Journal: Alzheimer's & Dementia

Article Title: Female‐biased astrocytic priming shapes early locus coeruleus vulnerability in an Aβ oligomer milieu

doi: 10.1002/alz.71168

Figure Lengend Snippet: Astrocyte‐linked C3 elevation and lactate–MCT2–OXPHOS axis engagement in the pons of female APP/PS1 mice. Astrocyte‐enriched pons fraction shows increased C3 expression in 2‐ to 3‐month‐old female APP/PS1 mice. A, Schematic illustrating MACS‐based isolation of pontine astrocytes. Schematic created using Biorender. B, Assessment of C3 expression by qPCR followed by 1% agarose gel electrophoresis–gel image showing C3 bands at 172 bp in astrocyte‐enriched fractions (lane 1–4) and astrocyte‐depleted fractions (lane 5–8). C, D, Bar graphs demonstrating elevated C3 levels in the astrocyte‐enriched fraction of female APP/PS1 mice compared to other groups and to their corresponding negative fraction. Each bar represents pooled pons samples from 8 mice per group. Lactate–MCT2 axis and OXPHOS pathway engagement in female APP/PS1 mice. Female APP/PS1 mice showed higher (E) lactate levels in the pons measured by colorimetric assay and increased (F) MCT2 expression assessed by qPCR ( n = 6 per group; two‐way ANOVA with Tukey post hoc test; data shown as mean ± SEM; * p < 0.05, ** p < 0.01). G, H, mRNA expression profiling of mitochondrial markers by qPCR in the pons. Female APP/PS1 mice showed significantly elevated expression of the mitochondrial complex I subunit Ndufs8 and complex V subunit Atp5a1 , indicating adaptive mitochondrial activation in the pons ( n = 5‐6; two‐way ANOVA with Tukey post hoc test; data shown as mean ± SEM; * p < 0.05, ** p < 0.01). I, Graphical summary. Elevated Aβ oligomers in female APP/PS1 mice shift astrocytes from a resting to an active GFAP +ve state, with associated C3 and NF‐κB2 upregulation and engagement of lactate–OXPHOS axis, supporting increased energetic demands. Graphical summary created using Biorender. Aβ, amyloid beta; ACSA‐1, astrocyte cell surface antigen 1; ANOVA, analysis of variance; Atp5a1, ATP synthase F1 subunit α; C3, complement component 3; GLUT1, glucose transporter 1; MACS, magnetic‐activated cell sorting; MCT2, monocarboxylate transporter 2; Ndufs8, NADH dehydrogenase iron‐sulfur protein 8; NF‐κB2, nuclear factor‐kappa‐B p100/p52 subunit; OXPHOS, oxidative phosphorylation; qPCR, quantitative polymerase chain reaction; SEM, standard error of mean; WT, wild type.

Article Snippet: Cells were incubated with FcR blocking reagent (Miltenyi Biotec, 130‐092‐575) at a 1:9 dilution for 10 minutes to prevent non‐specific antibody binding, followed by incubation with Anti‐GLAST or Astrocyte Cell Surface Antigen‐1 (ACSA‐1)‐Biotin (Miltenyi Biotec, 130‐095‐826) for 10 minutes.

Techniques: Expressing, Isolation, Agarose Gel Electrophoresis, Colorimetric Assay, Activation Assay, FACS, Phospho-proteomics, Real-time Polymerase Chain Reaction

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: HBVZ10, an AAV8 vector-based new HBV therapy candidate for cccDNA elimination

