, NALP3 forms an IL-1beta-processing inflammasome with increased activity in Muckle-Wells autoinflammatory disorder, Immunity, vol.20, pp.319-325, 2004.
, The TRIM family protein KAP1 inhibits HIV-1 integration, Cell Host Microbe, vol.9, pp.484-495, 2011.
, p21-mediated RNR2 repression restricts HIV-1 replication in macrophages by inhibiting dNTP biosynthesis pathway, Proc. Natl. Acad. Sci. USA, vol.110, pp.3997-4006, 2013.
URL : https://hal.archives-ouvertes.fr/pasteur-01420537
, The NLRP3 Inflammasome Is Upregulated in HIV-Infected Antiretroviral Therapy-Treated Individuals with Defective Immune Recovery, Mucosal SIV Infection of Rhesus Monkeys. Cell, vol.9, pp.656-667, 2016.
, Moesin is required for HIV-1-induced CD4-CXCR4 interaction, F-actin redistribution, membrane fusion and viral infection in lymphocytes, J. Cell Sci, vol.122, pp.103-113, 2009.
, Caspase-11 and caspase-1 differentially modulate actin polymerization via RhoA and Slingshot proteins to promote bacterial clearance, PLoS Pathog. 7, e1002347. Caution, vol.5, p.18479, 2011.
, Response of host inflammasomes to viral infection, Trends Microbiol, vol.23, pp.55-63, 2015.
,
, HIV-1 Tat Primes and Activates Microglial NLRP3 Inflammasome-Mediated Neuroinflammation, J. Neurosci, vol.37, pp.3599-3609
, T cell receptor activation leads to two distinct phases of Pyk2 activation and actin cytoskeletal rearrangement in human T cells, Mol. Immunol, vol.47, pp.1665-1674, 2010.
, The engagement of activating FcgammaRs inhibits primate lentivirus replication in human macrophages, J. Immunol, vol.177, pp.6291-6300, 2006.
URL : https://hal.archives-ouvertes.fr/pasteur-00167539
, The G140S mutation in HIV integrases from raltegravir-resistant patients rescues catalytic defect due to the resistance Q148H mutation, Nucleic Acids Res, vol.37, pp.1193-1201, 2009.
, Changes in soluble factor-mediated CD8+ cell-derived antiviral activity in cynomolgus macaques infected with simian immunodeficiency virus SIVmac251: relationship to biological markers of progression, J. Virol, vol.80, pp.236-245, 2006.
, Abortive HIV infection mediates CD4 T cell depletion and inflammation in human lymphoid tissue, Cell, vol.143, pp.789-801, 2010.
,
, Inhibition of PYK2-induced actin cytoskeleton reorganization, PYK2 autophosphorylation and focal adhesion targeting by FAK, J. Cell Sci, vol.114, pp.2977-2987, 2001.
, Nucleotides released by apoptotic cells act as a find-me signal to promote phagocytic clearance, Nature, vol.461, pp.282-286, 2009.
, The NALP3 inflammasome is involved in the innate immune response to amyloid-beta, Nat. Immunol, vol.9, pp.857-865, 2008.
, HIV Promotes NLRP3 Inflammasome Complex Activation in Murine HIV-Associated Nephropathy, vol.186, pp.347-358, 2016.
, The restriction factors of human immunodeficiency virus, J. Biol. Chem, vol.287, pp.40875-40883, 2012.
, Purinergic receptors are required for HIV-1 infection of primary human macrophages, J. Immunol, vol.188, pp.4488-4495, 2012.
, 3,4-methylenedioxy-b-nitrostyrene inhibits NLRP3 inflammasome activation by blocking assembly of the inflammasome, J. Biol. Chem, vol.289, pp.1142-1150, 2014.
, The inflammasome adaptor ASC regulates the function of adaptive immune cells by controlling Dock2-mediated Rac activation and actin polymerization, Nat. Immunol, vol.12, pp.1010-1016, 2011.
, Filamin-A regulates actin-dependent clustering of HIV receptors, Nat. Cell Biol, vol.9, pp.838-846, 2007.
, Trichothecene mycotoxins activate NLRP3 inflammasome through a P2X7 receptor and Src tyrosine kinase dependent pathway, Hum. Immunol, vol.75, pp.134-140, 2014.
