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Updated: Jun 20, 2026

Retroviral Transduction of T-cell Receptors in Mouse T-cells
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Efficient Retroviral Transduction and Competitive Homing for Investigating GPCR-Mediated T-Cell Localization in

Aiman Ayesha1, Borja Ocon2, Yuhan Bi2

  • 1Department of Pathology, Stanford University School of Medicine; Palo Alto Veterans Institute for Research; Veterans Affairs Palo Alto Health Care System; aiman@stanford.edu.

Journal of Visualized Experiments : Jove
|April 14, 2025
PubMed

Understanding how G-protein coupled receptor (GPCR) expression affects cell positioning within diverse tissue microenvironments is essential for elucidating immune cell trafficking mechanisms. We present a competitive homing assay designed to study GPCR-mediated T-cell localization to organs expressing their cognate chemoattractant ligands, applicable for both short-term and long-term studies. The approach involves an improved protocol for recombinant murine stem cell virus (MSCV) transduction of T cells to express the GPCR of interest or a control construct, followed by competitive homing in recipient mice. Cell distribution across different organs is analyzed using flow cytometry and/or confocal microscopy. In short-term experiments (10-12 h), confocal microscopy revealed distinct cell localization patterns, including to alveoli, bronchi submucosa, venous sites, and interstitium in the lung, as well as the epithelium lining the trachea, stomach, and uterine horn. In long-term studies (1-7 weeks), flow cytometry provided insights into preferential cell accumulation, revealing dynamic changes and potential maturation or repositioning within tissues over time. This competitive homing assay is a robust tool for studying GPCR-mediated cell positioning, offering valuable insights into tissue-specific distribution and potential applications in immunology and therapeutic research.

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