The anti-caspase 1 inhibitor VX-765 reduces immune activation, CD4+ T cell depletion, viral load, and total HIV-1 DNA in HIV-1 infected humanized mice
- Department of Infection and Immunity, Luxembourg Institute of Health, Luxembourg
- Department of Infectious Diseases, University of Liège, CHU de Liège, Belgium
Figures
Figure 1 with 3 supplements
Heatmaps reveal a biphasic inflammasome-related genes transcriptomic upregulation upon HIV-1JRCSF infection of huNSG mice.
Relative mRNA expression of the indicated genes measured by qPCR in hCD45+ cells isolated from bone marrow, lung, lymph nodes and spleen in HIV-1JRCF i.p. infected huNSG mice at day 0 (n=6), day 1 …
Figure 1—figure supplement 1
Kinetics of viremia and early CD4+ T cell loss during HIV-1 infection of huNSG mice.
(A) Plasma HIV-1 viral load measured by ddPCR (LOD: 235 copies/ml) and (B) human CD4+ T cells percentage among CD3+ cells measured by flow in HIV-1JRCF i.p. infected NSG humanized mice at day 0 …
Figure 1—figure supplement 2
HIV-1 disseminates within 3 days post-infection in huNSG mice.
Viral RNA in the spleen, lung, lymph nodes (LN), bone marrow (BM) and in the blood were measured by RT-PCR (Log HIV RNA copies per 50 ng of total hCD45+ cells RNA) in HIV-1JRCF i.p. infected NSG …
Figure 1—figure supplement 3
HIV-1 induction of inflammasome related genes expression in tissues.
Relative mRNA expression of the indicated genes measured by qPCR in hCD45+ cells isolated from bone marrow (BM), lymph nodes (LN) and spleen in HIV-1JRCF i.p. infected huNSG mice at day 0 (n=5), day …
Figure 2 with 1 supplement
IFI16 and AIM2 expression correlates with HIV-1 replication and pathogenesis.
IFI16 and AIM2 relative mRNA expression comparison between viremic (HIV POS, n=8) and aviremic (HIV ND, n=16) huNSG mice (A and D) in the bone marrow (BM), lymph nodes (LN), and the spleen (SP). …
Figure 2—figure supplement 1
ASC and CASP-1 expression correlates with HIV-1 replication and pathogenesis.
ASC and CASP-1 relative mRNA expression comparison between viremic (HIV POS, n=7) and aviremic (HIV ND, n=16) huNSG mice (A, D, G) in the bone marrow (BM), spleen (SP) and in the lymph nodes (LN). …
Figure 3 with 1 supplement
IL-18 is induced by HIV-1 replication and correlates inversely with CD4+ T cells percentage.
(A) Plasma cytokines concentration were measured using a multiplex U-PLEX Biomarker assay kit in the plasma of HIV-1JRCF i.p. infected huNSG mice at day 0 (n=5), day 3 (n=5), day 10 (n=5), and day …
Figure 3—figure supplement 1
IFN-γ and IP-10 are inducted by HIV-1 replication.
(A) Plasma cytokines levels comparison between viremic (HIV POS, n=8) and aviremic (HIV ND, n=11) huNSG mice. Correlations between plasma cytokines levels with corresponding plasma viral load (B) or …
Figure 4 with 2 supplements
Caspase-1 inhibition reduces circulating levels of IL-18.
Plasma cytokine concentrations were measured using a multiplex U-PLEX Biomarker assay kit in longitudinal plasma samples of HIV-1JRCF i.p. infected huNSG mice treated with VX-765 (VX-765, n=12) or …
Figure 4—figure supplement 1
Caspase-1 inhibition reduces levels of plasma cytokines and mRNA expression of inflamasomme genes.
Plasma cytokine concentrations (A to D) weremeasured using a multiplex U-PLEX Biomarker assay kit in longitudinal plasma samples of HIV-1JRCF i.p. infected huNSG mice treated with VX-765 (VX-765, …
Figure 4—figure supplement 2
VX-765 treatment prevents CD4+ T cell depletion in the blood.
Blood human CD4+ T cells percentage among CD3+ cells (A and B) and CD4+/CD8+ T cell ratios (C and D) measured longitudinally by flow cytometry in VX-765 (VX-765, n=12) or vehicle (Vehicle, n=12) …
Figure 5
VX-765 treatment prevents CD4+ T cell depletion in the blood and bone marrow (BM) during HIV-1 infection of huNSG mice.
Blood human CD4+ T cells percentage among CD3+ cells (A) and CD4+/CD8+ T cell ratios (B) measured longitudinally by flow cytometry in VX-765 (VX-765, n=12) or vehicle (Vehicle, n=12) huNSG mice at 3 …
Figure 6
VX-765 treatment reduces plasma viral load and total HIV-1 DNA after HIV-1 infection of huNSG mice.
