Journal: Emerging Microbes & Infections

Article Title: Predictive markers of SARS-CoV-2 spike-specific cytotoxic T cell activity following Omicron breakthrough infection

doi: 10.1080/22221751.2025.2602317

Figure Lengend Snippet: Schematic overview of PBMC stimulation and experimental design. Serial PBMC samples were recovered overnight and a fraction applied for 24 h to ELISpot plates with either vehicle (negative control), the full SARS-CoV-2 S peptide pool, anti-CD3 (positive control) or individual peptides to assess circulating S-specific T cell frequency by IFN-γ and/or IL-2 production. Remaining PBMC were stimulated with the full SARS-CoV-2 S peptide pool or individual peptides in the presence of IL-7 with IL-2 added on day 3. On day 7, an aliquot of the cultured cells was restimulated for 5 h with either vehicle, the full SARS-CoV-2 S peptide pool or individual peptides, followed by flow cytometric analysis of individual T cell subsets for IFN-γ and IL-2 production. On day 10, the remaining expanded cells were assessed for CTL activity using a 51 Cr-release assay against autologous BLCL pulsed with vehicle, the full SARS-CoV-2 S peptide pool, or individual peptides selected based on donor HLA typing. A matrix approach was used to deconvolute unknown CTL epitopes using BLCLs pulsed with matrix peptide pools, with confirmation using individual peptides from BEI on days 11-12.

Article Snippet: Effector T cells were stained with directly conjugated mAb against human CD3 (VioGreen, REA613, Miltenyi Biotec), CD4 (APC-Vio770, REA623, Miltenyi Biotec), CD8 (AlexaFluor® 700, HIT8a, BioLegend), CD57 (FITC, NK-1, BD Biosciences, Toronto, ON, Canada), and IFN-γ (APC, 4S.B3, Invitrogen) and IL-2 (PE, MQ1-17H12, Invitrogen) using the Inside Stain Kit (Miltenyi Biotec) as per manufacturer’s instructions and as previously described in [ ].

Techniques: Enzyme-linked Immunospot, Negative Control, Positive Control, Cell Culture, Activity Assay, Release Assay, Immunopeptidomics

Journal: Emerging Microbes & Infections

Article Title: Predictive markers of SARS-CoV-2 spike-specific cytotoxic T cell activity following Omicron breakthrough infection

doi: 10.1080/22221751.2025.2602317

Figure Lengend Snippet: Circulating S-specific SARS-CoV-2 T cell activity. Circulating T cell responses were measured over 24 h in ELISpot assays (in duplicate) using vehicle control, full SARS-CoV-2 S peptide pool, anti-CD3 or individual peptides. Persons with two (PV2; blue, right panel; n = 6) or three (PV3; pink, left panel; n = 22) vaccinations followed by Omicron breakthrough infection (POMI) were tested for (A) IFN-γ, (B) dual IFN-γ/IL-2, and (C) IL-2 SFC/10 6 PBMC. Data are shown as median with IQR (PV3/POMI) and as mean ± SD (PV2/POMI). Groups compared are spanned by lines above them with significant differences ( p value) or non significance (ns) indicated on the line. Wilcoxon matched-pairs signed rank test or Student’s paired t -test was used as appropriate for comparisons based on normality of data distribution.

Article Snippet: Effector T cells were stained with directly conjugated mAb against human CD3 (VioGreen, REA613, Miltenyi Biotec), CD4 (APC-Vio770, REA623, Miltenyi Biotec), CD8 (AlexaFluor® 700, HIT8a, BioLegend), CD57 (FITC, NK-1, BD Biosciences, Toronto, ON, Canada), and IFN-γ (APC, 4S.B3, Invitrogen) and IL-2 (PE, MQ1-17H12, Invitrogen) using the Inside Stain Kit (Miltenyi Biotec) as per manufacturer’s instructions and as previously described in [ ].

