selection human cd14 microbead kit  (Miltenyi Biotec)

Journal: bioRxiv

Article Title: IFI16/AIM2 inflammasomes control Gal-9 and PVR in myeloid cells from PWH and their targeting improves immunotherapy against HIV-1

doi: 10.64898/2026.01.26.701473

Figure Lengend Snippet: (A): ELISA analysis of plasma concentration of IL-1β, comparing PWH with normal CD4/CD8 T cell ratio (defined as ≥1) and low CD4/CD8 T cell ratio (defined as < 0.8). Statistical significance was calculated using two-tailed Wilcoxon test. (B): Spearman correlation between plasma concentrations of soluble CD14 (ssCD14) and IL-1β. P and R values are shown (C): Expression of Gal-9 (Gal9; higher plot) or PVRhi (lower plot) in activated CD40hi in Mo from HIV- 1-negative controls, and PWH with low CD4/CD8 T cell ratio or normal CD4/CD8 T cell ratio after 16-hour stimulation with PAMPs mimicking bacterial translocation (LPS, red; Flagellin, Flag, purple) or viral replication (CL097, ssRNA, orange; Poly I:C, dsRNA, blue). Statistical significance was calculated using Kruskal-Wallis test for comparison of stimulation with the same PAMP between groups (*p<0.05; **p<0.01), and One-Way ANOVA Friedman test with Dunn’s multiple comparison test for comparison within the same group (*p<0.05; **p<0.01; ***p<0.001; ****p<0.0001).

Article Snippet: Mo were isolated from PBMC from our study cohort by the human CD14 MicroBeads kit positive immunomagnetic selection using MS Columns (Miltenyi Biotec).

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Concentration Assay, Two Tailed Test, Expressing, Translocation Assay, Comparison

Journal: bioRxiv

Article Title: IFI16/AIM2 inflammasomes control Gal-9 and PVR in myeloid cells from PWH and their targeting improves immunotherapy against HIV-1

doi: 10.64898/2026.01.26.701473

Figure Lengend Snippet: (A-B): Histological analysis of IFI-16 (A) or AIM2 (B) (green) and Caspase-1 (red) expression and DAPI (blue) in the spleen from a viremic HIV-1 infected humanized BLT mouse. Quantifications of cells positive cells for each inflammasome sensor alone (top plots), co-expressing Caspase 1 (CASP1) (middle plots) or both Caspase 1 and CD14 (bottom plots) are shown on the right. (C): Histological analysis of Gal-9 (Gal9; green), CD14 (white) and Caspase-1 (Casp1, red) expression and DAPI (blue) in the spleen from a viremic HIV-1 infected humanized BLT mouse. Zoomed area demonstrating co-expression between Gal9, CD14 and Caspase-1 are shown on the right. Arrows highlight cells co-expressing the mentioned markers

Article Snippet: Mo were isolated from PBMC from our study cohort by the human CD14 MicroBeads kit positive immunomagnetic selection using MS Columns (Miltenyi Biotec).

Techniques: Expressing, Infection

Journal: bioRxiv

Article Title: Trogocytosis-mediated transfer of FOLR2 from Nurse-like cells to CLL cells is linked to their activation and proliferation

doi: 10.1101/2024.12.31.630890

Figure Lengend Snippet: A) FC analysis of the expression of FOLR2, CD14, CD163 and CD206 on cells from a culture of PBMC from a CLL patient at day 12 of in vitro culture, results for big, granular cells corresponding to NLCs (upper panel) and small cells corresponding to lymphoid cells (bottom panel). B) Expression of FOLR2, CD14, CD163, CD206 on the small cells (lymphocytes including CLL cells) of CLL PBMC cultures at day 12 (19 donors). C) FOLR2 expression on CLL cells (CD5 + CD19 + ) alone at day 0 or in PBMC culture on day 12. D) left: FOLR2 expression by CLL cells in co-culture with CTRL HD-NLCs or FOLR2 KO NLCs; right: Percentage of FOLR2 + CLL cells in co-cultures with CTRL HD-NLCs or FOLR2 KO HD-NLCs (10 independent experiments).