Plasma viral load (A) was measured longitudinally by ddPCR in vehicle (Vehicle, n=12) (B) or in VX-765 (VX-765, n=12) (C) or in treated HIV-1 infected huNSG mice at 3 days, 11 days, and 22 days …
Figure 7 with 1 supplement
Representative histograms of FLICA staining in splenocytes from VX-765 and vehicle-treated HIV-1 infected huNSG mice.
Splenocytes were stained for active caspase-1 using FAM-FLICA reagent. Representative MFI of FAM-FLICA+ CD4+, CD8+, CD11c+, and CD14+ splenic cells in one HIV-1 infected huNSG mice treated with …
Figure 7—figure supplement 1
Gating strategy for FLICA analysis.
FAM-Flica staining was analysed in gated human CD4+, CD8+, CD14+ and CD11c+ cells using the anti-hu CD45, anti-hu CD4, anti-hu CD8, anti-hu CD11c, anti-hu CD14 antibodies.
Figure 8
VX-765 treatment reduces caspase-1 activation in splenic CD11c+ and CD14+ in HIV-1 infected huNSG mice.
Splenocytes were stained for active caspase-1 using FAM-FLICA reagent. Percentage and MFI of FAM-FLICA+ CD11c+ (A, C), CD14+ (B, D) splenic cells in vehicle or VX-765 treated HIV-1 infected huNSG …
Figure 9 with 2 supplements
VX-765 treatment does not affect caspase-1 activation in splenic CD4+ and CD8+ T cells in HIV-1 infected huNSG mice.
Splenocytes were stained for active caspase-1 using FAM-FLICA reagent. Percentage and MFI of FAM-FLICA+ CD4+ (A, C), CD8+ (B, D) splenic cells in vehicle or VX-765 treated HIV-1 infected huNSG mice …
Figure 9—figure supplement 1
VX-765 treatment does not affect caspase-1 activation in the bone marrow.
Cells from the bone marrow were stained for active caspase-1 using FAM-FLICA reagent. Percentage of FAM-FLICA+ CD11c+ (A), CD14+ (B), CD4+ (C), CD8+ (D) splenic cells in vehicle or VX-765 treated …
Figure 9—figure supplement 2
Representative dot plots of FLICA versus LIVE-DEAD staining in splenocytes from VX-765 and vehicle-treated HIV-1 infected huNSG mice.
Author response image 1
Mean of percentage (± SD) of human CD45+ cells in blood, Lymph node (LN), Bone Marrow (BM), lung and spleen of humanized mice before (day 0) and after HIV infection (day 3, 10 and 24).
Author response image 2
Mean percentage (± SD) of human CD4+ T cells in human CD45+CD3+ cells in blood and tissues of humanized mice before (D0) and after HIV infection (D1, D3, D10 and D24).
Author response image 3
Mean percentage (± SD) of human CD8+ T cells in human CD45+CD3+ cells in blood and tissues of humanized mice before (D0) and after HIV infection (D1, D3, D10 and D24).
Author response image 4
Representative dot plots of FLICA versus LIVE-DEAD staining among debris in splenocytes from vehicle treated (left panel) and VX-765 (right panel) HIV-1 infected huNSG mice.
Author response image 5
Combined expression of CD38, HLA-DR and PD-1 in splenocytes from vehicle-treated and VX-765-treated (right panel) HIV-1 infected huNSG mice.
Tables
Author response table 1
LOD of the Mesoscale U-PLEX biomarker assay kit for the different cytokines.
| U-PLEX Assay | LLOD(Sensitivity) | Dynamic Range |
|---|---|---|
| Human IFN-γ | 1.7 pg/mL | 1.7-17,000 pg/mL |
| Human IL-1β | 0.15 pg/mL | 0.15-3,800 pg/mL |
| Human IL-2 | 0.70 pg/mL | 0.70-1,900 pg/mL |
| Human IL-6 | 0.33 pg/mL | 0.33-2,000 pg/mL |
| Human IL-12p70 | 0.69 pg/mL | 0.69-5,3000 pg/mL |
| Human IL-18 | 0.5 pg/mL | 0.5-14,000 pg/mL |
| Human IP-10 | 0.49 pg/mL | 0.49-6,000 pg/mL |
| Human MIP-1β | 1.5 pg/mL | 1.5-1,600 pg/mL |
| Human TNF-α | 0.51 pg/mL | 0.51-3,700 pg/mL |
| Human TRIAL | 0.66 pg/mL | 0.66-10,000 pg/mL |