, Src homology 3 binding sites in the P2Y2 nucleotide receptor interact with Src and regulate activities of Src, proline-rich tyrosine kinase 2, and growth factor receptors, J. Biol. Chem, vol.279, pp.8212-8218, 2004.
, HIV-1 Viral Protein R Activates NLRP3 Inflammasome in Microglia: implications for HIV-1 Associated Neuroinflammation, J. Neuroimmune Pharmacol, vol.12, pp.233-248, 2017.
, Visualization of the intracellular behavior of HIV in living cells, J. Cell Biol, vol.159, pp.441-452, 2002.
, IFI16 DNA sensor is required for death of lymphoid CD4 T cells abortively infected with HIV, Science, vol.343, pp.428-432, 2014.
, Pyk2 regulates multiple signaling events crucial for macrophage morphology and migration, Proc. Natl. Acad. Sci. USA, vol.100, pp.10740-10745, 2003.
, NF-kappaB and p53 are the dominant apoptosis-inducing transcription factors elicited by the HIV-1 envelope, J. Exp. Med, vol.199, pp.629-640, 2004.
, Essential role of p53 phosphorylation by p38 MAPK in apoptosis induction by the HIV-1 envelope, J. Exp. Med, vol.201, pp.279-289, 2005.
, A 3'UTR SNP in NLRP3 gene is associated with susceptibility to HIV-1 infection, J. Acquir. Immune Defic. Syndr, vol.54, pp.236-240, 2010.
, Cell Reports, vol.28, p.3393, 2019.
, Polymorphisms in inflammasome' genes and susceptibility to HIV-1 infection, J. Acquir. Immune Defic. Syndr, vol.59, pp.121-125, 2012.
, Susceptibility to Mycobacterium tuberculosis infection in HIV-positive patients is associated with CARD8 genetic variant, 2013.
, J. Acquir. Immune Defic. Syndr, vol.63, pp.147-151
, Deubiquitination of NLRP3 by BRCC3 critically regulates inflammasome activity, 2013.
, Mol. Cell, vol.49, pp.331-338
, Regulation of CDC42 GTPase by proline-rich tyrosine kinase 2 interacting with PSGAP, a novel pleckstrin homology and Src homology 3 domain containing rhoGAP protein, J. Cell Biol, vol.152, pp.971-984, 2001.
, HMGB1-dependent triggering of HIV-1 replication and persistence in dendritic cells as a consequence of NK-DC cross-talk, PLoS ONE, vol.3, p.3601, 2008.
, Src-dependent autophagic degradation of Ret in FAK-signalling-defective cancer cells, EMBO Rep, vol.13, pp.733-740, 2012.
, Extracellular ATP acts on P2Y2 purinergic receptors to facilitate HIV-1 infection, J. Exp. Med, vol.208, pp.1823-1834, 2011.
, The P2Y2 nucleotide receptor mediates vascular cell adhesion molecule-1 expression through interaction with VEGF receptor-2 (KDR/Flk-1), J. Biol. Chem, vol.279, pp.35679-35686, 2004.
, Monosodium urate crystals in inflammation and immunity, Immunol. Rev, vol.233, pp.203-217, 2010.
, How HIV takes advantage of the cytoskeleton in entry and replication, Viruses, vol.3, pp.293-311, 2011.
, P2X-selective purinergic antagonists are strong inhibitors of HIV-1 fusion during both cell-to-cell and cell-free infection, J. Virol, vol.88, pp.11504-11515, 2014.
, Interactions between HIV-1 and the cell-autonomous innate immune system, Cell Host Microbe, vol.16, pp.10-18, 2014.
, The NLRP3 inflammasome instigates obesity-induced inflammation and insulin resistance, Nat. Med, vol.17, pp.179-188, 2011.
, Negative regulation of the NLRP3 inflammasome by A20 protects against arthritis, Nature, vol.512, pp.69-73, 2014.
, LIM kinase 1 modulates cortical actin and CXCR4 cycling and is activated by HIV-1 to initiate viral infection, J. Biol. Chem, vol.286, pp.12554-12564, 2011.
