Abstract
Yellow fever virus (YFV) is an arbovirus causing substantial human morbidity and mortality. The live-attenuated 17D strain serves as vaccine and is one of the most successful vaccines so far. Receptor usage between attenuated and pathogenic YFV strains remains unclear. Here we performed a barcoded, genome-wide human open-reading frame library screen and identified LRP8 (also named APOER2) as a receptor for YFV. We show that LRP8 expression increases YFV infection (17D and two clinical strains, BJ01 and Asibi) in cell lines by promoting entry. Adeno-associated virus-mediated expression of human LRP8 in mouse liver aggravates infection and pathology of the clinical strain BJ01. LRP8 knockdown decreases YFV infection in brain cells, primary human hepatocytes and mosquitoes. LRP8 directly interacts with YFV 17D particles via the viral envelope protein. A soluble LRP8 decoy protein can block YFV 17D and BJ01 infection. Our findings provide insights for understanding YFV entry, tropism and pathogenesis.
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Data availability
scRNA-seq raw data have been deposited in the Genome Sequence Archive (Genomics, Proteomics and Bioinformatics 2021) in the National Genomics Data Center, China National Center for Bioinformation/Beijing Institute of Genomics, Chinese Academy of Sciences (GSA-Human: HRA013453). Source data are provided with this paper.
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Acknowledgements
This work was supported by a grant from the State Key Research Development Program of China to X.T. (grant nos. 2021YFA1302503 and 2022YFE0102200), a grant from Guangdong Basic and Applied Basic Research Foundation to Y.Y. (no. 2025B1515020010), a grant from the National Natural Science Foundation of China to Z.W. (82502710) and a grant from the National Excellent Young Physician Project from the National Health Commission to X.H. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. We thank X. Ye and W. Wei for reagents; L. Wang, N. Ding and X. Yang for technical assistance; and Yuhang Zhang and G. Zhang for editing the manuscript. We thank the Laboratory Animal Resources Center, Tsinghua University for their support.
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X.T. conceived and supervised the project. M.M., Y. Ya., Zihan Zhang, Y. Yi., J.T., J.C., Y.G., Z.W., D.W., Y.L., Y.C. and H.W. conducted the experiments. Zhiyuan Zhang, L.L. and Y.P. contributed to the computational analysis. R.L., Q.H., Z.D., X.H., C.S., Y.Z, J.L., W.T., B.L., L.M. and G.C. provided key reagents or analysis and facility support. M.M. and X.T. wrote the manuscript with input from all other authors.
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Extended data
Extended Data Fig. 1 Different major isoforms of LRP8 have similar activities in promoting YF infection.
a, Schematic structure of LRP8 isoforms; b, In HEK293T cells, luciferase, LRP8 isoforms and its truncations were transiently overexpressed and the cells are infected by YFV 17D pseudoviruses. The ratios of infected cells were analyzed using FACS at 24hpi. c,d, Expression of the isoforms and its truncations on cell surface were determined with anti-LRP8 by FACS. e, f, Expression of luciferase, LDLR, LRP8 or TIM1 containing 3xHA tag overexpressed in HEK293T were analyzed by Western blot (e) or FACS (f). g. In HEK293T cells, luciferase, LRP8-ΔLBD and LRP8 were transiently overexpressed and the cells are infected by live YF17D with different MOI for 48 h. The infected cells were analyzed using Western blots. Statistical analysis was performed using one-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons test(b). The statistics shown are Mean ± SEM. NS: not significant. The replicate in the figure indicates biological replicates, the experiments were repeated at least twice.
Extended Data Fig. 2 LRP8 promotes YF17D infection in K562 cells.
a, b. Ectopic LRP8 (a) and luciferase (b) expression on K562 cell surface were determined by FACS with HA antibody staining. c. In K562 cells, HA-tagged luciferase and LRP8 were stably overexpressed and the cells are infected by YFV 17D pseudoviruses and the infection rates were determined by FACS at 24 hpi. d. In K562 cells, HA-tagged luciferase and LRP8 were stably overexpressed and the cells are infected by live YFV 17D and viral RNA level in cells were determined by RT-qPCR at 48 hpi. e. Infectious viral particles from d were determined by plaque assay. Statistical analysis was performed using two-way analysis of variance (ANOVA) with Dunnett’s multiple comparisons test(c-e). The statistics shown are Mean ± SEM. NS: not significant. The replicate in the figure indicates biological replicates, the experiments were repeated at least twice.