doi: 10.1016/j.omtm.2025.101646

Figure Lengend Snippet: Progressive reduction of both serum HBeAg and HBsAg upon blocking of cccDNA replenishment (A) Experimental procedures and time course of HBV infection and treatment evaluation using uPA/SCID chimeric mice model. (B1) The expressed anti-HBs antibody levels were >100,000 mIU/mL and sustained at steady levels for 200 days after a single injection of HBVZ10 in 8 chimeric mice (T1 and CT1-CT7) infected with HBV. A pretreatment serum sample from each replicate was used as a negative control, and all pretreatment anti-HBs antibody levels were plotted as 1 mIU/mL for consistency. (B2) Average anti-HBs antibody levels per group. HBVZ10 was administered on day 11 with a dose of 2.5E10 copies for mice CT1-CT4 on day 22 with 1.8E11 copies for mice T1 and CT5-CT6, or on day 44pi with 7E11 copies of HBV10 for mouse CT7. All 7 mice (CT1-CT7) also received 12-week ETV 10 days after HBVZ10 injection. T. HBVZ10 monotherapy. CT: Combination of HBVZ10 with ETV. (C1) Serum HBeAg kinetics among 7 mock-treated mice (MT1-MT7 blue) and 8 treated mice (T1 and CT1-CT7 green). Serum HBeAg levels are expressed as the OD ratio of the tested sample to the mean OD of three negative controls. A ratio greater than 2.1 is considered positive. (C2) Average serum HBeAg levels per group between mock-treated (blue) and treated (green) from B1. (D1) Kinetic serum HBsAg levels (IU/mL) among mock-treated 7 mice (blue) and 8 treated mice (green) from B1. (D2) Average serum HBsAg levels per group between mock-treated (blue) and treated group (green) from C1. (E) Comparison of average cccDNA levels between mock-treated and combination treated groups (cccDNA was undetectable in mice CT2 and CT4 and not plotted). (F) Comparison of average rcDNA levels between mock-treated and combination-treated groups. Statistical significance was determined using Student’s t test; p < 0.05 was considered significant. Error bars: standard deviation (SD).

Article Snippet: Blood was collected triweekly for quantification of serum HBV DNA (qPCR), HBeAg (CSB-E13557h, CUSABIO), HBsAg (GS HBsAg EIA 32591, Bio-Rad), anti-HBs antibody (MONOLISA anti-HBs EIA 25200, Bio-Rad) with calibrators (MONOLISA anti-HBs 20-Calibrator kit 25219, Bio-Rad), and human albumin (human albumin ELISA kit E−80AL, Immunology Consultants Laboratory) levels by ELISA per instructions.

Techniques: Blocking Assay, Infection, Injection, Negative Control, Comparison, Standard Deviation

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: HBVZ10, an AAV8 vector-based new HBV therapy candidate for cccDNA elimination

doi: 10.1016/j.omtm.2025.101646

Figure Lengend Snippet: Add-on of HBVZ10/anti-HBs antibodies to 9-week ETV treatment prevented HBV relapse, leading to progressive reduction of both serum HBeAg and HBsAg levels (A) Experimental procedures and time course of HBV infection and treatment evaluation using uPA/SCID chimeric mice model. (B) Serum HBV DNA (copies/mL log10) kinetics in mock-treated (MT1-MT8 blue) and ETV monotherapy (ET1-ET4 orange) group. (C) Serum HBV DNA kinetics in 8 ETV-treated mice (CT1-CT8) with add-on with a single dose of 1E10 copies of HBVZ10 on day 70pi, then anti-HBs levels were further boosted with administration of mouse anti-HBs at a dose of 250 μg/injection triweekly started on day 99 or later. (D) Average serum HBV DNA levels (copies/mL log10) among 3 groups. (E1) Serum kinetic anti-HBs antibody levels (mIU/mL) expressed by HBVZ10 and boosted by infusions of mouse anti-HBs antibody among the 8 mice (CT1-CT8). Dashed line: 100,000 mIU/mL. Note: Anti-HBs antibody levels exhibited significant fluctuations in some mice receiving triweekly infusions of mouse anti-HBs antibody, contrasting the steady and consistent antibody levels expressed by sufficient HBVZ10 doses ( A). A pretreatment serum sample from each replicate was used as a negative control, and all pretreatment anti-HBs antibody levels were plotted as 1 mIU/mL for consistency. (E2) Average anti-HBs antibody levels (mIU/mL) per group. (F1) Kinetic serum HBsAg levels (IU/mL) among 2 mice with mock-treated (MT1 and MT2 blue), 2 mice with 9-week of ETV therapy (ET1 and ET2 yellow), and 8 mice with 9-week of ETV and anti-HBs antibodies add-on (CT1-CT8 green). (F2) Average serum HBsAg levels (IU/mL) per group. (G1) Kinetic serum HBeAg levels among 2 mice with mock-treated (MT1 and MT2 blue), 2 mice with 9-week of ETV therapy (ET1 and ET2 yellow), and 8 mice with 9-week of ETV and anti-HBs antibodies add-on (CT1-CT8 green). Note. Mouse CT5 had relatively higher baseline HBeAg level and HBeAg remained detectable on termination day (day 218 PI) despite progressive reduction. Serum HBeAg levels are expressed as the OD ratio of the tested sample to the mean OD of three negative controls. A ratio greater than 2.1 is considered positive. (G2) Average serum HBeAg levels per group. (H and I) Comparison of intracellular cccDNA and rcDNA levels among 2 livers with 9-week of ETV therapy (ET1 and ET2 yellow), 2 livers with mock-treated (MT1 and MT2 blue), and 6 livers with 9-week of ETV and anti-HBs antibodies add-on, 5 (CT1-CT3, CT7 and CT8) of them achieved progressive reduction of both serum HBeAg and HBsAg to undetectable levels and the remaining one with detectable serum HBeAg on the termination day (CT5). The differences in cccDNA and rcDNA levels were analyzed by Student t test, p < 0.05 considered significant. Error bars: standard deviations (SD). The limit of detection for HBV DNA is 100 copies/mL and 0.05 IU/mL for serum HBsAg.MT: mock-treated. ET: ETV Monotherapy. CT: 9-week of ETV and anti-HBs antibodies add-on.

