cd4 cd8α cd19 cd56 cells  (Miltenyi Biotec)

Journal: Allergy, Asthma, and Clinical Immunology : Official Journal of the Canadian Society of Allergy and Clinical Immunology

Article Title: Effects of sublingual immunotherapy for dust mite on Th 17 / Treg cells in children with asthma

doi: 10.1186/s13223-026-01010-8

Figure Lengend Snippet: Increases in Treg cells following treatment. FACS was used to detect CD4+CD25+Foxp3+IL-10+ T cells in patients of SLIT group and non-SLIT group. Treg cell percentage significantly increased throughout the research period in SLIT group (F = 40.267, p < 0.01); Treg cell were unchanged throughout the study period in non-SLIT group (F = 3.525, p = 0.172). ** p < 0.01

Article Snippet: The fluorescein-isothiocyanate (FITC) mouse anti-human CD4, mouse anti-human (PE Alexa Fluor 700), Alexa Fluor 647 anti-human IL-10, APC anti-human IL-17, PE anti-human Foxp3, anti-human CD25, human IL-6, human IL-10, human IL-17 ELISA kits were all purchased from eBioscience, Inc (San Diego, CA, USA).

Techniques:

Journal: bioRxiv

Article Title: NK cell immunotherapy administered at the time of HIV recrudescence is associated with viral control

doi: 10.1101/2025.11.19.688897

Figure Lengend Snippet: A) Schematic figure showing the overview of NK cell production and the overview of the mouse study timeline. Red circles on the timeline indicate blood collection, black dots indicate scheduled necropsies. Schematic of transgene design: MND promoter, CAR (CD4-MBL-CD8TM-4-1BB-CD3z), CXCR5, IL-15, the BgH PolyA tail, with separation mediated by P2A and T2A sites, and inverted terminal repeats (ITRs). Schematic showing expression of all transgene components as well as base editor-mediated knock out of PD-1 on the NK cell surface. Figure made using Biorender.com. B) Expression of CAR (MBL) and CXCR5 molecules was confirmed in the cell product using flow cytometry. Cells were pre-gated sequentially on lymphocytes, singlets, live cells, CD56+, and CD3-. C) PD-1 knockout was detected at the RNA level by RT-PCR, CAR NK (blue) relative to control NK cells (black). D) IL-15 expression in CAR NK (blue) and control NK (black) cells via IL-15 ELISA of cell culture supernatant. E) CAR (blue) and control (black) cell proliferation over one week post-thaw. F) The specific lysis of HIV Envelope P815s by CAR (blue) or control (black) NK cells via DELFIA cytotoxicity assay. 5:1, 10:1, and 20:1 E:T ratios were tested. All assays were performed in duplicate or triplicate. Data are represented as mean + standard deviation (error bars).

Article Snippet: After a 2-day recovery, CAR NK cells were sorted on CD56+ CD4+ using the MACSQuant Tyto Cell Sorter (Miltenyi Biotec) and expanded for an additional 7 days following the procedure described above.

Techniques: Expressing, Knock-Out, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay, Cell Culture, Lysis, Cytotoxicity Assay, Standard Deviation

Journal: bioRxiv

Article Title: NK cell immunotherapy administered at the time of HIV recrudescence is associated with viral control

doi: 10.1101/2025.11.19.688897

Figure Lengend Snippet: NK (CD56+) cells in CAR (blue), control (black), or PBS (red) treated groups from A) Peripheral Blood, B) Lymph node (at necropsies), and C) Spleen (at necropsies). CAR+CXCR5+ NK cells over time in D) Peripheral Blood, E) Lymph node (at necropsies), and F) Spleen (at necropsies). Flow was pre-gated on Lymphocytes, Singlets, Live Cells, human CD45+, mouse CD45-, CD3+, and then through CD56+ MBL+ CXCR5+ (see Fig. S4 for gating strategy). Median viral loads (green), NK cell levels (orange), post-treatment in G) controllers or H) non-controllers. I) Median human hematopoietic cell counts (CD45+)/ ul blood of controllers (gray) or non-controllers (pink).