Techniques: Activity Assay, Enzyme-linked Immunospot, Control, Infection

Journal: Emerging Microbes & Infections

Article Title: Predictive markers of SARS-CoV-2 spike-specific cytotoxic T cell activity following Omicron breakthrough infection

doi: 10.1080/22221751.2025.2602317

Figure Lengend Snippet: IFN-γ and IL-2 production from expanded SARS-CoV-2 S-specific CD8 + T cells. (A) Representative gating strategy for identifying CD3 + CD8 + T cells. On day 7, expanded T cells were restimulated for 5 h with the full SARS-CoV-2 S peptide pool. Cytokine production was assessed for (B) IFN-γ, (C) dual IFN-γ/IL-2, and (D) IL-2 expression. Background signals from vehicle controls were subtracted before plotting. Data are shown as median with IQR (PV3/POMI n = 22; PV2/POMI n = 6). (E) Correlation between IL-2 production by CD8 + T cells on day 7 and IFN-γ spot-forming units per 10⁶ total T cells on day 1 was assessed. P values in (B-D) were calculated using Wilcoxon matched-pairs signed rank test and are shown above lines spanning the groups compared when significant or as ns when not significant. Significance of correlation in (E) was assessed using Spearman’s correlation.

Article Snippet: Effector T cells were stained with directly conjugated mAb against human CD3 (VioGreen, REA613, Miltenyi Biotec), CD4 (APC-Vio770, REA623, Miltenyi Biotec), CD8 (AlexaFluor® 700, HIT8a, BioLegend), CD57 (FITC, NK-1, BD Biosciences, Toronto, ON, Canada), and IFN-γ (APC, 4S.B3, Invitrogen) and IL-2 (PE, MQ1-17H12, Invitrogen) using the Inside Stain Kit (Miltenyi Biotec) as per manufacturer’s instructions and as previously described in [ ].

Techniques: Expressing

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet: Migratory DCs associate with tumor immune status, patient prognosis, and immunotherapeutic response in patients with cancer (A and B) Representative images of multiplex immunostaining of migratory DC and T cell markers on tumor sections derived from immune-“hot” and “cold” patients with PDAC/NSCLC are given. (C and D) Correlation analysis between migratory DC population and overall survival across our collected PDAC and NSCLC patient cohorts ( n = 66 patients with PDAC, n = 65 patients with NSCLC). (E and F) ROC curves represent the accuracy of migratory DCs and CD3 + CD8 + T cells in predicting overall survival in patients with PDAC and NSCLC. (G and H) Spatial analysis of multiplex IF images from each group. Magnified images are given. (I and J) Stack bar charts show the density of CD3 + CD8 + T cells at varying distances from migratory DCs within both hot and cold tumors from patients with PDAC/NSCLC. (K) Representative images of multiplex immunostaining of migratory DC and T cell markers on tumor sections from NSCLC immunotherapy non-responder and responder. (L) High population of migratory DCs correlated with better progression-free survival in patients with advanced NSCLC treated with anti-PD-1 ( n = 19 patients). (M) ROC curves illustrate the predictive accuracy of migratory DCs and CD3 + CD8 + T cells for progression-free survival in patients with advanced NSCLC treated with anti-PD-1. (N) Spatial proximity analysis of multiplex IF images from each group. Magnified image is given. (O) Stack bar chart illustrates the density of CD3 + CD8 + T cells at different distances from migratory DCs in each group. (C, D, and L) Log rank (Mantel-Cox). Scale bars in (A), (B), and (K) represent 50 μm. See also Figure S1 and Tables S1 , , and .

Article Snippet: rabbit mAb against mouse CD3 , CST , Cat# 78588S; RRID:AB_2889902.

