DOI: 10.5061/dryad.qbzkh18vn
title: Data from: CARD8 inflammasome activation during HIV-1 cell-to-cell transmission

description: 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
 affects CARD8 activation. We identified mutant HIV-1 proteases that
 differentially cleave and activate CARD8 compared to wildtype HIV-1, thus
 indicating that natural variation in HIV protease affects not only the
 cleavage of the viral Gag-Pol polyprotein but also likely impacts innate
 sensing and inflammation.
description: # 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. Dashed red box indicates lanes used for the figure.
 #### File: ICE\_Primers.xlsx **Description:** list of primers used for
 knockout genotyping for Synthego ICE analysis ##### Variables * N/A ##
 Access information Other publicly accessible locations of the data: * new
 sequencing files for the protease inhibitor resistant strains of HIV-1 can
 be found on GenBank under accession numbers: BankIt2967949 PI-R1 PV761644,
 BankIt2967949 PI-R2 PV761645, BankIt2967949 PI-R5 PV761646, BankIt2967949
 PI-R9 PV761647, BankIt2967949 PI-R10 PV761648, BankIt2967949 PI-R12
 PV761649, BankIt2967949 PI-R13 PV761650 Data was derived from the
 following sources: * Reported mutations in Table S1 from the protease
 inhibitor resistant strains of HIV were derived from the NIH AIDS
 repository from Varghese et al 2013. However, we also did whole plasmid
 sequencing on these constructions and uncovered some additional unreported
 mutations in these constructions. These new sequencing files can be found
 under the GenBank accession numbers listed above.