Article Snippet: After a 2-day recovery, CAR NK cells were sorted on CD56+ CD4+ using the MACSQuant Tyto Cell Sorter (Miltenyi Biotec) and expanded for an additional 7 days following the procedure described above.

Techniques: Control

Journal: bioRxiv

Article Title: NK cell immunotherapy administered at the time of HIV recrudescence is associated with viral control

doi: 10.1101/2025.11.19.688897

Figure Lengend Snippet: CD4:CD8 ratios over time in A) CAR NK-treated animals (blue), B) control NK-treated animals (black), and C) PBS-treated animals (red). CD4:CD8 ratios in CAR (blue), control NK (black), and PBS (red) treated groups at D) -6 DPT, E) 6 DPT, F) 14 DPT, G) 28 DPT, H) 42 DPT, and I) 56 DPT. Lines represent median values. CD4:CD8 ratios were determined by flow cytometry and pre-gated on Lymphocytes, Singlets, Live Cells, Human CD45+, Mouse CD45-, CD3+ (see gating strategy in Fig. S4 ).

Article Snippet: After a 2-day recovery, CAR NK cells were sorted on CD56+ CD4+ using the MACSQuant Tyto Cell Sorter (Miltenyi Biotec) and expanded for an additional 7 days following the procedure described above.

Techniques: Control, Flow Cytometry

Journal: Precision Clinical Medicine

Article Title: Single-cell transcriptomic profiling reveals distinct tumor microenvironments in HPV-associated penile squamous cell carcinoma

doi: 10.1093/pcmedi/pbaf013

Figure Lengend Snippet: CD8 + T cells display a lower expression of ICB checkpoints in HPV + PSCC. ( A – C ) UMAP plot of 14 693 TIL-Ts in Fig. . Each dot represents one single cell, coloured according to 11 clusters ( A ), 11 patients ( B ), and HPV status ( C ). These clusters were grouped into five cell types, including CD4 + T, PreT, CD8 + T, individual T (PT6), and proliferation T cells. Individual T cells were excluded for downstream statistical proportion analysis to reduce analytical bias. ( D ) UMAP visualization showing the classical marker gene expression to define major cell types. ( E ) Relative proportion of each cluster in 10 PSCC patients from HPV − and HPV + PSCC. ( F ) Multiplex immunofluorescence depicting the distribution of CD4 + and CD8 + T cells in HPV + and HPV − PSCC. Representative images include DAPI + nuclei for reference. Scale bar: 200 μm. ( G ) Quantification of CD4 + and CD8 + T cells per area between HPV + and HPV − PSCC ( n = 3). GSEA showing pathways significantly enriched in HPV − PSCC ( H, I ) or HPV + PSCC ( J, K ). Violin plot showing the expression of cytokine ( L ) or immune checkpoint ( M ) in CD8 + T cells by HPV status. Statistical analysis was performed using a two-sided unpaired Student's t -test for (G) and a Wilcoxon rank sum test for (L) and (M). Data are presented as mean ± SEM, with significance levels indicated as follows: ns: not significant, * P < 0.05. ICB: Immune checkpoint blockade.

Article Snippet: For IF staining, FFPE tissue slices were treated the same as for IHC staining, then incubated with mixed primary antibodies overnight (mouse anti-human CD4 antibody ZsBio ZM-0418; mouse anti-human CD8 antibody, Servicebio GB12068; HPV E7 antibody, Abcam ab308296; HPV E6 antibody Santa cruz sc460), incubated with fluorescein-conjugated secondary antibodies, and finally counterstained with DNA dye DAPI (4′,6-diamidino-2-phenylindole), mounted with anti-fading mounting medium, and stored at 4 °C in the dark until imaging.

Techniques: Expressing, Marker, Gene Expression, Multiplex Assay, Immunofluorescence