Oral Combinational Antiretroviral Therapy to Treat HIV-1 Infection in a Humanized Mouse Model
Published: April 30, 2024
Abstract
Source: Mu, W., et al. Oral Combinational Antiretroviral Treatment in HIV-1 Infected Humanized Mice. J. Vis. Exp. (2022).
This video demonstrates an assay to evaluate the effectiveness of oral combined antiretroviral therapy (oral cART) in a human immunodeficiency virus-1 (HIV-1)-infected humanized mouse model. A combination of drugs targeting HIV-1, including nucleoside reverse transcriptase inhibitors and integrase strand transfer inhibitors, is administered along with a nutritional supplement and antibiotics. Following the consumption of this mixture by the humanized mice infected with HIV-1, the effective suppression of viral replication is assessed through periodic examinations of the mice.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Humanized mice infected with HIV-1 NOTE: Humanized mice were constructed as previously described. The protocol is briefly described below. Purify CD34+ hematopoietic progenitor cells from the human fetal liver via anti-CD34 microbeads according to the manuf…
Divulgations
The authors have nothing to disclose.
Materials
| 60 mm petri dish | Thermo Scientific Nunc | 150288 | For aliquoting ART food |
| APC anti-human CD8 Antibody | Biolegend | 344722 | For flow cytometry |
| BD LSRFortessa | BD biosciences | For flow data collection | |
| CD34 microbeads | Miltenyi Biotec | 130-046-702 | For NSG-BLT mice generation |
| Centrifuge tubes | Falcon | 14-432-22 | For dissolving ART |
| DietGel Boost | ClearH2O | 72-04-5022 | For making ART food |
| Elvitegravir | Gilead | Gifted from Gilead | |
| Emtricitabine | Gilead | Gifted from Gilead | |
| FITC anti-human CD3 Antibody | Biolegend | 317306 | For flow cytometry |
| Flowjo software | FlowJo | For flow cytometry data analysis | |
| HIV-1 forward primer: 5'-CAATGGCAGCAATTTCACCA-3'; | IDT | Customized | For viral load RT-PCR |
| HIV-1 probe: 5'-[6-FAM]CCCACCAACAGGCGGCCT TAACTG [Tamra-Q]-3'; |
IDT | Customized | For viral load RT-PCR |
| HIV-1 reverse primer: 5'-GAATGCCAAATTCCTGCTTGA-3'; | IDT | Customized | For viral load RT-PCR |
| Human fetal tissue | Advanced Bioscience Resources, Inc | ||
| Mice, strain NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ | The Jackson Laboratory | 5557 | For constructing the humanized mice |
| Pacific Blue anti-human CD45 | Biolegend | 304022 | For flow cytometry |
| PerCP anti-human CD4 Antibody | Biolegend | 300528 | For flow cytometry |
| QIAamp Viral RNA Kits | Qiagen | 52904 | For measuring viral load |
| Raltegravir | Merck | Gifted from Merck | |
| Sterile cell scrapers | Thermo Scientific | 179693 | For aliquoting ART food |
| TaqMan RNA-To-Ct 1-Step Kit | Applied Biosystems | 4392653 | For plasma viral load detection |
| Tenofovir disoproxil fumarate | Gilead | Gifted from Gilead | |
| Trimethoprim-Sulfamethoxazole | Pharmaceutical Associates | NDC 0121-0854-16 | For keeping ART food sterile. Each 5mL teaspoon contains 200 mg Sulfamethoxazole, USP 40 mg Trimethoprim, USP NMT 0.5% Alcohol |
Tags
Citer Cet Article
Oral Combinational Antiretroviral Therapy to Treat HIV-1 Infection in a Humanized Mouse Model. J. Vis. Exp. (Pending Publication), e22229, doi: (2024).