Techniques: Multiplex Assay, Immunostaining, Derivative Assay

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet: cNP cancer cell @MV DC treatment triggers DC maturation and CD8 + T and NK cell activation in vitro (A and B) FACS analysis measured CD86 expression in immature human/mouse DCs after treatments. Bar charts display the percentage of CD11c + CD86 + DCs per group ( n = 3 independent experiments). (C and D) FACS analysis examined human/murine CD3 + CD8 + T cell proliferation after treatments ( n = 3 independent experiments). (E–H) FACS analysis assessed human/murine CD3 + CD8 + T cell activation after treatments. Bar charts show the percentage of CD3 + CD8 + CD69 + T cells or CD3 + CD8 + IFN-γ + T cells per group ( n = 3 independent experiments). (I–L) FACS analysis assessed human/murine NK cell activation after treatments. Graphs show the percentage of CD3 − CD56 + CD69 + , CD3 − NK1.1 + CD69 + , CD3 − CD56 + IFN-γ + , or CD3 − NK1.1 + IFN-γ + relative to the total human/murine NK population in each group ( n = 3 independent experiments). (M and N) Schematic diagrams illustrate LDH release assays conducted on PDAC patient-derived tumor organoids (T, target), using varying ratios of CD3 + CD8 + T cells (E, effector) pre-treated with the indicated treatments (M), or co-cultured with hDCs pre-treated with the indicated treatments (N). Bar charts indicate the killing efficiency percentage at each E:T ratio among different groups ( n = 3 independent experiments). (O and P) Cytotoxicity LDH release assays on DT6066 cells were conducted using different ratios of CD3 + CD8 + T cells that had been either pre-treated with the indicated treatments (O) or co-cultured with DCs pre-treated with the indicated treatments (P) ( n = 3 independent experiments). (A–L) One-way ANOVA. (M–P) Two-way ANOVA. See also Figure S3 .

Article Snippet: rabbit mAb against mouse CD3 , CST , Cat# 78588S; RRID:AB_2889902.

Techniques: Activation Assay, In Vitro, Expressing, Derivative Assay, Cell Culture

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet: cNP DT6066 @MV DC treatment converts immune-cold pancreatic tumors into hot tumors, reduces hypoxia, and enhances blood vessel function (A) FACS analysis of CD11c + CD86 + DCs in orthotopic DT6066 pancreatic tumors after treatments, with a bar chart showing their percentage relative to total lymphocytes ( n = 3 mice per group). (B) FACS analysis of CD3 + CD4 + CD69 + T cells in orthotopic pancreatic tumors after treatments, with a bar chart showing their percentage relative to total CD3 + CD4 + T cells in each group ( n = 3 mice per group). (C) FACS analysis of CD3 + CD8 + CD69 + T cells in orthotopic pancreatic tumors after treatments, with a bar chart showing their percentage relative to total CD3 + CD8 + T cells in each group ( n = 3 mice per group). (D) Representative images of CD11c and CD86 co-immunostaining in tumor sections from each treatment group. Bar chart shows the quantification of CD11c + CD86 + DC × 10 3 per cm 2 in each group ( n = 3 mice per group). (E and F) Representative images of co-immunostaining for CD3 and CD4 (E) or CD8 (F) in tumor sections from each treatment group ( n = 3 mice per group). (G) FACS analysis of CD3 − NK1.1 + NK cells or CD3 − NK1.1 + CD69 + NK cells in orthotopic pancreatic tumors after treatments. Bar charts show their percentage relative to total CD3 − cells (left) or CD3 − NK1.1 + NK cells (right) in each group ( n = 3 mice per group). (H–J) Cytotoxicity LDH release assays of DT6066 cells after co-culture with various ratios of CD3 + CD8 + T cells isolated from tumors (H), lymph nodes (I), or spleens (J) of orthotopic DT6066 pancreatic tumor-bearing mice after treatments ( n = 3 mice per group). (K) FACS analysis of migratory DCs in orthotopic pancreatic tumors and TDLNs from mice treated as indicated. Bar charts show CD86 + CD103 + DCs as a percentage of CD11c + DCs in each group ( n = 3 mice per group). (L) Representative images of CD11c/CD86/CD103 triple immunostaining on paired tumor and TDLN sections in each group. Bar chart shows the percentage of CD11c + CD86 + CD103 + migratory DC × 10 3 per cm 2 in tumors (left) or TDLNs (right) from each group ( n = 3 mice per group). (M–O) Representative microbubble contrast ultrasound images are shown, and bar charts show quantification across the entire tumors, including tumor cores ( n = 3 mice per group). (P) Representative IHC staining of endomucin in tumor sections from each treatment group. Bar chart shows blood vessel diameter (left) or number of blood vessels (right) per cm 2 in each group ( n = 3 mice per group). (Q) Representative image of GLUT1 and endomucin co-immunostaining in tumor sections from each group. Bar chart shows the relative GLUT1 intensity per group ( n = 3 mice per group). (A–G, K, L, and N–Q) One-way ANOVA. (H–J) Two-way ANOVA. Scale bars in (D), (E), (F), and (L) represent 50 μm. (M) 1 cm. (P and Q) 100 μm. See also Figures S5 and .

