Journal: iScience

Article Title: Hypoxic-immune axis orchestrates metastatic dissemination via HIF isoform imbalance in pancreatic neuroendocrine tumors

doi: 10.1016/j.isci.2025.114340

Figure Lengend Snippet: Immune cell infiltration in KRAS G12C mutant PNETs (A) Quantitative analysis using Image Plus 6.0 software reveals a significant increase in Tregs (CD4 + , CD25 + , and FoxP3 + ) within KRAS- G12C mutant PNETs compared to wild-type KRAS PNET tissues, suggesting KRAS G12C -driven immunosuppressive cell recruitment. (B) Fluorescence intensity analysis demonstrates reduced infiltration of CD8 + cytotoxic T cells and HLA-DR + activated cells in KRAS G12C mutant tumors, indicative of impaired antitumor immunity. (C) Elevated MDSCs counts in KRAS G12C mutant PNETs correlate with enhanced immune evasion.

Article Snippet: APC Anti-Human CD8a Antibody [OKT-8] , Elabscience , E-AB-F1110E.

Techniques: Mutagenesis, Software, Fluorescence

Journal: iScience

Article Title: Hypoxic-immune axis orchestrates metastatic dissemination via HIF isoform imbalance in pancreatic neuroendocrine tumors

doi: 10.1016/j.isci.2025.114340

Figure Lengend Snippet: Tregs, CD8 + T cells, and HLA-DR + cells in KRAS G12C -mutated PNETs (A) Flow cytometry plots and fluorescence intensity histograms demonstrate elevated CD4 + T cell proportions in KRAS G12C patient blood samples compared to wild-type KRAS tumors and healthy controls. (B) Quantification shows a significant enrichment of CD25 + T cells in KRAS G12C patients, surpassing both wild-type KRAS tumors and normal controls. (C) Quantitative data and histogram overlays confirm a substantial increase in FoxP3 + T cells frequency in KRAS G12C patients, with levels moderately elevated compared to wild-type KRAS and significantly higher than healthy individuals. (D) A slight decrease in CD8 + T cell frequency in KRAS G12C samples relative to wild-type KRAS, with levels significantly lower than those in healthy individuals (E) A moderate reduction in HLA-DR + cell frequency in KRAS G12C patients compared to wild-type KRAS, alongside a marked suppression relative to healthy controls.

Article Snippet: APC Anti-Human CD8a Antibody [OKT-8] , Elabscience , E-AB-F1110E.

Techniques: Flow Cytometry, Fluorescence

Journal: Nature Communications

Article Title: Autologous HIV-specific T cell therapy targeting conserved epitopes is well-tolerated in six adults with HIV: an open-label, single-arm phase 1 study

doi: 10.1038/s41467-025-59810-2

Figure Lengend Snippet: a HST-NEET products ( n = 6) exhibited variable antigen-specific IFN-γ responses against HIV Gag, Pol, Nef, and conserved epitope targets in the Gag/Pol Mos1 and Mos2 peptide mixes. RESIST-004 HST-NEET numbers after treatment-related studies were insufficient to test Gag/Pol Mos1 and Mos2 specificity. Positive ELISpot results were defined as IFN-γ spot forming cells ≥2 times the actin negative control. Spot forming cells (SFC) per 10 5 cells indicative of specific responses compared to actin stimulated cells: RESIST-001 (actin [17 SFC], nef [139 SFC]), RESIST-003 (actin [19 SFC], gag [659 SFC], pol [466 SFC], nef [1057 SFC], mos1 [540 SFC], mos2 [520 SFC]), RESIST-004 (actin [34 SFC], nef [117 SFC]), RESIST-005 (actin [65 SFC], nef [804 SFC]), RESIST-006 (actin [0 SFC], nef [236 SFC]), and RESIST-007 (actin [25 SFC], gag [1063 SFC], nef [854 SFC], mos1 [291 SFC], mos2 [290 SFC]). b Each bar reports the total sum (infusion 1 and infusion 2) of infused T cells specific for HIV Gag, Pol, and Nef epitopes for each HST-NEETs product ( n = 5). c Each bar reports the total sum (infusion 1 and infusion 2) of infused T cells specific for conserved epitopes in Gag/Pol Mosaic 1 and Mosaic 2 immunogens for each HST-NEETs product ( n = 5). d , e HST-NEET products ( n = 6) were predominantly CD8+ effector memory T cells and had variable expression of T cell surface activation and exhaustion markers. f CD3+ CD8+ HST-NEET products ( n = 5) exhibited polyfunctional T cell responses upon stimulation with Gag, Pol, Nef and Gag/Pol Mos1 and Mos2 peptides as measured by intracellular production of IFN-γ and TNF-α. HIV-specific T cell activation was dominant in the CD8+ T cell fraction of all HST-NEET products. Source data are provided as a Source Data file.

