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Merck: Investigating the Active HIV Reservoir

Writer's picture: Ray SullivanRay Sullivan

Updated: Feb 2

Guoxin Wu’s group at Merck Research Labs in Rahway, NJ (designated as a “Milestones in Microbiology” site by ASM) developed an immunocytochemistry method called CA-ICC to detect and quantify intracellular HIV capsid protein (CA) in bulk cell preparations, which was shown to be specific and sensitive.  Fixed cells are deposited on microscopy slides using a cytospin technique and then stained with two anti-CA monoclonal antibodies using an automated stainer.  The slides are then digitized and analyzed using computational image analysis to quantify the number of CA-positive cells.  CA-ICC detected 10 CA-positive cells per million, which was more sensitive than flow cytometry.  CA-ICC was used to measure the activity of a targeted activator of cell kill (TACK) compound, which selectively eliminated CA-expressing cells, and the results were comparable to other standard assays.  CA-ICC provides a simple, sensitive, and rapid alternative to existing assays for quantifying the translationally active HIV reservoir, with potential applications in preclinical and clinical research.


Sensitivity and specificity of CA-ICC. (A) MOLT IIIB cells stained with mIgG or (B) anti-CA antibodies. (C) Jurkat cells stained with anti-CA antibodies. (D) Uninfected and (E) in vitro infected human PBMCs stained with anti-CA antibodies. Representative raw images (left panels) and relative computational markup (right panels) are shown. Markup colors denote cells with strong (red), moderate (orange), and weak (yellow) chromogenic signal intensity. The top and bottom images for each panel represent 20× and 80× magnification, respectively.
Sensitivity and specificity of CA-ICC. (A) MOLT IIIB cells stained with mIgG or (B) anti-CA antibodies. (C) Jurkat cells stained with anti-CA antibodies. (D) Uninfected and (E) in vitro infected human PBMCs stained with anti-CA antibodies. Representative raw images (left panels) and relative computational markup (right panels) are shown. Markup colors denote cells with strong (red), moderate (orange), and weak (yellow) chromogenic signal intensity. The top and bottom images for each panel represent 20× and 80× magnification, respectively.

Keller SH, Deng H, Lim B. Regulation of the dynamic RNA Pol II elongation rate in Drosophila embryos. Cell Rep. 2023 Oct 31;42(10):113225. doi: 10.1016/j.celrep.2023.113225. Epub 2023 Oct 12. PMID: 37837623; PMCID: PMC10842316.  https://www.mdpi.com/1422-0067/26/2/682

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