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ABSTRACT Tolerogenic antigen-presenting cells (APCs) are promising as therapeutics for suppressing T cell activation in autoimmune diseases. However, the isolation and ex vivo manipulation

of autologous APCs is costly, and the process is customized for each patient. Here we show that tolerogenic APCs can be generated in vivo by delivering, via lipid nanoparticles, messenger

RNA coding for the inhibitory protein programmed death ligand 1. We optimized a lipid-nanoparticle formulation to minimize its immunogenicity by reducing the molar ratio of nitrogen atoms on

the ionizable lipid and the phosphate groups on the encapsulated mRNA. In mouse models of rheumatoid arthritis and ulcerative colitis, subcutaneous delivery of nanoparticles encapsulating

mRNA encoding programmed death ligand 1 reduced the fraction of activated T cells, promoted the induction of regulatory T cells and effectively prevented disease progression. The method may

allow for the engineering of APCs that target specific autoantigens or that integrate additional inhibitory molecules. Access through your institution Buy or subscribe This is a preview of

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ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS A NANOPARTICLE VACCINE THAT TARGETS

NEOANTIGEN PEPTIDES TO LYMPHOID TISSUES ELICITS ROBUST ANTITUMOR T CELL RESPONSES Article Open access 12 November 2020 A NANOVACCINE FOR ANTIGEN SELF-PRESENTATION AND IMMUNOSUPPRESSION

REVERSAL AS A PERSONALIZED CANCER IMMUNOTHERAPY STRATEGY Article 11 April 2022 BIOMIMETIC NANOVACCINE-MEDIATED MULTIVALENT IL-15 SELF-TRANSPRESENTATION (MIST) FOR POTENT AND SAFE CANCER

IMMUNOTHERAPY Article Open access 24 October 2023 DATA AVAILABILITY The data supporting the results in this study are available within the paper and its Supplementary Information. The raw

and analysed datasets generated during the study are available for research purposes from the corresponding authors on reasonable request. Source data are provided with this paper.

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_Osteoarthr. Cartil._ 14, 13–29 (2006). Google Scholar  Download references ACKNOWLEDGEMENTS This work was supported by the National Natural Science Foundation of China (52025036 to Y.W.,

82173390 to M.L. and 52495014 to Y.W.), the National Key R&D Program of China (2020YFA0710700 and 2022YFC2303300 to Y.W.), the Strategic Priority Research Program of the Chinese Academy

of Sciences (XDB0490000 and XDB0940303 to Y.W.), the Anhui Provincial Key Research and Development Project (2023s07020019 to Y.W.), the Anhui Provincial Major Science and Technology Project

(202303a07020010 to Y.W.), the Anhui Provincial Natural Science Foundation (2408085J042 to M.L.), the project of collaborative innovation for colleges of Anhui province (GXXT-2022-063 to

M.L.) and the USTC Research Funds of the Double First-Class Initiative (YD9100002054 to Y.W. and YD9110002021 to M.L.). This work was partially carried out at the USTC Center for Micro and

Nanoscale Research and Fabrication. This work was partially carried out at the Instruments Center for Physical Science, University of Science and Technology of China. AUTHOR INFORMATION

Author notes * These authors contributed equally: Yang Liu, Qian Liu, Baowen Zhang. AUTHORS AND AFFILIATIONS * Department of Radiology, the First Affiliated Hospital of University of Science

and Technology of China, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China Yang Liu, Qian Liu, Shanshan Chen, Min Li & Yucai Wang *

National Key Laboratory of Immune Response and Immunotherapy, School of Basic Medical Sciences, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei,

China Yang Liu, Qian Liu, Baowen Zhang, Yanqiong Shen, Zhibin Li, Jiachen Zhang, Min Li & Yucai Wang * Institute of Health and Medicine, Hefei Comprehensive National Science Center,

Hefei, China Yanqiong Shen & Yucai Wang * RNAlfa Biotech, Hefei, China Yanqiong Shen, Yi Yang & Yucai Wang * Key Laboratory of Anhui Province for Emerging and Reemerging Infectious

Diseases, Hefei, China Min Li & Yucai Wang Authors * Yang Liu View author publications You can also search for this author inPubMed Google Scholar * Qian Liu View author publications You

can also search for this author inPubMed Google Scholar * Baowen Zhang View author publications You can also search for this author inPubMed Google Scholar * Shanshan Chen View author

publications You can also search for this author inPubMed Google Scholar * Yanqiong Shen View author publications You can also search for this author inPubMed Google Scholar * Zhibin Li View

author publications You can also search for this author inPubMed Google Scholar * Jiachen Zhang View author publications You can also search for this author inPubMed Google Scholar * Yi