Article Snippet: Antibodies used in panels for cell surface staining differed between the GMP and non-GMP laboratories, and were combinations of the following: CD3 PE-Cy7 (Cat: 317334, Biolegend), CD3 PerCP-Vio700 (Cat: 130-113-141, Miltenyi), CD3 BV785 (Cat: 317330, Biolegend), CD4 BV605 (Cat: 317438, Biolegend), CD4 PE Vio770 (Cat: 130-113-227, Miltenyi), CD8a BV421 (Cat: 301036, Biolegend), CD8a APC Vio770 (Cat: 130-110-681, Miltenyi), CCR7 Alexa Fluor 700 (Cat: 353244, Biolegend), CCR7 FITC (Cat: 130-120-468, Miltenyi), CD45RO PE-Dazzle 594 (Cat: 304248, Biolegend), CD45RO PE (Cat: 130-113-559, Miltenyi), CD19 FITC (Cat: 130-113-645, Miltenyi), CD14 VioBlue (Cat: 130-110-524, Miltenyi), CD16 PE (Cat: 130-113-393, Miltenyi), CD56 BV650 (Cat: 318344, Biolegend), CD56 PE (Cat: 130-113-312, Miltenyi), TCRγδ APC-Fire 750 (Cat: 331228, Biolegend), TCRγδ (Cat: 130-113-512, Miltenyi), TCRαβ PerCP/Cyanine5.5 (Cat: 306724, Biolegend), TCRαβ FITC (Cat: 130-113-538, Miltenyi), CD160 PE-Cy7 (Cat: 341212, Biolegend), CTLA4 PE-Dazzle 594 (Cat: 369616, Biolegend), LAG3 PE (Cat: 369306, Biolegend), PD-1 FITC (Cat: 329904, Biolegend), TIM3 BV650 (Cat: 345028, Biolegend), CD107a PE-Cy7 (Cat: 328618, Biolegend), CD45 APC (Cat: 130-110-633, Miltenyi), CD83 PE (Cat: 130-110-503, Miltenyi), CD62 VioBlue (Cat: 130-113-622, Miltenyi), HLA-DR FITC (Cat: 30-111-788, Miltenyi), and CD95 APC (Cat: 558814, BD Biosciences).

Techniques: Enzyme-linked Immunospot, Negative Control, Expressing, Activation Assay

Journal: Nature Communications

Article Title: Autologous HIV-specific T cell therapy targeting conserved epitopes is well-tolerated in six adults with HIV: an open-label, single-arm phase 1 study

doi: 10.1038/s41467-025-59810-2

Figure Lengend Snippet: a Comprehensive pooled peptide epitope mapping of HST-NEETs revealed 15-mer peptides driving HIV-specific responses in each product ( n = 5). RESIST-001, -003, -005, and -006 ELISpots were conducted as singlets based on HST-NEET numbers available for characterization studies, and RESIST-007 ELISpots were conducted in duplicate. Plotted SFC per 10 5 cells is shown above each bar. b – f HST-NEET products were stimulated with 15-mer peptides in ( a ) for 5 h and intracellular production of IFN-γ and TNF-α in CD8+ T cells was evaluated to confirm epitope-specific responses. Peptides evaluated are named by the first two amino acids in the 15-mer sequence followed by the peptide length. g Minimal (9 amino acid) epitopes associated with CD8+ T cell responses were determined for each product. RESIST-004 HST-NEETs were unavailable for these product characterization studies. Positive ELISpot results were defined as IFN-γ spot forming cells ≥2 times the actin negative control. RESIST-005 ELISpots were conducted as singlets, RESIST-001, -003, -006, and -007 were conducted in duplicate. Plotted SFC per 10 5 cells is shown above each bar.

Article Snippet: Antibodies used in panels for cell surface staining differed between the GMP and non-GMP laboratories, and were combinations of the following: CD3 PE-Cy7 (Cat: 317334, Biolegend), CD3 PerCP-Vio700 (Cat: 130-113-141, Miltenyi), CD3 BV785 (Cat: 317330, Biolegend), CD4 BV605 (Cat: 317438, Biolegend), CD4 PE Vio770 (Cat: 130-113-227, Miltenyi), CD8a BV421 (Cat: 301036, Biolegend), CD8a APC Vio770 (Cat: 130-110-681, Miltenyi), CCR7 Alexa Fluor 700 (Cat: 353244, Biolegend), CCR7 FITC (Cat: 130-120-468, Miltenyi), CD45RO PE-Dazzle 594 (Cat: 304248, Biolegend), CD45RO PE (Cat: 130-113-559, Miltenyi), CD19 FITC (Cat: 130-113-645, Miltenyi), CD14 VioBlue (Cat: 130-110-524, Miltenyi), CD16 PE (Cat: 130-113-393, Miltenyi), CD56 BV650 (Cat: 318344, Biolegend), CD56 PE (Cat: 130-113-312, Miltenyi), TCRγδ APC-Fire 750 (Cat: 331228, Biolegend), TCRγδ (Cat: 130-113-512, Miltenyi), TCRαβ PerCP/Cyanine5.5 (Cat: 306724, Biolegend), TCRαβ FITC (Cat: 130-113-538, Miltenyi), CD160 PE-Cy7 (Cat: 341212, Biolegend), CTLA4 PE-Dazzle 594 (Cat: 369616, Biolegend), LAG3 PE (Cat: 369306, Biolegend), PD-1 FITC (Cat: 329904, Biolegend), TIM3 BV650 (Cat: 345028, Biolegend), CD107a PE-Cy7 (Cat: 328618, Biolegend), CD45 APC (Cat: 130-110-633, Miltenyi), CD83 PE (Cat: 130-110-503, Miltenyi), CD62 VioBlue (Cat: 130-113-622, Miltenyi), HLA-DR FITC (Cat: 30-111-788, Miltenyi), and CD95 APC (Cat: 558814, BD Biosciences).

Techniques: Sequencing, Enzyme-linked Immunospot, Negative Control