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Related Experiment Video

Updated: Jan 6, 2026

Humanized NOG Mice for Intravaginal HIV Exposure and Treatment of HIV Infection
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Humanizing Mice for Human Immunodeficiency Virus Infection Studies.

Chandra Nath Roy1, Christopher Kline1, Ashley Gnanasekaran1

  • 1Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania.

Current Protocols
|November 25, 2025
PubMed
Summary

This study details protocols for creating humanized mice to model human immunodeficiency virus type 1 (HIV-1) infection. These methods enable robust HIV-1 research, including viral challenge and precise RNA detection in vivo.

Keywords:
HIVRT‐qPCRflow cytometryhumanized miceinfectiontransmission

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Chronic, Acute, and Reactivated HIV Infection in Humanized Immunodeficient Mouse Models
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Area of Science:

  • Immunology
  • Virology
  • Transplantation Biology

Background:

  • Human immunodeficiency virus type 1 (HIV-1) infects an estimated 40 million people globally, with no effective cure.
  • Humanized mice are critical preclinical models for studying HIV-1 pathogenesis, transmission, and therapeutic interventions.
  • Existing models require refined protocols for efficient human immune cell engraftment and reproducible HIV-1 infection.

Purpose of the Study:

  • To provide comprehensive protocols for generating humanized immunodeficient mouse models.
  • To establish standardized methods for HIV-1 production, challenge, and quantification in these models.
  • To facilitate reproducible research on HIV-1 infection dynamics and treatment efficacy.

Main Methods:

  • Isolation of CD34+ hematopoietic stem cells from human cord blood.
  • Engraftment of stem cells into immunodeficient mice (e.g., NSG, MISTRG6 strains).
  • In vitro production and titration of HIV-1, followed by systemic or mucosal challenge.
  • Flow cytometry for immune cell reconstitution assessment and RT-qPCR for HIV-1 RNA detection.

Main Results:

  • Successful human immune cell reconstitution in immunodeficient mice.
  • Establishment of protocols for producing and titering infectious HIV-1.
  • Demonstration of sensitive HIV-1 RNA detection in mouse plasma using RT-qPCR with an internal control.
  • Validation of both systemic and mucosal challenge models.

Conclusions:

  • The provided protocols enable the generation of reliable humanized mouse models for HIV-1 research.
  • These methods support the study of HIV-1 infection, pathogenesis, and the evaluation of potential therapeutics.
  • The developed techniques facilitate longitudinal monitoring of viral load in preclinical HIV-1 studies.