Yang View author publications You can also search for this author inPubMed Google Scholar * Min Li View author publications You can also search for this author inPubMed Google Scholar *

Yucai Wang View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS Y.W., M.L., Y.L. and Q.L. conceptualized and designed the research. Y.L., Q.L.,

B.Z., S.C., Y.S., Z.L., J.Z. and Y.Y. performed the experiments. S.C. provided help in designing LNP formulations. Y.L., Q.L. and B.Z. analysed the experimental data. Y.L., M.L., Q.L., B.Z.

and Y.W. prepared the figures and wrote the paper. Y.W. supervised the project. CORRESPONDING AUTHORS Correspondence to Min Li or Yucai Wang. ETHICS DECLARATIONS COMPETING INTERESTS The

authors declare no competing interests. PEER REVIEW PEER REVIEW INFORMATION _Nature Biomedical Engineering_ thanks Jeffrey Hubbell, Tianmeng Sun and the other, anonymous, reviewer(s) for

their contribution to the peer review of this work. Peer reviewer reports are available. ADDITIONAL INFORMATION PUBLISHER’S NOTE Springer Nature remains neutral with regard to jurisdictional

claims in published maps and institutional affiliations. EXTENDED DATA EXTENDED DATA FIG. 1 IN VIVO-PRODUCED TOL-APCS INHIBIT RA PROGRESSION. A, Statistical data of OARSI score. B, C, The

percentage of IFN-γ+ (B) and TNF-α+ (C) area per FOV. D, Representative images of CD4, CD8, and Foxp3 staining from the knee joint of one mouse in a group of four. Scale bar = 200 µm. Arrows

refer to Foxp3+ cells. E-G, Number of CD4+ (E), CD8+ (F) and Foxp3+ (G) cells per FOV. RA mice were subcutaneously treated with PBS, LNPs, or LNPs/mPDL1 (5 μg mRNA) at the lower right back.

Mice treated with iTNF-α served as the positive control group. Normal group comprises healthy mice. _n_ = 4 biologically independent mice per group for data in A-C and E-G. Data are

expressed as the mean ± s.e.m. Statistical significances were determined using one-way ANOVA with Dunnett’s post hoc test. Comparisons were performed between the LNPs/mPDL1 group and each of

the other groups. N.S. is _P_ ≥ 0.05, and significant _P_ values are displayed. Source data EXTENDED DATA FIG. 2 IN VIVO-PRODUCED TOL-APCS MEDIATE POTENT THERAPEUTIC EFFECTS IN DSS-INDUCED

UC MICE. A, Representative images of CD8, Foxp3, IFN-γ, and TNF-α staining from the colon of one mouse in a group of four. Arrows refer to Foxp3+ cells. Scale bar = 200 µm. B-E, The number

of CD8+ (B) and Foxp3+ (C) cells and the percentage of IFN-γ+ (D) and TNF-α+ (E) area per FOV. Mice were treated with PBS, LNPs, or LNPs/mPDL1 (5 μg mRNA) via subcutaneous injection at the

lower right back. Mice treated with cyclosporine served as the positive control group. Normal group comprises healthy mice. _n_ = 4 biologically independent mice per group for data in B-E.

Data are expressed as the mean ± s.e.m. Statistical significances were determined using one-way ANOVA with Dunnett’s post hoc test. Comparisons were performed between the LNPs/mPDL1 group

and each of the other groups. N.S. is _P_ ≥ 0.05, and significant _P_ values are displayed. Source data SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION Supplementary figures and tables.

REPORTING SUMMARY PEER REVIEW FILE SUPPLEMENTARY DATA Source data for the supplementary figures. SOURCE DATA SOURCE DATA FIGS. 2–7 AND EXTENDED DATA FIGS. 1 AND 2 Statistical source data.

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permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Liu, Y., Liu, Q., Zhang, B. _et al._ Generation of tolerogenic antigen-presenting cells in vivo via the delivery of mRNA encoding PDL1 within

lipid nanoparticles. _Nat. Biomed. Eng_ (2025). https://doi.org/10.1038/s41551-025-01373-0 Download citation * Received: 08 January 2024 * Accepted: 27 February 2025 * Published: 28 March

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