Improved acid-driven inhibition of effector T cell function by a pHLIP variant-conjugated PD-L1

Abstract Programmed cell death–ligand 1 (PD-L1)/PD-1 axis is crucial for maintenance of immune homeostasis and its impairment partially accounts for the pathogenesis of inflammatory diseases. Hence, augmenting PD-L1/PD-1 signals represents a novel strategy to prevent destructive inflammation and ind...

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Main Authors: Hang Zheng, Mianjing Wang, Junjuan Feng, Yuting Zhang, Haiyan Wu, Min Zhang, He Xiao, Chunxia Qiao, Jing Wang, Longlong Luo, Xinying Li, Jiannan Feng, Yuanqiang Zheng, Yi Wang, Dongsheng Sheng, Guojiang Chen
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Language:English
Published: Nature Portfolio 2025-04-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-98135-4
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author Hang Zheng
Mianjing Wang
Junjuan Feng
Yuting Zhang
Haiyan Wu
Min Zhang
He Xiao
Chunxia Qiao
Jing Wang
Longlong Luo
Xinying Li
Jiannan Feng
Yuanqiang Zheng
Yi Wang
Dongsheng Sheng
Guojiang Chen
author_facet Hang Zheng
Mianjing Wang
Junjuan Feng
Yuting Zhang
Haiyan Wu
Min Zhang
He Xiao
Chunxia Qiao
Jing Wang
Longlong Luo
Xinying Li
Jiannan Feng
Yuanqiang Zheng
Yi Wang
Dongsheng Sheng
Guojiang Chen
author_sort Hang Zheng
collection DOAJ
description Abstract Programmed cell death–ligand 1 (PD-L1)/PD-1 axis is crucial for maintenance of immune homeostasis and its impairment partially accounts for the pathogenesis of inflammatory diseases. Hence, augmenting PD-L1/PD-1 signals represents a novel strategy to prevent destructive inflammation and induce immune tolerance. Recently, we developed a new cargo by conjugating the ectodomain of PD-L1 with pHLIP, a low pH-responding and membrane-inserting peptide, and demonstrated its potent immune-suppressive activity under weakly acidic (pH6.1) conditions in vitro. Herein, we further showed that PD-L1-pHLIP (termed as PD-L1-pHLIPwt) responded well to weakly acidic buffer, but not in nearly neutral pH (pH6.8) solutions. To overcome this obstacle, pHLIPwt was replaced by a variant harboring two mutations (Asp14Gla and Asp25Aad) and PD-L1 ectodomain was conjugated to the N-terminus of pHLIP variant via sulfo-SMCC linker (termed as PD-L1-pHLIPva). PD-L1-pHLIPva potently inhibited T effector function including proliferation, activation as well as proinflammatory cytokine release in nearly neutral pH buffer through PD-L1/PD-1 interaction. The inhibitory function of PD-L1-pHLIPva was attributed to more amounts of PD-L1 anchored on the surface of several types of immune cells compared with PD-L1-pHLIPwt. Given that the niche in the lesions of inflammation is weakly acidic even nearly neutral pH, PD-L1-pHLIPva represents a new arsenal to potentially dampen excessive inflammatory reactions.
