Flow Cytometry-Based Quantification of HIV-Induced T Cell Chemotactic Response

Area of Science:

• Immunology
• Virology
• Cell Biology

Background:

• Human immunodeficiency virus (HIV) infects CD4 T cells by binding to CD4 and chemokine co-receptors (CXCR4 or CCR5).
• Viral binding initiates signaling pathways involving LIMK/cofilin and WAVE2/Arp2/3, promoting actin dynamics.
• Actin dynamics are crucial for viral nuclear migration and establishing latent infection in resting CD4 T cells.

Purpose of the Study:

• To describe methods for quantifying HIV-induced actin polymerization.
• To detail methods for measuring cofilin phosphorylation in human T cells.
• To provide a flow cytometry-based approach for analyzing HIV's impact on T cell actin dynamics.

Main Methods:

• Utilizing flow cytometry for cellular analysis.
• Quantifying actin polymerization in response to HIV infection.
• Measuring the phosphorylation levels of cofilin, a key actin-regulatory protein.

Main Results:

• The study outlines a flow cytometry protocol to measure HIV-induced changes in actin dynamics.
• Methods are presented for quantifying cofilin phosphorylation, a downstream event of viral entry.
• The described techniques allow for the assessment of molecular events supporting HIV latency.

Conclusions:

• Flow cytometry provides a robust method for quantifying HIV-induced actin polymerization and cofilin phosphorylation.
• Understanding these actin dynamics is key to deciphering mechanisms of HIV nuclear migration and latency.
• This work offers valuable tools for researchers studying HIV-1 infection and T cell responses.