Article Snippet: Blood was collected triweekly for quantification of serum HBV DNA (qPCR), HBeAg (CSB-E13557h, CUSABIO), HBsAg (GS HBsAg EIA 32591, Bio-Rad), anti-HBs antibody (MONOLISA anti-HBs EIA 25200, Bio-Rad) with calibrators (MONOLISA anti-HBs 20-Calibrator kit 25219, Bio-Rad), and human albumin (human albumin ELISA kit E−80AL, Immunology Consultants Laboratory) levels by ELISA per instructions.

Techniques: Infection, Injection, Negative Control, Comparison

Journal: Retrovirology

Article Title: Intra- pol proviral open region of HTLV-1 controls the transcription from both long terminal repeats

doi: 10.1186/s12977-025-00671-4

Figure Lengend Snippet: Repressive effect of IPOR on viral production and transcription from the integrated provirus. A Diagram illustrating the experimental workflow for quantitative determination of HTLV-1 p19 and measurement of tax expression using HTLV-1-WT or Mut molecular clones. B Production of viral-protein p19 measured with ELISA method. C Relative mRNA expression of tax in HTLV-1-WT- or Mut- infected JET cells. ΔΔCt correcting values ( tax /18S rRNA) by RT-qPCR were adjusted to PVL. At least two independent experiments were performed. Technical triplicate data is presented as mean ± SD. p values were calculated by Welch’s t test

Article Snippet: After 48 h, p19 levels in the virus-containing supernatants were quantified using the RETRO-TEK HTLV p19 Antigen ELISA Kit (ZeptoMetrix).

Techniques: Expressing, Clone Assay, Enzyme-linked Immunosorbent Assay, Infection, Quantitative RT-PCR

Journal: Nature Communications

Article Title: The Human T-cell Leukemia Virus capsid protein is a potential drug target

doi: 10.1038/s41467-025-65899-2

Figure Lengend Snippet: a Graph comparing the infectivity of WT-A and WT-C packaging constructs as measured by GFP reporter expression in a replication dependent infection system. Results are displayed from 8 independent experiments. Statistical analysis was performed using an unpaired two-tailed t-test. Bar graph displaying infectivity of HIV-1 ( b ) and HTLV-1c ( c ) as measured by GFP reporter expression in a replication dependent infection system in the presence of increasing amounts of lenacapavir. Results are displayed as the mean +/− SD from 4 independent experiments. Statistical analysis was performed using an unpaired two-tailed t-test. d Bar graph displaying relative infectivity (grey bars) and p19 production (white bars) of 21 HTLV-1c CA mutants, compared to the WT construct, which is set at 100%. Results are displayed as the mean +/- SD from 3 independent experiments. Shaded areas contain residues in the CA intra-hexamer interface (pale green, e ), CA NTD trimeric interface (pale purple, f ), SO 4/ PO 4 binding pocket (pale orange, g ) and CA CTD trimer of dimers interface (pale blue, h ). i HTLV-1c CA mutants, shown in ( d ), mapped onto the structure of the monomeric full-length CA protein, with mutated residues coloured according to infection and virus production phenotypes. Representative gating strategy for the FACS data is provided in Supplementary Fig. . Source data are provided as a Source Data file.

Article Snippet: Virus production was measured using a p19 antigen ELISA kit (Zeptometrix) according to the manufacturer’s instructions.

