A computer-based, image-analysis method to quantify HIV-1 infection in a single-cycle infectious center assay

Area of Science:

• Virology
• Cell Biology
• Biotechnology

Background:

• Accurate quantitation of human immunodeficiency virus (HIV) infectivity is essential for advancing molecular virology research.
• Measuring the efficacy of antiviral drugs and neutralizing antibodies requires precise HIV infectivity assays.

Purpose of the Study:

• To develop and validate a novel method for rapid and precise quantitation of HIV infectivity.
• To utilize an indicator cell line and image analysis software for quantifying HIV infection levels.

Main Methods:

• Employing the P4R5 indicator cell line, which expresses HIV receptors and beta-galactosidase.
• Infecting cells with CXCR4- or CCR5-using HIV strains, followed by X-gal staining to visualize infected cells.
• Utilizing a high-resolution CCD camera and macro lens for image capture, and custom software for automated counting of blue-stained foci.

Main Results:

• The developed assay enables rapid quantitation of HIV-infected cells across a broad range of viral inocula.
• The method demonstrates reproducibility and accuracy in assessing infectivity.
• The assay was successfully applied to evaluate viral mutants, antiviral drugs, and neutralizing antibodies.

Conclusions:

• The described image-analysis-based method provides a rapid, reproducible, and accurate means for quantifying HIV infectivity.
• This cost-effective assay is suitable for various applications in HIV research and drug screening.