| Autor: |
Andersen AHF; Department of Clinical Medicine, Aarhus University; Department of Infectious Diseases, Aarhus University Hospital; ahfa@clin.au.dk., Nielsen SSF; Department of Clinical Medicine, Aarhus University; Department of Infectious Diseases, Aarhus University Hospital., Olesen R; Department of Clinical Medicine, Aarhus University; Department of Infectious Diseases, Aarhus University Hospital., Mack K; Department of Clinical Medicine, Aarhus University., Dagnæs-Hansen F; Department of Biomedicine, Aarhus University., Uldbjerg N; Department of Clinical Medicine, Aarhus University; Department of Gynecology and Obstetrics, Aarhus University Hospital., Østergaard L; Department of Clinical Medicine, Aarhus University; Department of Infectious Diseases, Aarhus University Hospital., Søgaard OS; Department of Clinical Medicine, Aarhus University; Department of Infectious Diseases, Aarhus University Hospital., Denton PW; Department of Clinical Medicine, Aarhus University; Department of Infectious Diseases, Aarhus University Hospital; Department of Biology, University of Nebraska at Omaha., Tolstrup M; Department of Clinical Medicine, Aarhus University; Department of Infectious Diseases, Aarhus University Hospital. |
| Abstrakt: |
Humanized mice provide a sophisticated platform to study human immunodeficiency virus (HIV) virology and to test antiviral drugs. This protocol describes the establishment of a human immune system in adult NOG mice. Here, we explain all the practical steps from isolation of umbilical cord blood derived human CD34+ cells and their subsequent intravenous transplantation into the mice, to the manipulation of the model through HIV infection, combination antiretroviral therapy (cART), and blood sampling. Approximately 75,000 hCD34+ cells are injected intravenously into the mice and the level of human chimerism, also known as humanization, in the peripheral blood is estimated longitudinally for months by flow cytometry. A total of 75,000 hCD34+ cells yields 20%-50% human CD45+ cells in the peripheral blood. The mice are susceptible to intravaginal infection with HIV and blood can be sampled once weekly for analysis, and twice monthly for extended periods. This protocol describes an assay for quantification of plasma viral load using droplet digital PCR (ddPCR). We show how the mice can be effectively treated with a standard-of-care cART regimen in the diet. The delivery of cART in the form of regular mouse chow is a significant refinement of the experimental model. This model can be used for preclinical analysis of both systemic and topical pre-exposure prophylaxis compounds as well as for testing of novel treatments and HIV cure strategies. |