Techniques: Infection, Construct, Expressing, Two Tailed Test, Binding Assay, Virus

Journal: Nature Communications

Article Title: Identification of inducible HIV reservoirs in tonsillar, intestinal and cervical tissue models of HIV latency

doi: 10.1038/s41467-025-65288-9

Figure Lengend Snippet: a Representation of the experimental workflow for studying productive and latent HIV infection in tonsillar (TO), intestinal (GUT) and cervicovaginal (CVX) tissues. Conditions: uninfected (HIV-), infected (HIV+), ART-treated (ART), and ART-treated LRA-stimulated (ART+LRA). Created with BioRender: Genescà, M. (2025) https://BioRender.com/wd0r7zx . b Productive HIV infection in the tissue models. Graphs display the percentages of p24 + cells in CD4 + T cells from uninfected and infected tonsils (day 5), and intestinal and cervical tissues (day 6). N = 23, 26 and 5 biological replicates for tonsils, intestine and cervix. Means are represented. c Longitudinal monitoring of HIV infection in tonsillar and intestinal tissue models. Graphs show the percentage of p24 + cells in CD4 + T cells under uninfected, infected and infected ART-treated conditions over 8-9 days. N = 6 biological replicates per tissue. Means and standard error of the means (SEM) are shown. d Effect of ART on HIV infection in tissue models. Graphs display the frequencies of p24 + cells in CD4⁺ T cells from HIV-infected tissues with or without ART for 2–3 days. N = 12, 13 and 7 biological replicates for tonsils, intestine and cervix. Means are represented. e Impact of HIV infection and ART on CD4 + T cells. Graphs depict the frequencies of CD4 + T cells under uninfected, infected and infected ART-treated conditions in all tissue models at days 7-9. N = 6, 6 and 4 biological replicates for tonsils, intestine and cervix. Means and SEM are depicted. f Viral reactivation of the inducible reservoir in the tissue models. Graphs show the frequencies of p24 + cells in CD4 + T cells isolated from HIV-infected ART-treated tissue blocks either unstimulated or stimulated with phorbol 12-myristate 13-acetate and ionomycin (PMA+Iono). N = 12, 13 and 11 biological replicates for tonsils, intestine and cervix. Means are represented. Statistical comparisons between conditions were performed using the Wilcoxon test ( b , c , d , f ) and the Friedman test ( c , e ), both two-sided. Significance levels are indicated numerically. All experiments were independently replicated, each run including tissue samples from distinct donors. Source data are provided as a Source Data file.

Article Snippet: HIV-1 Gag p24 concentrations in the supernatants were quantified using the Simoa® HIV p24 Advantage Kit (Quanterix, USA; cat. no. 102215) on a Simoa HD-1 analyzer, following the manufacturer’s instructions.

Techniques: Infection, Isolation

Journal: Nature Communications

Article Title: Identification of inducible HIV reservoirs in tonsillar, intestinal and cervical tissue models of HIV latency

doi: 10.1038/s41467-025-65288-9

Figure Lengend Snippet: a Opt-SNE plots displaying the distribution of the identified CD4 + T-cell clusters from uninfected tonsils (TO,T) at day 8, and intestines (GUT,G) and cervix (CVX,C) at day 9. ( b ) Heatmap illustrating the Mean Fluorescence Intensity (MFI) of the phenotypic markers across the clusters. c Stacked bar charts showing the median percentage of each CD4 + T-cell cluster from the uninfected tissues. Statistical comparisons were performed using two-sided Kruskal-Wallis tests, with Dunn’s post hoc correction, and significance indicated (* p < 0.05; ** p < 0.01, *** p < 0.001; **** p < 0.0001). d Pie charts representing pairwise comparisons of cluster proportions between the uninfected tissues. Median percentages are shown. Statistical values derived from the edgeR method (two-sided), with significance levels indicated (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). e Volcano plots showing the differential cluster proportion between the uninfected (HIV-) and infected (HIV+) conditions in tonsillar and intestinal tissues. The edgeR method (two-sided) was employed, with green dots indicating statistically significant differences (p < 0.05). f HIV infection levels in CD4 + T-cell clusters. Graphs illustrating the percentage of p24 + cells within each cluster from tonsillar (day 8) and intestinal (day 9) tissues. Percentages in HIV- were subtracted. Medians and quartiles are represented. Statistical comparisons between infected and uninfected conditions were performed using a two-sided Wilcoxon test. g Pie charts depicting the contribution of each cluster to total HIV-infected CD4 + T cells from tonsillar and intestinal tissues. Percentages were calculated as the proportion of p24⁺ cells in each cluster relative to total p24⁺ CD4⁺ T cells. Medians are shown, and statistical comparisons between tissues were performed using a two-sided Mann-Whitney test (** p < 0.01; **** p < 0.0001). Smaller pie charts represent the median percentage of each cluster within the total CD4 + T-cell population. N = 9 biological replicates for tonsils and intestine, N = 10 for cervix in A-D. All experiments were independently replicated, each run including tissue samples from distinct donors. Source data are provided as a Source Data file.