Extended Data Fig. 3 LRP8 promotes the infection by live viruses of YF17D, BJ01 and BJ01-GFP pseudovirus.
a. Supernatant of HEK293T expressing Luciferase and LRP8 infected with YF17D (0.05 MOI) and BJ01(0.05 MOI). At 48hpi, the viral titers were determined by plaque assay in BHK21 cells. b. In HEK293T cells, luciferase, LRP8-ΔLBD and LRP8 were transiently overexpressed and the cells are infected by live BJ01 with different MOI for 48 h. The infected cells were analyzed using Western blots. c. In HEK293T cells, luciferase and LRP8 were transiently overexpressed and the cells are infected by YFV BJ01 pseudoviruses. The ratios of infected cells were analyzed using FACS at 24 hpi. Statistical analysis was performed using two-way analysis of variance (ANOVA) with Uncorrected Fisher’s LSD test (a), Dunnett’s multiple comparisons test(c). The statistics shown are Mean ± SEM. NS: not significant. The replicate in the figure indicates biological replicates, the experiments were repeated at least twice.
Extended Data Fig. 4 Deficiency of LRP8 decreases live Asibi and YF17D infection.
a. U-87 MG cells clones expressing Cas9/sgLRP8 or Cas9 only were tested by FACS after staining with anti-LRP8 antibody. b, LRP8 KO or control U-87 MG cells were infected with the clinical Asibi (0.2 moi), viral RNA was quantified by RT–qPCR. c-e, In two clones LRP8-KO or control U-87 MG cells, LRP8-ΔLBD-mCherry and LRP8-mcherry plasmids were electroporated and 48 h later the cells are infected with YF17D. At 48 hpi, the cells were stained for YFV E protein and quantitated by fluorescence microscope. ΔLBD-mCherry and LRP8-mcherry expressions were determined by FACS. f. U-87 MG-sgLRP8 and control U-87 MG cells expressing only Cas9 were preincubated with 10 μg/ml anti-LRP8 (sc-1A1) or IgG before being infected by YF17D. At 48 hpi, the infections were analyzed by RT-qPCR. Statistical analysis was performed using two-way analysis of variance (ANOVA) with Turkey’s multiple comparisons test (b, c, f). The statistics shown are Mean ± SEM. NS: not significant. The replicate in the figure indicates biological replicates, the experiments were repeated at least twice.
Extended Data Fig. 5 LRP8 LBD binds 17D viral particles.
The binding of YFV 17D viral particles to purifed LBD domains of LRP8 or LDLR was measured by BLI assay.
Extended Data Fig. 6 Expression of different members of the LDLR family and LRP8 orthologs from different species.
a, In HEK293T cells, plasmids expressing different LDLR family members were transfected into HEK293T for 48 h and the target proteins containing 3X HA tag were analyzed by Western blot. The corresponding molecular weight of each protein were marked. b, In HEK293T cells, plasmids expressing different LRP8 orthologs were transfected into HEK293T for 48 h and the target proteins containing 3X HA tag were analyzed by Western blot.
Extended Data Fig. 7 The amino acid sequence alignment of LRP8 ligand binding domain (LBD).
The amino acid sequences of LRP8 ligand binding domains of human, mouse, mosquito, A. mississippiensis, bat, chicken, bird, dog, horse, green monkey were downloaded from NCBI and aligned using Geneious software.
Extended Data Fig. 8 LRP8 is expressed in human liver.
LRP8 expression were determined in human liver sample by immunohistochemistry staining with a LRP8 antibody (Thermo 3H2 clone).
Extended Data Fig. 9 Soluble LRP8-Fc blocks BJ01 and YF17D infection.
After preincubation of soluble LRP8 (200 μg/ml) with 0.5 MOI of BJ01 and YF17D for 1 h, the protein-virus mixture were added into PHHs for 48 h. The cellular viral RNA was quantified by RT-qPCR (three biological repeats).
Extended Data Fig. 10 Validation of the expression of LDLR and LRP8 delivered by AAV in mouse liver.
a, hLDLR and hLRP8 in mouse liver were determined at 3 weeks post AAV infection by Western blot; b, In the experiment of Fig. 4h, the livers of a mock mouse and two mice each from the LDLR and LRP8 group that paralyzed at day 7 (LRP8 group) or day 10 (LDLR group) were processed for immunohistochemistry imaging with DAB staining of YFV E protein with an anti-E antibody.
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Mei, M., Yang, Y., Zhang, Z. et al. LRP8 is a functional receptor for yellow fever virus. Nat Microbiol (2026). https://doi.org/10.1038/s41564-026-02278-7
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DOI: https://doi.org/10.1038/s41564-026-02278-7