Article Snippet: Healthy donor monocytes – following a buffy coat processing (EFS - Établissement français du sang, France), PBMC were subjected to CD14 MicroBeads positive selection kit with use of MS/LS columns (Miltenyi Biotec, France), according to the manufacturer’s protocol.

Techniques: Expressing, In Vitro, Co-Culture Assay

Journal: bioRxiv

Article Title: Trogocytosis-mediated transfer of FOLR2 from Nurse-like cells to CLL cells is linked to their activation and proliferation

doi: 10.1101/2024.12.31.630890

Figure Lengend Snippet: A) FC analysis of expression of Ki67, CD23, CD184 and FOLR2 on CLL cells cultured alone or with HD-NLCs in the presence of CD40L+Il15. B) Percentage of Ki67 + CLL cells cultured alone or with HD-NLCs in the presence of CD40L+IL-15. C) Correlation between FOLR2 acquisition and Ki67 expression by CLL cells co-cultured with HD-NLCs in the presence of CD40L+IL-15 (5 independent experiments, 12 patients, >200 samples). D) CTV dilution assay of CLL cells cultured alone or with HD-NLCs in the presence of CD40L+IL-15: exemplary FC results (left), cumulative results from 8 patients represented as percentage of proliferating cells based on decrease in CTV signal (right). E) Percentage of actively cycling (Ki67 + , left) and total fraction of cycling (CTV low, right) CLL cells in co-culture with HD-NLCs in the presence of CD40L+IL-15, in the media with various concentrations of folic acid (FA). F) FC analysis of CD23, CD184, CD14 and FOLR2 signal on CLL cells stratified based on Ki67/CTV level (P1. CTV high Ki67 - : quiescent cells that did not divide; P2. CTV high Ki67 + : cells right before the first division; P3. CTV low Ki67 + : actively cycling cells after previous division; P4. CTV low Ki67 - : cells after cycling, quiescent again). G) FC analysis of FOLR2 level represented as MFI on CLL cells, depending on the proliferative status (Ki67/CTV) and subsequent cell generations (G1-G5).

Article Snippet: Healthy donor monocytes – following a buffy coat processing (EFS - Établissement français du sang, France), PBMC were subjected to CD14 MicroBeads positive selection kit with use of MS/LS columns (Miltenyi Biotec, France), according to the manufacturer’s protocol.

Techniques: Expressing, Cell Culture, Dilution Assay, Co-Culture Assay

Journal: bioRxiv

Article Title: Trogocytosis-mediated transfer of FOLR2 from Nurse-like cells to CLL cells is linked to their activation and proliferation

doi: 10.1101/2024.12.31.630890

Figure Lengend Snippet: A) FC analysis of the expression of FOLR2 on a subpopulation of NLCs (CD163 + CD206 + ) generated in a culture of PBMC from a CLL patient (grey: isotypic control; green: anti-FOLR2). B) Comparison of FOLR2 expression on various types of in vitro generated macrophages: NLCs, M2-like and M1-like macrophages (Left: representative histograms; right: compilation of 9 independent experiments. C) Kinetics of FOLR2 expression during CD14 + monocyte to NLC differentiation in CLL PBMC cultures for 14 days measured by FC (MFI ratio: anti-FOLR2/isotopic control). D) FC analysis of the CLL cells viability from CLL PBMC cultures on day 12 (annexin V/7-AAD staining). E) Percentage of CLL cells viability for the three groups of CLL patients according to the FOLR2 expression on NLCs. F) Correlation of the FOLR2 expression on NLCs with the CLL cells viability. G) Multiplex immunofluorescence (mIF) of a reactive lymph node and a lymph node from a CLL patient with the staining of CD20 (cyan), FOLR2 (yellow) and CD163 (purple). H) Representative images of lymph nodes from CLL patients, with various stages of the disease, from indolent to accelerated and Richter transformation; stained with anti-CD20 (cyan), anti-FOLR2 (yellow) and anti-CD163 (purple) antibodies (objective: x20).

Article Snippet: Healthy donor monocytes – following a buffy coat processing (EFS - Établissement français du sang, France), PBMC were subjected to CD14 MicroBeads positive selection kit with use of MS/LS columns (Miltenyi Biotec, France), according to the manufacturer’s protocol.