, Rapid inflammasome activation in microglia contributes to brain disease in HIV/AIDS, Retrovirology, vol.11, p.35, 2014.
, Regulation of the formation of osteoclastic actin rings by proline-rich tyrosine kinase 2 interacting with gelsolin, J. Cell Biol, vol.160, pp.565-575, 2003.
, HIV envelope-CXCR4 signaling activates cofilin to overcome cortical actin restriction in resting CD4 T cells, Cell, vol.134, pp.782-792, 2008.
, At 72 hours post-CBL siRNA transfection MDMs were infected with HIV-1 AD8 (MOI of 1) for 6 hours, cell lysates were assayed for protein expression by western blot. For short hairpin RNA (shRNA) lentiviral particles transduction, the pLKO.1 shRNA expression lentiviral vector coding for each targeted gene was purchased from Thermo Scientific. The shRNAs used in knockdown experiments had the following sequences for P2Y2 (shRNA1, 5 0 -ATGTTCCACCTGGCTGTGTCTGATGCACT-3 0 ), for NLRP3 (shRNA1, 5 0 -TTCTTGAAGTGTTTCTAACGC-3 0 , shRNA2, 5 0 -AAACAGTAGAACAATTCCAGC-3 0 ) and pLKO.1 empty vector control. Lentiviral vector particles were generated by cotransfection of three plasmids coding for the gag-pol HIV-1 genes (pCMV-HIV-1-GAG-POL), for the vector genome carrying shRNA of interest (pLKO.1 shRNA) and for the plasmid coding for an envelope of VSVG (pMDG-VSV-G). Co-transfection was performed in HEK293T cells using Fugene transfection reagent (Promega) according to the manufacturer's protocol. Two days after transfection, supernatants were filtered using 0.45-mm cellulose acetate filters, HEK293T cells are siGenome (Dharmacon) smart pool selected composed of a pool of four siRNAs having the following sequences for CBL, 2013.
, Viral stocks were standardized by quantification of CAp24 antigen in cell culture supernatants with an enzyme-linked immunoabsorbent assay (Perkin Elmer) and infection of TZM-bl cells (bearing the b-galactosidase gene under the control of HIV-1 LTR) with serial dilutions of the viral stocks followed by cell fixation and X-Gal staining. The multiplicity of infection (MOI) was determined at 48 hours of infection of TZM-bl based on the number of the positive X-Gal cells. Viral infections After 3 days of infection with HIV-1 NDK (MOI of 1), PHA/IL-2-stimulated peripheral blood lymphocytes were cocultured with uninfected lymphoblasts or alone for 48 hours and analyzed by immunofluorescence for synapse formation. MDMs were infected with HIV-1 BaL (with a MOI of 2) during 3 days for the analysis by Proximity Ligation Assay (PLA) (following manufacturer's instructions) and the intracellular CAp24 by FACS, as previously described (Sé ror et al., 2011), for MSU (100 mM) and LPS (10 ng/ml) treated cells. MDMs or PMA-THP1 macrophages infected with HIV-1 AD8 were pre-treated 18 hours with MSU (100 mM) and AR-C118925XX (100 mM) and infected during six hours with HIV-1 AD8 (MOI of 1) in the presence of the indicated drugs. The quantification of the LDH release in the cell supernatants of MSU-treated MDM was performed using the commercially available ELISA kits for LDH (Roche) according to the manufacturers' instructions at 24 hours after infection. THP1 cells were also infected with HIV-1 NL4-3 (MOI of 1) or HIV-1 NL4-3DEnv (MOI of 1) during 6 hours and analyzed for related signaling pathways by western blot, Cells or THP1 cells (1x10 stocks of wild-type HIV-1 NL4-3 or HIV-1 AD8 , HEK293T cells (2x10 6 ) were transfected with 20 mg of the corresponding proviral DNA (pHIV-1 NL4-3 or pHIV-1 AD8 ) and for Env-deleted VSV-G pseudotyped NL4-3 viruses (pHIV-1 NL4-3DEnv ), HEK293T cells (2x10 6 ) cells were transfected with 4 mg of VSV-G expression vector, 2004.