Article Snippet: rabbit mAb against mouse CD3 , CST , Cat# 78588S; RRID:AB_2889902.

Techniques: Immunostaining, Co-Culture Assay, Isolation, Triple Immunostaining, Immunohistochemistry

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet:

Article Snippet: rabbit mAb against mouse CD3 , CST , Cat# 78588S; RRID:AB_2889902.

Techniques: Virus, Recombinant, Negative Staining, Lysis, Control, Phospho-proteomics, Cell Isolation, DNA Purification, DNA Extraction, Bicinchoninic Acid Protein Assay, Protein Purification, Magnetic Beads, Software

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet: Migratory DCs associate with tumor immune status, patient prognosis, and immunotherapeutic response in patients with cancer (A and B) Representative images of multiplex immunostaining of migratory DC and T cell markers on tumor sections derived from immune-“hot” and “cold” patients with PDAC/NSCLC are given. (C and D) Correlation analysis between migratory DC population and overall survival across our collected PDAC and NSCLC patient cohorts ( n = 66 patients with PDAC, n = 65 patients with NSCLC). (E and F) ROC curves represent the accuracy of migratory DCs and CD3 + CD8 + T cells in predicting overall survival in patients with PDAC and NSCLC. (G and H) Spatial analysis of multiplex IF images from each group. Magnified images are given. (I and J) Stack bar charts show the density of CD3 + CD8 + T cells at varying distances from migratory DCs within both hot and cold tumors from patients with PDAC/NSCLC. (K) Representative images of multiplex immunostaining of migratory DC and T cell markers on tumor sections from NSCLC immunotherapy non-responder and responder. (L) High population of migratory DCs correlated with better progression-free survival in patients with advanced NSCLC treated with anti-PD-1 ( n = 19 patients). (M) ROC curves illustrate the predictive accuracy of migratory DCs and CD3 + CD8 + T cells for progression-free survival in patients with advanced NSCLC treated with anti-PD-1. (N) Spatial proximity analysis of multiplex IF images from each group. Magnified image is given. (O) Stack bar chart illustrates the density of CD3 + CD8 + T cells at different distances from migratory DCs in each group. (C, D, and L) Log rank (Mantel-Cox). Scale bars in (A), (B), and (K) represent 50 μm. See also Figure S1 and Tables S1 , , and .

Article Snippet: rabbit mAb against human CD3 , CST , Cat# 85061; RRID:AB_2721019.

Techniques: Multiplex Assay, Immunostaining, Derivative Assay

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet: cNP cancer cell @MV DC treatment triggers DC maturation and CD8 + T and NK cell activation in vitro (A and B) FACS analysis measured CD86 expression in immature human/mouse DCs after treatments. Bar charts display the percentage of CD11c + CD86 + DCs per group ( n = 3 independent experiments). (C and D) FACS analysis examined human/murine CD3 + CD8 + T cell proliferation after treatments ( n = 3 independent experiments). (E–H) FACS analysis assessed human/murine CD3 + CD8 + T cell activation after treatments. Bar charts show the percentage of CD3 + CD8 + CD69 + T cells or CD3 + CD8 + IFN-γ + T cells per group ( n = 3 independent experiments). (I–L) FACS analysis assessed human/murine NK cell activation after treatments. Graphs show the percentage of CD3 − CD56 + CD69 + , CD3 − NK1.1 + CD69 + , CD3 − CD56 + IFN-γ + , or CD3 − NK1.1 + IFN-γ + relative to the total human/murine NK population in each group ( n = 3 independent experiments). (M and N) Schematic diagrams illustrate LDH release assays conducted on PDAC patient-derived tumor organoids (T, target), using varying ratios of CD3 + CD8 + T cells (E, effector) pre-treated with the indicated treatments (M), or co-cultured with hDCs pre-treated with the indicated treatments (N). Bar charts indicate the killing efficiency percentage at each E:T ratio among different groups ( n = 3 independent experiments). (O and P) Cytotoxicity LDH release assays on DT6066 cells were conducted using different ratios of CD3 + CD8 + T cells that had been either pre-treated with the indicated treatments (O) or co-cultured with DCs pre-treated with the indicated treatments (P) ( n = 3 independent experiments). (A–L) One-way ANOVA. (M–P) Two-way ANOVA. See also Figure S3 .