Techniques: Expressing, Generated, Control, Comparison, In Vitro, Staining, Multiplex Assay, Immunofluorescence, Transformation Assay

Journal: bioRxiv

Article Title: Trogocytosis-mediated transfer of FOLR2 from Nurse-like cells to CLL cells is linked to their activation and proliferation

doi: 10.1101/2024.12.31.630890

Figure Lengend Snippet: A) FC analysis of the PKH67/FOLR2/CD14/CD16 signal on CLL cells after 4-hours of co-culture with PKH67 + NLCs (upper: CLL cells alone; below: co-culture with NLCs). B) Percentage of CLL cells positive for PKH67, FOLR2, CD14 and CD16 after 4-hours of co-culture with NLCs (15 independent experiments). C-E) Confocal microscopy (x63 magnification) of co-culture of CLL cells (CD19 + : cyan) and NLCs (FOLR2 + : red) during 4-hours showing: the contact of NLCs and CLL cells ( C ), the acquisition of FOLR2 patches by CLL cells after contact with NLCs ( D ) and the absence of detection of FOLR2 signal on CLL cells after co-culture with FOLR2 KO HD-NLCs ( E ). F) Confocal microscopy of a 4-hours co-culture of CLL cells with CTRL HD-NLCs (upper) or with FOLR2 KO NLCs (below) incubated with folic acid-FITC (green) then stained with anti-FOLR2-CF633 (red) and with CD19-BV421 (cyan). G) Confocal microscopy of purified CLL cells after co-culture with CTRL HD-NLCs (upper) or FOLR2 KO HD-NLCs (below), following by incubation with folic acid-FITC for 90 min.

Article Snippet: Healthy donor monocytes – following a buffy coat processing (EFS - Établissement français du sang, France), PBMC were subjected to CD14 MicroBeads positive selection kit with use of MS/LS columns (Miltenyi Biotec, France), according to the manufacturer’s protocol.

Techniques: Co-Culture Assay, Confocal Microscopy, Incubation, Staining, Purification

Journal: JCI Insight

Article Title: The histone methyltransferase Mixed-lineage-leukemia-1 drives T cell phenotype via Notch signaling in diabetic tissue repair

doi: 10.1172/jci.insight.179012

Figure Lengend Snippet: ( A ) River plots depicting cell groups and ligand-receptor pairs affect outgoing (signal source) and incoming (signal responder) pattern. The thickness of the flow indicates the contribution of the cell group or signaling pathway to each latent pattern ( n = 10). ( B ) Human bulk RNA-Seq heatmap reflecting the expression profiles for selective genes (rows) across different samples (columns; stratified by T2D status) from Gene Ontology pathway analysis with upregulation of Notch signaling in T2D wounds compared with control wounds ( n = 7). ( C ) Notch receptor expression in wound CD4 + Th cells 7 days after wounding between diet-induced obesity (DIO) mice and their normal diet, nondiabetic littermate controls ( n = 6 mice/group, run in triplicate). ( D ) Analytical flow cytometry of wound cell suspensions 5 days after wounding between ND ( n = 4) and DIO ( n = 4) mice, evaluating for Lin – /CD3 + /CD4 + /Notch1 + (or Lin – /CD3 + /CD4 + /Notch2 + ). ( E ) Hes1 , Hey1 , and Hey2 expression in wound CD4 + Th cells 5 days after wounding between ND and DIO mice ( n = 6 mice/group, run in triplicate). ( F ) DLL4 expression in peripheral human CD14 + monocytes between nondiabetic ( n = 8) and diabetic ( n = 9) donors. ( G ) Analytical flow cytometry between ND ( n = 3) and DIO ( n = 3) wound Mφs evaluating for Lin − /Ly6G − /CD11b + /DLL4 + . * P < 0.05, *** P < 0.001, **** P < 0.0001. Data are presented as the mean ± SEM. Data were first analyzed for normal distribution, and if data passed the normality test, 2-tailed Student’s t test was used.

Article Snippet: Cell suspensions were then treated with anti–human CD14 microbeads (EasySep Human CD14 Positive Selection Kit, Stemcell Technologies) and purified by MACS as described above.

Techniques: RNA Sequencing, Expressing, Control, Flow Cytometry