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spelling doaj-art-3a47a8b68f384772ac5d78cc1e75abbd2025-08-20T03:18:42ZengNature PortfolioScientific Reports2045-23222025-04-0115111410.1038/s41598-025-98135-4Improved acid-driven inhibition of effector T cell function by a pHLIP variant-conjugated PD-L1Hang Zheng0Mianjing Wang1Junjuan Feng2Yuting Zhang3Haiyan Wu4Min Zhang5He Xiao6Chunxia Qiao7Jing Wang8Longlong Luo9Xinying Li10Jiannan Feng11Yuanqiang Zheng12Yi Wang13Dongsheng Sheng14Guojiang Chen15Research Center of Molecular Biology, School of Basic Medical Sciences, Inner Mongolia Medical UniversityResearch Center of Molecular Biology, School of Basic Medical Sciences, Inner Mongolia Medical UniversityResearch Center of Molecular Biology, School of Basic Medical Sciences, Inner Mongolia Medical UniversityInstitute of Pharmacology and ToxicologyInstitute of Pharmacology and ToxicologyInstitute of Pharmacology and ToxicologyInstitute of Pharmacology and ToxicologyInstitute of Pharmacology and ToxicologyInstitute of Pharmacology and ToxicologyInstitute of Pharmacology and ToxicologyInstitute of Pharmacology and ToxicologyInstitute of Pharmacology and ToxicologyResearch Center of Molecular Biology, School of Basic Medical Sciences, Inner Mongolia Medical UniversityDepartment of Hematology, The Fifth Medical Center, Chinese PLA General HospitalDepartment of Thoracic surgery, The Fifth Medical Center, Chinese PLA General HospitalInstitute of Pharmacology and ToxicologyAbstract Programmed cell death–ligand 1 (PD-L1)/PD-1 axis is crucial for maintenance of immune homeostasis and its impairment partially accounts for the pathogenesis of inflammatory diseases. Hence, augmenting PD-L1/PD-1 signals represents a novel strategy to prevent destructive inflammation and induce immune tolerance. Recently, we developed a new cargo by conjugating the ectodomain of PD-L1 with pHLIP, a low pH-responding and membrane-inserting peptide, and demonstrated its potent immune-suppressive activity under weakly acidic (pH6.1) conditions in vitro. Herein, we further showed that PD-L1-pHLIP (termed as PD-L1-pHLIPwt) responded well to weakly acidic buffer, but not in nearly neutral pH (pH6.8) solutions. To overcome this obstacle, pHLIPwt was replaced by a variant harboring two mutations (Asp14Gla and Asp25Aad) and PD-L1 ectodomain was conjugated to the N-terminus of pHLIP variant via sulfo-SMCC linker (termed as PD-L1-pHLIPva). PD-L1-pHLIPva potently inhibited T effector function including proliferation, activation as well as proinflammatory cytokine release in nearly neutral pH buffer through PD-L1/PD-1 interaction. The inhibitory function of PD-L1-pHLIPva was attributed to more amounts of PD-L1 anchored on the surface of several types of immune cells compared with PD-L1-pHLIPwt. Given that the niche in the lesions of inflammation is weakly acidic even nearly neutral pH, PD-L1-pHLIPva represents a new arsenal to potentially dampen excessive inflammatory reactions.https://doi.org/10.1038/s41598-025-98135-4PD-L1pHLIPvaAcidityT cellImmunosuppression
spellingShingle Hang Zheng
Mianjing Wang
Junjuan Feng
Yuting Zhang
Haiyan Wu
Min Zhang
He Xiao
Chunxia Qiao
Jing Wang
Longlong Luo
Xinying Li
Jiannan Feng
Yuanqiang Zheng
Yi Wang
Dongsheng Sheng
Guojiang Chen
Improved acid-driven inhibition of effector T cell function by a pHLIP variant-conjugated PD-L1
Scientific Reports
PD-L1
pHLIPva
Acidity
T cell
Immunosuppression
title Improved acid-driven inhibition of effector T cell function by a pHLIP variant-conjugated PD-L1
title_full Improved acid-driven inhibition of effector T cell function by a pHLIP variant-conjugated PD-L1
title_fullStr Improved acid-driven inhibition of effector T cell function by a pHLIP variant-conjugated PD-L1
title_full_unstemmed Improved acid-driven inhibition of effector T cell function by a pHLIP variant-conjugated PD-L1
title_short Improved acid-driven inhibition of effector T cell function by a pHLIP variant-conjugated PD-L1
title_sort improved acid driven inhibition of effector t cell function by a phlip variant conjugated pd l1
topic PD-L1
pHLIPva
Acidity
T cell
Immunosuppression
url https://doi.org/10.1038/s41598-025-98135-4
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