{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T03:59:57Z","timestamp":1788235197850,"version":"build-2803163510"},"reference-count":29,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,6,12]],"date-time":"2022-06-12T00:00:00Z","timestamp":1654992000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01AI153348"],"award-info":[{"award-number":["R01AI153348"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01AI150931"],"award-info":[{"award-number":["R01AI150931"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000060","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R21AI147152"],"award-info":[{"award-number":["R21AI147152"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Human cytomegalovirus (CMV) is a ubiquitous pathogen that latently resides in hematopoietic cells. Latently infected individuals with dysfunctional immune systems often experience CMV reactivation, which can cause devastating disease and mortality. While factors dictating the balance between latency and reactivation are not completely understood, CMV US28 is required for maintaining latent infection, and viral mutants that alter US28 function result in a lytic-like, rather than latent, infection in hematopoietic cells. In turn, viral lytic factors alter the host cell, making it challenging to characterize the US28-specific changes in the cellular milieu. To circumvent this, we generated a temperature-sensitive TB40\/E recombinant virus, TB40\/EgfpC510G (tsC510G), into which we engineered an amino acid change at position 510 (C510G) of IE2, as previously described in the CMV Towne strain. Using tsC510G, we then deleted the US28 ORF, termed tsC510G-US28\u0394. Consistent with previous findings, tsC510G-US28\u0394 fails to undergo latency in Kasumi-3 cells at the permissive temperature. However, parallel cultures maintained at the non-permissive temperature showed a significant reduction in infectious center frequency, as measured by limiting dilution assay. Thus, we generated a new US28 mutant virus for use as a tool to study US28-specific changes in latently infected hematopoietic cells in the absence of induced lytic replication.<\/jats:p>","DOI":"10.3390\/v14061280","type":"journal-article","created":{"date-parts":[[2022,6,13]],"date-time":"2022-06-13T06:31:59Z","timestamp":1655101919000},"page":"1280","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Design of a US28 ORF Deletion Virus in a Temperature-Sensitive Cytomegalovirus Strain Fails to Promote Lytic Replication in Hematopoietic Cells"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0919-2961","authenticated-orcid":false,"given":"Benjamin A.","family":"Krishna","sequence":"first","affiliation":[{"name":"Genomic Medicine, Infection Biology Program, Global Center for Pathogen & Human Health Research, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7712-6259","authenticated-orcid":false,"given":"Amanda B.","family":"Wass","sequence":"additional","affiliation":[{"name":"Genomic Medicine, Infection Biology Program, Global Center for Pathogen & Human Health Research, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eain A.","family":"Murphy","sequence":"additional","affiliation":[{"name":"Microbiology and Immunology, Upstate Medical University\u2014State University of New York (SUNY), Syracuse, NY 13210, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4943-3774","authenticated-orcid":false,"given":"Christine M.","family":"O\u2019Connor","sequence":"additional","affiliation":[{"name":"Genomic Medicine, Infection Biology Program, Global Center for Pathogen & Human Health Research, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1182\/blood-2002-03-0993","article-title":"Late cytomegalovirus disease and mortality in recipients of allogeneic hematopoietic stem cell transplants: Importance of viral load and T-cell immunity","volume":"101","author":"Boeckh","year":"2003","journal-title":"Blood"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2003","DOI":"10.1182\/blood-2003-10-3616","article-title":"The impact of cytomegalovirus serostatus of donor and recipient before hematopoietic stem cell transplantation in the era of antiviral prophylaxis and preemptive therapy","volume":"103","author":"Boeckh","year":"2004","journal-title":"Blood"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1111\/j.1399-3062.2010.00504.x","article-title":"Pre-transplant cytomegalovirus (CMV) serostatus remains the most important determinant of CMV reactivation after allogeneic hematopoietic stem cell transplantation in the era of surveillance and preemptive therapy","volume":"12","author":"George","year":"2010","journal-title":"Transpl. 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