Article Snippet: rabbit mAb against human CD3 , CST , Cat# 85061; RRID:AB_2721019.

Techniques: Activation Assay, In Vitro, Expressing, Derivative Assay, Cell Culture

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet: cNP DT6066 @MV DC treatment converts immune-cold pancreatic tumors into hot tumors, reduces hypoxia, and enhances blood vessel function (A) FACS analysis of CD11c + CD86 + DCs in orthotopic DT6066 pancreatic tumors after treatments, with a bar chart showing their percentage relative to total lymphocytes ( n = 3 mice per group). (B) FACS analysis of CD3 + CD4 + CD69 + T cells in orthotopic pancreatic tumors after treatments, with a bar chart showing their percentage relative to total CD3 + CD4 + T cells in each group ( n = 3 mice per group). (C) FACS analysis of CD3 + CD8 + CD69 + T cells in orthotopic pancreatic tumors after treatments, with a bar chart showing their percentage relative to total CD3 + CD8 + T cells in each group ( n = 3 mice per group). (D) Representative images of CD11c and CD86 co-immunostaining in tumor sections from each treatment group. Bar chart shows the quantification of CD11c + CD86 + DC × 10 3 per cm 2 in each group ( n = 3 mice per group). (E and F) Representative images of co-immunostaining for CD3 and CD4 (E) or CD8 (F) in tumor sections from each treatment group ( n = 3 mice per group). (G) FACS analysis of CD3 − NK1.1 + NK cells or CD3 − NK1.1 + CD69 + NK cells in orthotopic pancreatic tumors after treatments. Bar charts show their percentage relative to total CD3 − cells (left) or CD3 − NK1.1 + NK cells (right) in each group ( n = 3 mice per group). (H–J) Cytotoxicity LDH release assays of DT6066 cells after co-culture with various ratios of CD3 + CD8 + T cells isolated from tumors (H), lymph nodes (I), or spleens (J) of orthotopic DT6066 pancreatic tumor-bearing mice after treatments ( n = 3 mice per group). (K) FACS analysis of migratory DCs in orthotopic pancreatic tumors and TDLNs from mice treated as indicated. Bar charts show CD86 + CD103 + DCs as a percentage of CD11c + DCs in each group ( n = 3 mice per group). (L) Representative images of CD11c/CD86/CD103 triple immunostaining on paired tumor and TDLN sections in each group. Bar chart shows the percentage of CD11c + CD86 + CD103 + migratory DC × 10 3 per cm 2 in tumors (left) or TDLNs (right) from each group ( n = 3 mice per group). (M–O) Representative microbubble contrast ultrasound images are shown, and bar charts show quantification across the entire tumors, including tumor cores ( n = 3 mice per group). (P) Representative IHC staining of endomucin in tumor sections from each treatment group. Bar chart shows blood vessel diameter (left) or number of blood vessels (right) per cm 2 in each group ( n = 3 mice per group). (Q) Representative image of GLUT1 and endomucin co-immunostaining in tumor sections from each group. Bar chart shows the relative GLUT1 intensity per group ( n = 3 mice per group). (A–G, K, L, and N–Q) One-way ANOVA. (H–J) Two-way ANOVA. Scale bars in (D), (E), (F), and (L) represent 50 μm. (M) 1 cm. (P and Q) 100 μm. See also Figures S5 and .

Article Snippet: rabbit mAb against human CD3 , CST , Cat# 85061; RRID:AB_2721019.

