Multivalent Co-assembly of LL37-CpG nanoparticles: Enhanced immune response through activating multiple cell internalization pathways

Recent studies have demonstrated that the human antimicrobial peptide LL37 plays a critical role in immune regulation under both normal physiological conditions and during disease progression, as evidenced by its elevated levels observed in various chronic inflammatory diseases. A deeper understandi...

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Main Authors: Yushuang Wei, Wenwen Li, Rong Xu, Cheng Xu, Xiangyang Li, Ning Li, Fengdan Xu, Kai Yang, Bing Yuan
Format: Article
Language:English
Published: Elsevier 2025-08-01
Series:Materials Today Bio
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Online Access:http://www.sciencedirect.com/science/article/pii/S2590006425005812
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author Yushuang Wei
Wenwen Li
Rong Xu
Cheng Xu
Xiangyang Li
Ning Li
Fengdan Xu
Kai Yang
Bing Yuan
author_facet Yushuang Wei
Wenwen Li
Rong Xu
Cheng Xu
Xiangyang Li
Ning Li
Fengdan Xu
Kai Yang
Bing Yuan
author_sort Yushuang Wei
collection DOAJ
description Recent studies have demonstrated that the human antimicrobial peptide LL37 plays a critical role in immune regulation under both normal physiological conditions and during disease progression, as evidenced by its elevated levels observed in various chronic inflammatory diseases. A deeper understanding of its mechanism is essential for elucidating associated physiological and pathological processes, as well as for designing effective immune adjuvants and clinical therapeutics. In this study, we report that LL37 facilitates the assembly of unmethylated CpG dinucleotides (CpG ODNs), a clinically relevant immune adjuvant, into non-crystalline nanoparticles (NPs) with controlled size and zeta potential in a charge ratio-dependent manner. These assembled NPs enter cells via receptor-mediated macropinocytosis, coupled with LL37-mediated membrane penetration, thereby significantly enhancing cellular uptake of CpG compared to CpG alone, which enters cells through clathrin-mediated endocytosis. Consequently, the enhanced internalization of LL37-CpG NPs markedly boosts TNF-α production (>3.5-fold) in macrophages through interactions with lysosomal TLR9. This immunostimulatory mechanism offers an alternative perspective on how LL37 modulates nucleic acid assembly and activates immune cells, providing guidance for the application of peptide-CpG ODN complexes in vaccine adjuvants and immune modulation for disease treatment.
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spelling doaj-art-ad9d533ada3c4123b33badf06bc71a482025-08-20T02:20:51ZengElsevierMaterials Today Bio2590-00642025-08-013310201110.1016/j.mtbio.2025.102011Multivalent Co-assembly of LL37-CpG nanoparticles: Enhanced immune response through activating multiple cell internalization pathwaysYushuang Wei0Wenwen Li1Rong Xu2Cheng Xu3Xiangyang Li4Ning Li5Fengdan Xu6Kai Yang7Bing Yuan8Songshan Lake Materials Laboratory, Dongguan, 523808, Guangdong, China; Corresponding author.Songshan Lake Materials Laboratory, Dongguan, 523808, Guangdong, ChinaNational Laboratory of Solid State Microstructures and Department of Physics, Collaborative Innovation Center of Advanced Microstructures, Nanjing University, Nanjing, 210093, Jiangsu, China; Songshan Lake Materials Laboratory, Dongguan, 523808, Guangdong, ChinaSongshan Lake Materials Laboratory, Dongguan, 523808, Guangdong, ChinaSongshan Lake Materials Laboratory, Dongguan, 523808, Guangdong, ChinaDepartment of Neonatology, Dongguan Children's Hospital Affiliated to Guangdong Medical University, Dongguan, 523325, Guangdong, ChinaDepartment of Neonatology, Dongguan Children's Hospital Affiliated to Guangdong Medical University, Dongguan, 523325, Guangdong, ChinaCenter for Soft Condensed Matter Physics and Interdisciplinary Research & School of Physical Science and Technology, Soochow University, Suzhou, 215006, Jiangsu, China; Corresponding author.Songshan Lake Materials Laboratory, Dongguan, 523808, Guangdong, China; Corresponding author.Recent studies have demonstrated that the human antimicrobial peptide LL37 plays a critical role in immune regulation under both normal physiological conditions and during disease progression, as evidenced by its elevated levels observed in various chronic inflammatory diseases. A deeper understanding of its mechanism is essential for elucidating associated physiological and pathological processes, as well as for designing effective immune adjuvants and clinical therapeutics. In this study, we report that LL37 facilitates the assembly of unmethylated CpG dinucleotides (CpG ODNs), a clinically relevant immune adjuvant, into non-crystalline nanoparticles (NPs) with controlled size and zeta potential in a charge ratio-dependent manner. These assembled NPs enter cells via receptor-mediated macropinocytosis, coupled with LL37-mediated membrane penetration, thereby significantly enhancing cellular uptake of CpG compared to CpG alone, which enters cells through clathrin-mediated endocytosis. Consequently, the enhanced internalization of LL37-CpG NPs markedly boosts TNF-α production (>3.5-fold) in macrophages through interactions with lysosomal TLR9. This immunostimulatory mechanism offers an alternative perspective on how LL37 modulates nucleic acid assembly and activates immune cells, providing guidance for the application of peptide-CpG ODN complexes in vaccine adjuvants and immune modulation for disease treatment.http://www.sciencedirect.com/science/article/pii/S2590006425005812Single-stranded nucleic acidsSelf-assembled nanoparticleLL37Immune regulationCell internalization
spellingShingle Yushuang Wei
Wenwen Li
Rong Xu
Cheng Xu
Xiangyang Li
Ning Li
Fengdan Xu
Kai Yang
Bing Yuan
Multivalent Co-assembly of LL37-CpG nanoparticles: Enhanced immune response through activating multiple cell internalization pathways
Materials Today Bio
Single-stranded nucleic acids
Self-assembled nanoparticle
LL37
Immune regulation
Cell internalization
title Multivalent Co-assembly of LL37-CpG nanoparticles: Enhanced immune response through activating multiple cell internalization pathways
title_full Multivalent Co-assembly of LL37-CpG nanoparticles: Enhanced immune response through activating multiple cell internalization pathways
title_fullStr Multivalent Co-assembly of LL37-CpG nanoparticles: Enhanced immune response through activating multiple cell internalization pathways
title_full_unstemmed Multivalent Co-assembly of LL37-CpG nanoparticles: Enhanced immune response through activating multiple cell internalization pathways
title_short Multivalent Co-assembly of LL37-CpG nanoparticles: Enhanced immune response through activating multiple cell internalization pathways
title_sort multivalent co assembly of ll37 cpg nanoparticles enhanced immune response through activating multiple cell internalization pathways
topic Single-stranded nucleic acids
Self-assembled nanoparticle
LL37
Immune regulation
Cell internalization
url http://www.sciencedirect.com/science/article/pii/S2590006425005812
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