Article Snippet: HIV-1 Gag p24 concentrations in the supernatants were quantified using the Simoa® HIV p24 Advantage Kit (Quanterix, USA; cat. no. 102215) on a Simoa HD-1 analyzer, following the manufacturer’s instructions.

Techniques: Fluorescence, Derivative Assay, Infection, MANN-WHITNEY

Journal: Nature Communications

Article Title: Identification of inducible HIV reservoirs in tonsillar, intestinal and cervical tissue models of HIV latency

doi: 10.1038/s41467-025-65288-9

Figure Lengend Snippet: a Inducible HIV reservoirs in CD4 + T-cell clusters after viral reactivation with PMA and ionomycin (PMA+Iono). Graphs show the percentage of p24 + cells within each cluster of virally reactivated CD4 + T cells from tonsillar tissues (TO) at day 8 and intestinal tissues (GUT) at day 9. Percentages in the ART-treated condition were subtracted from those in the PMA/ionomycin-stimulated condition. Median values with quartiles are represented. Statistical comparisons between the unstimulated (ART) and PMA/ionomycin-reactivated conditions were conducted using the two-sided Wilcoxon test, with significance indicated numerically. b Pie charts illustrating the contribution of each cluster to the overall inducible reservoir in CD4 + T cells from tonsillar and intestinal tissues. Percentages were calculated as the proportion of p24⁺ cells in each cluster relative to total p24⁺ CD4⁺ T cells. Medians of these percentages are depicted, and statistical comparisons were performed using the two-sided Mann-Whitney test, with significance levels denoted (* p < 0.05; ** p < 0.01 and *** p < 0.001). Smaller pie charts represent the median percentage of each cluster within the CD4 + T-cell population under PMA/ionomycin-stimulated condition. c Pie charts displaying the distribution of clusters derived from Boolean gating of phenotypic markers within the p24 + CD4 + T-cell population. Arcs represent the expression of the phenotypic markers, each depicted in a distinct colour. All p24 + CD4 + T cells express CD69. N = 9 biological replicates each for tonsils and intestine. All experiments were independently replicated, each run including tissue samples from distinct donors. Source data are provided as a Source Data file.

Article Snippet: HIV-1 Gag p24 concentrations in the supernatants were quantified using the Simoa® HIV p24 Advantage Kit (Quanterix, USA; cat. no. 102215) on a Simoa HD-1 analyzer, following the manufacturer’s instructions.

Techniques: MANN-WHITNEY, Derivative Assay, Expressing

Journal: Nature Communications

Article Title: Identification of inducible HIV reservoirs in tonsillar, intestinal and cervical tissue models of HIV latency

doi: 10.1038/s41467-025-65288-9

Figure Lengend Snippet: a Viral reactivation in ART-treated CD4 + T cells from tonsillar (TO), intestinal (GUT) and cervical (CVX) tissues. Graphs display the percentage of p24 + cells within CD4 + T cells in unstimulated (ART) and LRA-stimulated conditions: PMA and ionomycin (PMA+Iono), Ingenol (ING), Romidepsin (RMD), Ingenol and Romidepsin (I+R), Panobinostat (PNB), AZD5582 (AZD) and IL-15. N = 12, 13 and 11 biological replicates for tonsils, intestine and cervix. Percentages in ART were subtracted from those in each LRA-stimulated condition. Medians with quartiles are represented. Statistical comparisons between unstimulated and LRA-reactivated conditions were conducted. b Pie charts illustrating the contribution of LRAs to HIV reactivation in tonsillar and intestinal CD4 + T cells. N = 9 and 7 biological replicates for tonsils and intestine. Means are depicted. Statistical comparisons between tissues were performed. c SIMOA measurements of p24 in supernatants from tonsillar and intestinal CD4⁺ T cells HIV-infected (HIV+), ART-treated and ART-treated LRA-stimulated. N = 3 biological replicates per tissue. d Effect of LRAs on ART-treated tonsillar and intestinal CD4 + T-cell subpopulations. Graphs display the percentage of p24 + cells within clusters following stimulation with ING, I+R, AZD or IL-15. Percentages in ART were subtracted. N = 9 biological replicates per tissue. Medians with quartiles are represented. Statistical comparisons between unstimulated and LRA-reactivated conditions were conducted. e Pie charts illustrating the contribution of each cluster to total reactivated CD4 + T cells from tonsillar and intestinal tissues. Percentages were calculated as the proportion of p24⁺ cells in each cluster relative to total p24⁺ CD4⁺ T cells. N = 9 biological replicates per tissue. Median percentages are depicted, and statistical comparisons between tissues were conducted. Smaller pie charts represent the median percentage of each cluster within the LRA-stimulated CD4 + T-cell population. Statistical tests (all two-sided): Wilcoxon test ( a , d ); Mann-Whitney ( b , e ). Significance levels are indicated numerically or as * p < 0.05; ** p < 0.01; *** p < 0.001. All experiments except ( c ) were independently replicated, each run including tissue samples from distinct donors. Source data are provided as a Source Data file.

