Abstract
Human cytomegalovirus (HCMV) infection results in either productive or latent infection, the latter enabling life-long viral persistence. Monocytes support latent infection but become permissive to productive infection upon differentiation into macrophages. These differentiation-driven differences have been largely attributed to chromatin-mediated repression of the viral genome. Using metabolic labeling of newly synthesized RNA, we observe markedly lower viral transcription at early stages of infection in monocytes compared to macrophages. Unbiased comparison reveals that this difference is partly explained by inefficient viral entry in monocytes: fewer viruses enter, and correspondingly, fewer genomes reach the nucleus. Indeed, ectopic expression of known HCMV entry receptors in monocytes enhances viral entry and enables productive infection, demonstrating that these cells can support full lytic replication if entry is efficient. We further identify integrin β3 as a differentiation-induced surface protein playing an important role in HCMV entry into macrophages, partially accounting for the observed differences in entry efficiency. Finally, we show that cells receiving fewer viral genomes are the ones that establish latent infection and have the capacity to reactivate. Overall, our findings reveal that entry is a previously unrecognized factor contributing to latent infection in monocytes, adding a critical layer to the paradigm of HCMV latency.
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Data availability
All next-generation sequencing data files have been deposited in Gene Expression Omnibus under accession number GSE280650. Mass spectrometry data are available via ProteomeXchange with identifier PXD071694. Source data are provided with this paper.
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Acknowledgements
We thank Dr. Orly Laufman, Prof. Yossef Shaul and the members of the Stern-Ginossar lab for the critical reading of the manuscript. We thank Dr. Dor Simkin, Dr. Avner Leshem and Tatiana Smirnova for technical assistance. All optical imaging acquired at the de Picciotto Cancer Cell Observatory in memory of Wolfgang and Ruth Lesser of the Moross Integrated Cancer Center in the Department of Life Science Core Facilities, Weizmann Institute of Science. We would like to thank the Weizmann flow cytometry and microscopy unit for technical assistance. We’d like to thank David Morgenstern from the protein profile unit at the G-INCPM, the Weizmann Institute, for his assistance with proteomics analysis. This study was supported by a European Research Council consolidator grant (CoG-2019-864012) and an Israel Science Foundation grant to N.S.-G. (2507/23).
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Y.K., N.S.-G., and M.S. conceived and designed the project. Y.K., T.A., A.W., and M.S. performed the experiments. T.F. helped analyze the HCMV particle images. K.B prepared the RNA for the transfections experiments. Y.F. offered valuable guidance throughout the research. Y.K., A.N., N.S-G., and M.S. analyzed and interpreted the data. Y.K., N.S.-G., and M.S. wrote the manuscript with input from all the authors.
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Kitsberg, Y., Nachshon, A., Arazi, T. et al. Viral entry shapes HCMV latency establishment. Nat Commun (2025). https://doi.org/10.1038/s41467-025-68063-y
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DOI: https://doi.org/10.1038/s41467-025-68063-y
