CARD8 inflammasome activation during HIV-1 cell-to-cell transmission

Our previous work demonstrated that CARD8 detects HIV-1 infection by sensing the enzymatic activity of the HIV protease, resulting in CARD8-dependent inflammasome activation (Kulsuptrakul et al., 2023). CARD8 harbors a motif in its N-terminus that functions as a HIV protease substrate mimic, permitting innate immune recognition of HIV-1 protease activity, which when cleaved by HIV protease triggers CARD8 inflammasome activation. Here, we sought to understand CARD8 responses in the context of HIV-1 cell-to-cell transmission via a viral synapse. We observed that cell-to-cell transmission of HIV-1 between infected T cells and primary human monocyte-derived macrophages induces CARD8 inflammasome activation in a manner that is dependent on viral protease activity and largely independent of the NLRP3 inflammasome. Additionally, to further evaluate the viral determinants of CARD8 sensing, we tested a panel of HIV protease inhibitor resistant clones to establish how variation in HIV protease af..., , # CARD8 inflammasome activation during HIV-1 cell-to-cell transmission

Dataset DOI: [10.5061/dryad.qbzkh18vn](10.5061/dryad.qbzkh18vn)

## Description of the data and file structure

This data was collected to assay CARD8 inflammasome activation during HIV-1 cell-to-cell transmission. We assessed CARD8 inflammasome activation in both cancer and primary myeloid cells.

### Files and variables

#### File: eLife\_Kulsuptrakul\_Dryad.xlsx

**Description:** The excel file includes tabs corresponding to each figure of the paper in the closest format to how the data was graphed and presented in the manuscript.

##### Variables

* Knockout cells: CARD8 KO, AAVS1 KO, NLRP3 KO or wildtype cells
* Drug inhibitor treatments: DMSO, RT inhibitor (NVP), lopinavir (LPV), VX765, MCC950
* Drug treatment: VbP or nigericin treatment

#### File: 20250507\_uncropped\_westernblots.pdf

**Description:** Uncropped western blots multichannel images shown are a merge of chemiluminescent and colorimetric images. ...,