Article Snippet: HIV-1 Gag p24 concentrations in the supernatants were quantified using the Simoa® HIV p24 Advantage Kit (Quanterix, USA; cat. no. 102215) on a Simoa HD-1 analyzer, following the manufacturer’s instructions.

Techniques: Infection, MANN-WHITNEY

Journal: Nature Communications

Article Title: Identification of inducible HIV reservoirs in tonsillar, intestinal and cervical tissue models of HIV latency

doi: 10.1038/s41467-025-65288-9

Figure Lengend Snippet: Viral reactivation with LRAs in the ART-treated CD4 + T-cell clusters from tonsillar and intestinal tissues: a C03:T CM , b C06:T TEM CD69+ , c C08:T FH CD69+ CCR7+ , d C09:T FH CD69+ CCR7- , e C10:T RM CD69+ PD-1+ , f C11:T RM CD69+ CD49a+ , and g C12:T RM CD69+ CD49a+ CD103+ . Graphs displaying the percentage of p24 + cells within the CD4 + T-cell subpopulations in unstimulated (ART) and LRA-stimulated conditions: PMA and ionomycin (PMA+Iono), Ingenol (ING), Romidepsin (RMD), Ingenol and Romidepsin (I+R), Panobinostat (PNB), AZD5582 (AZD) and IL-15. Percentages of p24 + cells in ART were subtracted from those in each LRA-stimulated condition. N = 9 biological replicates per tissue. Medians with quartiles are represented. Statistical comparisons between unstimulated and LRA-reactivated conditions were conducted using the two-sided Wilcoxon test, with significance levels indicated numerically. All experiments were independently replicated, each run including tissue samples from distinct donors. Source data are provided as a Source Data file.

Article Snippet: HIV-1 Gag p24 concentrations in the supernatants were quantified using the Simoa® HIV p24 Advantage Kit (Quanterix, USA; cat. no. 102215) on a Simoa HD-1 analyzer, following the manufacturer’s instructions.

Techniques:

Journal: Nature Communications

Article Title: Identification of inducible HIV reservoirs in tonsillar, intestinal and cervical tissue models of HIV latency

doi: 10.1038/s41467-025-65288-9

Figure Lengend Snippet: Percentage of p24 + cells in sorted ( a ) tonsillar (TO) T NA (CD45RO⁻ CCR7⁺ CD69⁻) and T FH (CXCR5⁺ PD-1⁺) subpopulations and ( b ) intestinal (GUT) T RM (CD45RO⁺ CCR7⁻ CD69⁺ CXCR5⁻) and T NON-RM (CD69⁻) CD4⁺ T cells unstimulated (ART) and following stimulation with PMA and ionomycin (PMA+Iono), the combination of Ingenol and Romidepsin (I+R) and IL-15. Ingenol (ING) was also used for tonsillar cells. Percentages of p24 + cells in ART were subtracted from those in each LRA-stimulated condition. Means with standard error of means (SEM) are represented. c Effect of LRA stimulation on the expression of phenotypic markers in sorted CD4⁺ T-cell populations from tonsillar and intestinal tissues. Histograms represent the Mean Fluorescence Intensity (MFI) of each marker across conditions within the CD4 + T-cell subpopulations. N = 1 and 2 biological replicate for tonsils and intestine, respectively. Experiments with intestinal samples were duplicated in independent runs, each run including one tissue sample from a distinct donor. Source data are provided as a Source Data file.

Article Snippet: HIV-1 Gag p24 concentrations in the supernatants were quantified using the Simoa® HIV p24 Advantage Kit (Quanterix, USA; cat. no. 102215) on a Simoa HD-1 analyzer, following the manufacturer’s instructions.

Techniques: Expressing, Fluorescence, Marker