title: Longitudinal imaging of HIV-1 spread in humanized mice with parallel 3D immunofluorescence and electron tomography.
abstract: Dissemination of HIV-1 throughout lymphoid tissues leads to systemic virus spread following infection. We combined tissue clearing, 3D-immunofluorescence, and electron tomography (ET) to longitudinally assess early HIV-1 spread in lymphoid tissues in humanized mice. Immunofluorescence revealed peak infection density in gut at 10-12 days post-infection when blood viral loads were low. Human CD4+ T-cells and HIV-1-infected cells localized predominantly to crypts and the lower third of intestinal villi. Free virions and infected cells were not readily detectable by ET at 5-days post-infection, whereas HIV-1-infected cells surrounded by pools of free virions were present in ~10% of intestinal crypts by 10-12 days. ET of spleen revealed thousands of virions released by individual cells and discreet cytoplasmic densities near sites of prolific virus production. These studies highlight the importance of multiscale imaging of HIV-1-infected tissues and are adaptable to other animal models and human patient samples.
authors.affiliation: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States.
authors.forename: Collin
authors.initials: C
authors.lastname: Kieffer
authors.affiliation: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Ladinsky
authors.affiliation: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States.
authors.forename: Allen
authors.initials: A
authors.lastname: Ninh
authors.affiliation: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States.
authors.forename: Rachel P
authors.initials: RP
authors.lastname: Galimidi
authors.affiliation: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States.
authors.forename: Pamela J
authors.identifier: 0000-0002-2277-3990
authors.initials: PJ
authors.lastname: Bjorkman
mesh_terms: D000825:Animal Structures / Q000821:virology; D000818:Animals; D001826:Body Fluids / Q000821:virology; D055032:Electron Microscope Tomography; D005455:Fluorescent Antibody Technique; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000254:growth & development*; D021621:Imaging, Three-Dimensional; D008137:Longitudinal Studies; D016513:Mice, SCID; D013997:Time Factors
keywords: 3D immunofluorescence; CLARITY; HIV-1; electron tomography; humanized mouse; infectious disease; microbiology; mouse; tissue clearing