Techniques: Immunostaining, Co-Culture Assay, Isolation, Triple Immunostaining, Immunohistochemistry

Journal: Cell Reports Medicine

Article Title: Mitochondrial DNA-boosted dendritic cell-based nanovaccination triggers antitumor immunity in lung and pancreatic cancers

doi: 10.1016/j.xcrm.2024.101648

Figure Lengend Snippet:

Article Snippet: rabbit mAb against human CD3 , CST , Cat# 85061; RRID:AB_2721019.

Techniques: Virus, Recombinant, Negative Staining, Lysis, Control, Phospho-proteomics, Cell Isolation, DNA Purification, DNA Extraction, Bicinchoninic Acid Protein Assay, Protein Purification, Magnetic Beads, Software

Journal: Cancer Gene Therapy

Article Title: RevCAR-expressing immune effector cells for targeting of Fn14-positive glioblastoma

doi: 10.1038/s41417-024-00766-8

Figure Lengend Snippet: A Either RevCAR T cells or RevCAR NK-92 cells express reverse chimeric antigen receptors (RevCARs) that consist of an intracellular CD3z signaling domain (SD) as well as a CD28 co-stimulatory domain (CSD), in addition to a CD28 transmembrane and hinge region linked to an extracellular peptide epitope (either E5B9 or E7B6). RevCAR immune cells can be activated by RevTMs. B The novel designed RevTMs are bsAbs derived from the variable light (V L ) and heavy (V H ) chain domains of mAbs directed against either the TAA Fn14 or the RevCAR epitope E5B9 or E7B6. All domains are connected via peptide linkers (Li). Both RevTMs contain an N-terminal signal peptide (SP) and C-terminal Histidines (His) as well as a Strep-tag for their purification. After purification via their Strep tag, the RevTMs were separated by SDS-PAGE followed by ( C ) Coomassie Blue staining, or ( D ) immunoblotting on nitrocellulose membrane and detection via an anti-His mAb and an alkaline phophatase-conjugated anti-mouse Ab. Sor; sortase recognition site, M; molecular weight marker, BSA Std.; bovine serum albumin standard.

Article Snippet: Isolated T cells were stained with fluorescently labeled mAbs against human CD3 (#130-113-138), CD4 (#130-113-225), CD8 (#130-110-683) (Miltenyi Biotec).

Techniques: Derivative Assay, Strep-tag, Purification, SDS Page, Staining, Western Blot, Membrane, Molecular Weight, Marker

Journal: Cancer Gene Therapy

Article Title: RevCAR-expressing immune effector cells for targeting of Fn14-positive glioblastoma

doi: 10.1038/s41417-024-00766-8

Figure Lengend Snippet: A Fn14 expression on luciferase (Luc)-expressing U251 Luc and U343 Luc GBM cells was determined by staining with anti-Fn14 mAb (primary Ab) and AlexaFluor647-conjugated anti-mouse IgG (secondary Ab). The number of Fn14 antigens expressed per cell was detected by a bead-based flow cytometry assay (QIFIKIT). The binding of Fn14-specific RevTMs to Fn14 on both U251 Luc and U343 Luc cell lines was tested using APC-conjugated anti-His mAb. The expression of RevCAR receptors (E5B9 or E7B6) on transduced ( B ) T cells or ( C ) NK-92 cells was confirmed by the anti-La mAb La5B9 or La7B6, respectively, and the secondary Ab AlexaFluor647-conjugated anti-mouse IgG. In addition, the number of RevCAR receptors per NK-92 cell was determined. The binding of the Fn14-specific RevTMs to the respective RevCAR T or NK-92 cells was detected using an APC-conjugated anti-His mAb. A , C Quantitative data from three different experiments are shown as mean ± SD. A – C Flow cytometry data are displayed in histograms (light lines: negative control, dark lines: stained cells).

Article Snippet: Isolated T cells were stained with fluorescently labeled mAbs against human CD3 (#130-113-138), CD4 (#130-113-225), CD8 (#130-110-683) (Miltenyi Biotec).

Techniques: Expressing, Luciferase, Staining, Flow Cytometry, Binding Assay, Negative Control