Journal: Nature Microbiology

Article Title: Vaccine plus microbicide effective in preventing vaginal SIV transmission in macaques

doi: 10.1038/s41564-023-01353-7

Figure Lengend Snippet: a , Comparison of SAMT-247 non-treated/treated effector cell-mediated ADCC activity in the vaccine ( n = 18) and vaccine + SAMT-247 groups ( n = 20; P < 0.0001). b , Correlation of SAMT-247-induced ADCC activity with number of intravaginal challenges in the vaccine + SAMT-247 group ( n = 20; P = 0.024). c , d , Intracellular Granzyme B, perforin, IFN‐γ and TNF-α in macaque rectal mucosal ( n = 9) NKG2A + cells in the presence or absence of different stimuli. e , Macaque rectal mucosal NKp44 + IL-17 + cells in the presence or absence of different stimuli ( n = 9). f , Correlation of efferocytosis with number of intravaginal challenges in animals in the vaccine group ( n = 18; P = 0.01). g , h , Comparison of percentage of efferocytosis ( P < 0.0001) ( g ) and efferocytosis MFI ( P < 0.0001) ( h ) using week 14 CD14 + monocytes in all vaccinated animals ( n = 38). i , Correlation of SAMT-247-induced efferocytosis (SAMT-247-untreated efferocytosis subtracted from SAMT-247-treated efferocytosis) with number of intravaginal challenges in the vaccine + SAMT-247 group ( n = 20; P = 0.065). Data shown in a , c , d , e , g and h were analysed with the two-tailed Wilcoxon signed-rank test. Data shown in b , f and i were analysed with the two-tailed Spearman correlation test. Horizontal and vertical bars denote mean and standard deviation, respectively.

Article Snippet: CD14 + cells were isolated from cryopreserved PBMCs (10 × 10 6 cells) collected following pre-study and 2 weeks post last immunization (week 14) by using non-human primate CD14 MicroBeads (#130-091-097, Miltenyi Biotec) following manufacturer instructions.

Techniques: Comparison, Activity Assay, Two Tailed Test, Standard Deviation

Journal: Nature Microbiology

Article Title: Vaccine plus microbicide effective in preventing vaginal SIV transmission in macaques

doi: 10.1038/s41564-023-01353-7

Figure Lengend Snippet: a, b) Intracellular Granzyme B, perforin, IFN‐γ, and TNF-α in healthy human (n = 6) blood NKG2A + cells in the presence or absence of different stimuli. c ) Comparison of Env-specific rectal NKp44 + IL-17 + cells between vaccine+SAMT-247 (n = 20) and vaccine group (n = 18) 1 week post last vaccination ( P = 0.43). d ) Correlation of rectal mucosal Env-specific NKp44 + IL-17 + cells with number of intra-vaginal challenges in the vaccine group (n = 18). e ) Gating of NKG2A + NK cells, NKp44 + ILCs, and NKG2A – NKp44 – ILCs in rectal mucosal samples in the presence of PMA or PMA + SAMT-247 at 12 hours post stimulation. Gating was done on singlets, live, CD45 + , CD3 − , CD20 − , CD11b − cells. f ) Gating of NKp44 + IL-17 + ILCs in the rectal mucosal sample in the presence of PMA or PMA + SAMT-247 at 12 hours post stimulation. g ) Correlation of efferocytosis percentage with number of intra-vaginal challenges in the vaccine+SAMT-247 group (n = 20). ( h-i ) Comparison of h ) percentage of efferocytosis ( P < 0.0001) and i ) efferocytosis MFI ( P < 0.0001) using pre CD14 + monocytes in all vaccinated animals (n = 38). Data shown in ( a, b, h, i ) were analyzed with the two-tailed Wilcoxon signed-rank test or two-tailed Mann-Whitney test. Data shown in ( d, g ) were analyzed with the two-tailed Spearman correlation test.

Article Snippet: CD14 + cells were isolated from cryopreserved PBMCs (10 × 10 6 cells) collected following pre-study and 2 weeks post last immunization (week 14) by using non-human primate CD14 MicroBeads (#130-091-097, Miltenyi Biotec) following manufacturer instructions.

Techniques: Comparison, Two Tailed Test, MANN-WHITNEY

Journal: Nature Microbiology

Article Title: Vaccine plus microbicide effective in preventing vaginal SIV transmission in macaques

doi: 10.1038/s41564-023-01353-7

Figure Lengend Snippet: a , Representative imaging of human NKG2A + cells unstimulated or stimulated with SAMT-247, PMA or PMA + SAMT-247. b , Mean zinc intensity in NKG2A + cells of the healthy human donor in the presence or absence of zinc chelator in different stimulation conditions ( n = 8). Fluorescence intensity of each field was measured for zinc expression as indicated by green colour, and the total number of DAPI positive cells were counted to determine the mean intensity of zinc/cells using iMARIS software. The mean of two duplicate fields was evaluated for the calculation. c , Comparison of expressions of NKG2A marker in macaques in the absence or presence of zinc chelator and stimuli in the vaccine + SAMT-247 group ( n = 4) and vaccine group ( n = 2). d – g , Comparison of expressions of granzyme B, perforin, IFN‐γ and TNF-α by macaque blood NKG2A + cells from week 17 in the absence or presence of different stimulations and zinc chelator in the vaccine + SAMT-247 group ( n = 4) and vaccine group ( n = 2). h , i , Evaluation of the frequency of CD14 + monocytes and CD14 + IL-10 + monocytes in the absence or presence of zinc chelator and stimuli in the vaccine + SAMT-247 group ( n = 4) and vaccine group ( n = 2). Data shown in b – i were analysed with the two-tailed Wilcoxon signed-rank test. Horizontal and vertical bars denote mean and standard deviation, respectively.

