Data from: CARD8 inflammasome activation during HIV-1 cell-to-cell transmission
Data files
Jun 12, 2025 version files 16.95 MB
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20250611_uncropped_westerns.pdf
16.91 MB
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eLife_Kulsuptrakul_Dryad.xlsx
30.41 KB
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ICE_Primers.xlsx
9.53 KB
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README.md
2.05 KB
Abstract
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.
Dataset DOI: 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.