{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T20:30:06Z","timestamp":1785529806098,"version":"3.56.0"},"reference-count":20,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T00:00:00Z","timestamp":1764028800000},"content-version":"vor","delay-in-days":24,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2025,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>An estimated 40 million people are infected with human immunodeficiency virus type 1 (HIV\u20101) and there is no effective cure. Humanized mice are a useful model to study transmission, pathogenesis, prevention, and treatment of HIV\u20101, which only infects human cells. Here we provide detailed protocols to humanize immunodeficient mice, e.g., NSG or MISTRG6 strains, using human hematopoietic stem cells isolated from cord blood and a flow cytometry panel of antibodies to assess human immune cell reconstitution. Furthermore, we describe how to produce, concentrate, and titer HIV\u20101 in vitro, which can be used to challenge humanized mice either systemically or mucosally. Finally, we adapted a sensitive quantitative reverse transcription\u2013polymerase chain reaction (RT\u2010qPCR) assay with internal control to detect HIV\u20101 RNA in longitudinal plasma samples to measure viremia over time in the mice. \u00a9 2025 Wiley Periodicals LLC.<\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 1<\/jats:bold>\n                    : Isolation of CD34+ cells from human cord blood\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 2<\/jats:bold>\n                    : Injection of human CD34+ cells in immunodeficient mice\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 3<\/jats:bold>\n                    : Flow cytometry of peripheral blood mononuclear cells from mouse blood\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 4<\/jats:bold>\n                    : Production and titration of HIV\u20101\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 5<\/jats:bold>\n                    : Systemic HIV\u20101 challenge of humanized mice\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Alternate Protocol<\/jats:bold>\n                    : Mucosal HIV\u20101 challenge of humanized mice\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Basic Protocol 6<\/jats:bold>\n                    : Single\u2010copy HIV\u20101 RNA detection in mouse plasma\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 1<\/jats:bold>\n                    : Retrovirus internal control for plasma virus isolation and RT\u2010qPCR\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Support Protocol 2<\/jats:bold>\n                    : Production of HIV\u20101 standard for RT\u2010qPCR\n                  <\/jats:p>","DOI":"10.1002\/cpz1.70266","type":"journal-article","created":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T10:22:58Z","timestamp":1764066178000},"source":"Crossref","is-referenced-by-count":1,"title":["Humanizing Mice for Human Immunodeficiency Virus Infection Studies"],"prefix":"10.1002","volume":"5","author":[{"given":"Chandra Nath","family":"Roy","sequence":"first","affiliation":[{"name":"Department of Microbiology and Molecular Genetics University of Pittsburgh School of Medicine  Pittsburgh Pennsylvania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher","family":"Kline","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics University of Pittsburgh School of Medicine  Pittsburgh Pennsylvania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ashley","family":"Gnanasekaran","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics University of Pittsburgh School of Medicine  Pittsburgh Pennsylvania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8610-2766","authenticated-orcid":false,"given":"Zandrea","family":"Ambrose","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Molecular Genetics University of Pittsburgh School of Medicine  Pittsburgh Pennsylvania"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,11,25]]},"reference":[{"key":"e_1_2_15_2_1","doi-asserted-by":"publisher","DOI":"10.5772\/33484"},{"key":"e_1_2_15_3_1","doi-asserted-by":"publisher","DOI":"10.2217\/17469600.2.4.363"},{"key":"e_1_2_15_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.clim.2009.12.008"},{"key":"e_1_2_15_5_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.72.9.6988\u20106996.1998"},{"key":"e_1_2_15_6_1","doi-asserted-by":"publisher","DOI":"10.1136\/jitc\u20102023\u2010006921"},{"key":"e_1_2_15_7_1","doi-asserted-by":"publisher","DOI":"10.1002\/9780471729259.mca02as00"},{"key":"e_1_2_15_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41684\u2010023\u201001196\u2010z"},{"issue":"3","key":"e_1_2_15_9_1","first-page":"135","article-title":"Humanized mouse models of HIV infection","volume":"13","author":"Denton P. 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