Article Snippet: CD14 + cells were isolated from cryopreserved PBMCs (10 × 10 6 cells) collected following pre-study and 2 weeks post last immunization (week 14) by using non-human primate CD14 MicroBeads (#130-091-097, Miltenyi Biotec) following manufacturer instructions.

Techniques: Imaging, Fluorescence, Expressing, Software, Comparison, Marker, Two Tailed Test, Standard Deviation

Journal: Nature Microbiology

Article Title: Vaccine plus microbicide effective in preventing vaginal SIV transmission in macaques

doi: 10.1038/s41564-023-01353-7

Figure Lengend Snippet: Vaccination-induced ADCC results in apoptosis of SIV-infected cells, which in turn are cleared by efferocytes to avoid inflammation and preserve tissue homeostasis. Vaccine-induced IL-10 expression in CD14 + monocytes further augments efferocytosis. Vaccine-induced NKp44 + cells produce the IL-17 cytokine that maintains mucosal epithelium integrity. All of these protective effector responses were enhanced dramatically in the vaccine + SAMT-247 group, increasing protection from SIV mac251 acquisition. The scheme is adapted from Bissa et al. .

Article Snippet: CD14 + cells were isolated from cryopreserved PBMCs (10 × 10 6 cells) collected following pre-study and 2 weeks post last immunization (week 14) by using non-human primate CD14 MicroBeads (#130-091-097, Miltenyi Biotec) following manufacturer instructions.

Techniques: Infection, Expressing

Journal: bioRxiv

Article Title: UBA1 Mutations Drive RIPK1-Mediated Cell Death and Monocyte Dysfunction in VEXAS Syndrome

doi: 10.1101/2025.10.06.680650

Figure Lengend Snippet: Regulated cell death pathways are activated in myeloid cells from VEXAS patients. ( A ) Leucocytes, neutrophils, monocytes, lymphocytes count from patients with VEXAS and elderly gender-matched healthy controls (HC). Panels show individual data (dots) and means ± SEM (histograms). P values were determined by the Mann-Whitney test. ( B ) Hematoxylin-eosin staining of UBA1-mutated Sweet-like lesion revealing karyorrhectic nuclei and apoptotic debris in VEXAS (arrows). Illustrative picture is shown (Magnification x10 and x40, scale bar 100 μm and 50µm). ( C ) Representative immunofluorescence images of pMLKL (green) staining on CD14+ sorted cells form 3 active VEXAS patients and 3 elderly gender-matched HC. Nuclei are stained in blue with Hoechst. Percentage of pMLKL cell surface area per CD14+ cells, data are shown and means (Histograms) ± SEM. Forty cells were quantified for each patient. ( D ) Multiplex Immunofluorescence of skin biopsy samples from VEXAS skin lesion showing the co-expression of cleaved GSDMD, MLKL, cleaved caspase-3 and phosphorylated RIPK1 within CD68+ infiltrates in VEXAS. ( E ) Gene set enrichment analysis of apoptosis (KEGG), necroptosis (GOBP) and pyroptosis (GOBP) pathways enriched in VEXAS lesional skin (n=6) versus non lesional skin from HC (n=5) adapted form from dataset GSE245639. *P□<□0.05; **P□<□0.01; ***P□<□0.001, ****P□<□0.0001. SEM, HC, Healthy Control; Standard Error of the Mean; VEXAS, Vacuoles, E1 enzyme, X-linked, Autoinflammatory, Somatic; WT, Wild-Type.

Article Snippet: For CD14+ cells form patients, cells were sorted from 2mL fresh Whole Blood collected in EDTA tube and sorted using StraightFrom® Whole Blood CD14 MicroBeads and Whole Blood column kit (Milteny Biotec, #130-090-879) according to the manufacturer’s instructions.

Techniques: MANN-WHITNEY, Staining, Immunofluorescence, Multiplex Assay, Expressing, Control

Journal: PLOS ONE

Article Title: CCX559 is a potent, orally-administered small molecule PD-L1 inhibitor that induces anti-tumor immunity

doi: 10.1371/journal.pone.0286724

Figure Lengend Snippet: (A) TNF secretion from five PBMCs donors was measured after 48 hours of treatment with CCX559 or DMSO, alone or in the presence of anti-CD3/CD28/CD2 microbeads. All donor PBMCs secreted TNF when stimulated by the CD3/CD28/CD2 beads, but CCX559 treatment alone induced no TNF secretion. (B) Soluble PD-L1 levels in cynomolgus monkey plasma were measured after 28 days of treatment with a vehicle control, 25 or 60 mg/kg CCX559. The change in sPD-L1 plasma levels was calculated by subtracting the concentration at predose from day 28 for each animal, and was significantly increased by 60 mg/kg CCX559 in both genders. The average predose plasma sPD-L1 levels ranged from 77 to 148 pg/ml and were not significantly different between groups. One-way ANOVA with Dunnett’s multiple comparisons was used to compare the day 28 change in sPD-L1 between dose groups for each gender. All data are represented as mean ± SD, *denotes p < 0.05, and ** p < 0.01.

Article Snippet: Monocytes isolated with human CD14 + MicroBeads (Miltenyi, 130-045-201) were differentiated to dendritic cells (moDCs) with GM-CSF (100 ng/ml) and IL-4 (50 ng/ml) for 6 days.

Techniques: Clinical Proteomics, Control, Concentration Assay