenhardt Jussara Michaloski Ricardo José Giordano Daniel Francisco Jacon Ketelhuth Dulcineia Saes Parra Abdalla |
| author_facet | Tanize do Espirito Santo Faulin Soraya Megumi Kazuma Gustavo Luis Tripodi Marcela Frota Cavalcante Felipe Wakasuqui Cristiano Luis Pinto Oliveira Maximilia Frazão de Souza Degenhardt Jussara Michaloski Ricardo José Giordano Daniel Francisco Jacon Ketelhuth Dulcineia Saes Parra Abdalla |
| author_sort | Tanize do Espirito Santo Faulin |
| collection | DOAJ |
| description | The electronegative low-density lipoprotein, LDL (−), is an endogenously modified LDL subfraction with cytotoxic and proinflammatory actions on endothelial cells, monocytes, and macrophages contributing to the progression of atherosclerosis. In this study, epitopes of LDL (−) were mapped using a phage display library of peptides and monoclonal antibodies reactive to this modified lipoprotein. Two different peptide libraries (X6 and CX8C for 6- and 8-amino acid-long peptides, respectively) were used in the mapping. Among all tested peptides, two circular peptides, P1A3 and P2C7, were selected based on their high affinities for the monoclonal antibodies. Small-angle X-ray scattering analysis confirmed their structures as circular rings. P1A3 or P2C7 were quickly internalized by bone marrow-derived murine macrophages as shown by confocal microscopy. P2C7 increased the expression of TNFα, IL-1 β and iNOS as well as the secretion of TNFα, CCL2, and nitric oxide by murine macrophages, similar to the responses induced by LDL (−), although less intense. In contrast, P1A3 did not show pro-inflammatory effects. We identified a mimetic epitope associated with LDL (−), the P2C7 circular peptide, that activates macrophages. Our data suggest that this conformational epitope represents an important danger-associated molecular pattern of LDL (−) that triggers proinflammatory responses. |
| first_indexed | 2024-04-12T00:54:02Z |
| format | Article |
| id | doaj.art-d964ef749d23465abc868a8bac587def |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-04-12T00:54:02Z |
| publishDate | 2019-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-d964ef749d23465abc868a8bac587def2022-12-22T03:54:39ZengMDPI AGBiomolecules2218-273X2019-08-019838610.3390/biom9080386biom9080386Proinflammatory Action of a New Electronegative Low-Density Lipoprotein EpitopeTanize do Espirito Santo Faulin0Soraya Megumi Kazuma1Gustavo Luis Tripodi2Marcela Frota Cavalcante3Felipe Wakasuqui4Cristiano Luis Pinto Oliveira5Maximilia Frazão de Souza Degenhardt6Jussara Michaloski7Ricardo José Giordano8Daniel Francisco Jacon Ketelhuth9Dulcineia Saes Parra Abdalla10Department of Clinical and Toxicological Analyses, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo 05508-000, BrazilDepartment of Clinical and Toxicological Analyses, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo 05508-000, BrazilDepartment of Clinical and Toxicological Analyses, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo 05508-000, BrazilDepartment of Clinical and Toxicological Analyses, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo 05508-000, BrazilDepartment of Clinical and Toxicological Analyses, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo 05508-000, BrazilDepartment of Experimental Physics, Institute of Physics, University of Sao Paulo, Sao Paulo 05508-090, BrazilDepartment of Experimental Physics, Institute of Physics, University of Sao Paulo, Sao Paulo 05508-090, BrazilDepartment of Biochemistry, Institute of Chemistry, University of Sao Paulo, Sao Paulo 05508-000, SP, BrazilDepartment of Biochemistry, Institute of Chemistry, University of Sao Paulo, Sao Paulo 05508-000, SP, BrazilCentre for Molecular Medicine, Department of Medicine, Karolinska University Hospital, Karolinska Institute, 17164 Stockholm, SwedenDepartment of Clinical and Toxicological Analyses, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo 05508-000, BrazilThe electronegative low-density lipoprotein, LDL (−), is an endogenously modified LDL subfraction with cytotoxic and proinflammatory actions on endothelial cells, monocytes, and macrophages contributing to the progression of atherosclerosis. In this study, epitopes of LDL (−) were mapped using a phage display library of peptides and monoclonal antibodies reactive to this modified lipoprotein. Two different peptide libraries (X6 and CX8C for 6- and 8-amino acid-long peptides, respectively) were used in the mapping. Among all tested peptides, two circular peptides, P1A3 and P2C7, were selected based on their high affinities for the monoclonal antibodies. Small-angle X-ray scattering analysis confirmed their structures as circular rings. P1A3 or P2C7 were quickly internalized by bone marrow-derived murine macrophages as shown by confocal microscopy. P2C7 increased the expression of TNFα, IL-1 β and iNOS as well as the secretion of TNFα, CCL2, and nitric oxide by murine macrophages, similar to the responses induced by LDL (−), although less intense. In contrast, P1A3 did not show pro-inflammatory effects. We identified a mimetic epitope associated with LDL (−), the P2C7 circular peptide, that activates macrophages. Our data suggest that this conformational epitope represents an important danger-associated molecular pattern of LDL (−) that triggers proinflammatory responses.https://www.mdpi.com/2218-273X/9/8/386atherosclerosismacrophagesLDLoxidized LDLelectronegative LDLdanger-associated molecular patternmimotope peptidephage displayepitope mapping |
| spellingShingle | Tanize do Espirito Santo Faulin Soraya Megumi Kazuma Gustavo Luis Tripodi Marcela Frota Cavalcante Felipe Wakasuqui Cristiano Luis Pinto Oliveira Maximilia Frazão de Souza Degenhardt Jussara Michaloski Ricardo José Giordano Daniel Francisco Jacon Ketelhuth Dulcineia Saes Parra Abdalla Proinflammatory Action of a New Electronegative Low-Density Lipoprotein Epitope Biomolecules atherosclerosis macrophages LDL oxidized LDL electronegative LDL danger-associated molecular pattern mimotope peptide phage display epitope mapping |
| title | Proinflammatory Action of a New Electronegative Low-Density Lipoprotein Epitope |
| title_full | Proinflammatory Action of a New Electronegative Low-Density Lipoprotein Epitope |
| title_fullStr | Proinflammatory Action of a New Electronegative Low-Density Lipoprotein Epitope |
| title_full_unstemmed | Proinflammatory Action of a New Electronegative Low-Density Lipoprotein Epitope |
| title_short | Proinflammatory Action of a New Electronegative Low-Density Lipoprotein Epitope |
| title_sort | proinflammatory action of a new electronegative low density lipoprotein epitope |
| topic | atherosclerosis macrophages LDL oxidized LDL electronegative LDL danger-associated molecular pattern mimotope peptide phage display epitope mapping |
| url | https://www.mdpi.com/2218-273X/9/8/386 |
| work_keys_str_mv | AT tanizedoespiritosantofaulin proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT sorayamegumikazuma proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT gustavoluistripodi proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT marcelafrotacavalcante proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT felipewakasuqui proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT cristianoluispintooliveira proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT maximiliafrazaodesouzadegenhardt proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT jussaramichaloski proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT ricardojosegiordano proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT danielfranciscojaconketelhuth proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope AT dulcineiasaesparraabdalla proinflammatoryactionofanewelectronegativelowdensitylipoproteinepitope |