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…

Disclosures

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

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Cite This Article
Oral Combinational Antiretroviral Therapy to Treat HIV-1 Infection in a Humanized Mouse Model. J. Vis. Exp. (Pending Publication), e22229, doi: (2024).

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