, MOI of 1) at 48 hours after shRNA transduction for 6 hours and after washings were suspended in equal medium volumes. Cell supernatants were then harvested at 6 and 12 days after infection for quantification of HIV-1 CAp24 contents. For immunoprecipitation assays, THP1 cells were treated 8 hours with PP1 (20 mM), PP2 (20 mM) or 2 hours of pretreatment with UTP (100 mM) before infection. Then, THP1 cells were infected during 6 hours with HIV-1 NL4-3 (MOI of 1) in the presence of the indicated drugs. To remove the membrane bound non-internalized HIV-1 particles before analysis of intracellular CAp24 by western blot, cells were treated with trypsin at 37 C for 20 minutes followed by two extensive washings. Target cell infectivity was evaluated using the enhanced b-galactosidase assay kit (Roche), For THP1 monocytes or CEM-SS T cells silenced for NLRP3, through the transduction of lentiviral vectors expressing shRNA1 and shRNA2 against NLRP3 gene, and control cells were infected with HIV-1 NL4-3 or HIV-1 AD8
, After paraffin removal, slides were subjected to antigen retrieval by microwave boiling in 1 mmol/l EDTA pH 9.0. Cell slides were then incubated during overnight for immunofluorescence with anti-P2Y2 (Alomone), anti-NLRP3 (Adipogen), anti-CD163 (BD laboratories) or anti-gp120 (2G12) (AIDS Research and Reference Reagent Program, Division of AIDS, NIAID) or for immunochemistry with anti-NLRP3 (Sigma) after permeabilization in 0.3% Triton for 5 minutes and saturation in PBS-FBS 20% for 1 hour. Then, cells were incubated with appropriate secondary antibodies conjugated to Alexa Fluor 488, 546 or 647 fluorochromes (Life technologies) at room temperature during 1 hour and 30 minutes. Actin was stained with Alexa Fluor 594 Phalloidin for 30 minutes at room temperature, PBS or 0.1% Triton for MDMs and PBLs, and incubated with PBS-FBS 20% for 1 hour, vol.33342
, Adipogen) and anti P2Y2 (Alomone) as primary antibodies. Images of cells were acquired by laser-scanning fluorescent confocal microscopy Leica SPE with LAS-X software (Leica) with 8 bits configuration and using a 63X oil objective (1.4 numerical aperture). Leica Microsystem immersion oil (11513859) was used and imaging was performed at room temperature. Confocal images were analyzed with ImageJ and exported as TIFs for figures illustration. For 3D immunofluorescence intensity image construction, Proximity Ligation Assay (DUOLINKâ, Sigma) was performed according to the manufacturer's instructions using anti-NLRP3
, Antibody anti-NLRP3 was from Adipogen (Cryo-2), anti-P2Y2 was from Alomone, anti-ubiquitin (UB) (P4D1) was from Santa-Cruz, and anti-Flag was from Sigma. For immunoprecipitations, cell pellets were lysed at indicated times after infection or 24 hours after transfection in CHAPS buffer (50 mM Tris-HCl (pH = 7.5), 0.50 M NaCl and 0.1% CHAPS) containing protease and phosphatase inhibitors. Anti-P2Y2 (Alomone) antibodies were incubated overnight at 4 C with 500 mg cell lysates. The complexes were precipitated by incubation with Protein G Sepharose 4 Fast Flow (GE Healthcare) for 4 hours. For relative expression quantification analyzed protein bands were quantified by ImageJ software and normalized on the corresponding endogenous expression of GAPDH or ACTIN proteins, Western blots and immunoprecipitations Cells were washed twice with PBS and lysed in appropriated buffer (250 mM NaCl, 0.1% NP-40, 5 mM EDTA, 10 mM Na3VO4, 10 mM NaF, 5 mM DTT, 3 mM Na4P2O7, 1 mM EGTA, 10 mM Glycerol phosphate, 10 mM Tris-Hcl (pH = 7.5) and the protease and phosphatase inhibitors (Roche)). 10-40 mg of protein extracts were run on 4%-12% or 10% SDS-PAGE and transferred at 4 C onto a nitrocellulose membrane (0.2 Micron), vol.1, p.206
, Cell Reports, vol.28, pp.1-7, 2019.