Journal: Cell Death & Disease

Article Title: PD-1 protects expanding human T cells from premature restimulation-induced cell death by modulating TCR and CD28 signaling

doi: 10.1038/s41419-026-08530-6

Figure Lengend Snippet: T cells isolated from healthy human donors were activated with anti-CD3/28 beads and IL-2 was added on Day 3. A PD-1, PD-L1, and PD-L2 expression were measured via flow cytometry over 15 days for N = 8 in CD4+ and N = 5 in CD8 + T cells. B Day 4 purified CD8 + T cells were restimulated with autologous irradiated PBMC and anti-CD3 (1 μg/mL) ± anti-CD28 (1 μg/mL) in the presence of control antibody, anti-PD-1, or anti-PD-L1 at 10 μg/mL for 20 hr. Death was assessed via flow cytometry by propidium iodide staining. Single dot correlates with one human donor. Connecting line compares change in untreated and treated samples within the same donor. N = 3. C Diagram illustrating the proxy APC (pAPC) design created with a 1:1:1 ratio of anti-CD3, anti-CD28, and IgG1κ or recombinant human Fc-chimera PD-L1. D Freshly isolated CD4+ or CD8+ human T cells were activated at a 1:1 cell:bead ratio with IgG or PD-L1 beads and monitored via flow cytometry for CD69 expression after 24 hr ( N = 3 CD4 + , N = 5 CD8 + ) and CD25 expression after 72 hr ( N = 3 CD4 + , N = 6 CD8 + ). E Blastogenesis patterns of unactivated, IgG, or PD-L1 activated cells on Day 3 are demonstrated by dot plots assessing forward scatter (FSC) (x-axis) and side scatter (SSC) (y-axis). The progressive changes in FSC monitored via flow cytometry over 3 days are summarized for CD4+ and CD8 + T cells. N = 3 CD4 + , N = 5 CD8 + .

Article Snippet: Isotype control antibodies were paired with the corresponding blocking antibody at 10 μg/mL: IgG1κ (BioXCell, #BP0297) or IgG4 (BioXCell, #CP148).

Techniques: Isolation, Expressing, Flow Cytometry, Purification, Irradiation, Control, Staining, Recombinant

Journal: Cell Death & Disease

Article Title: PD-1 protects expanding human T cells from premature restimulation-induced cell death by modulating TCR and CD28 signaling

doi: 10.1038/s41419-026-08530-6

Figure Lengend Snippet: A CD8+ or CD4 + T cells were restimulated on Day 4 or Day 11 with p328-IgG of p328-L1 beads at a 1:1 bead:cell ratio and monitored for cell loss via flow cytometry 24 hr later using propidium iodide. N = 8 CD4+ early, N = 11 CD4+ late, N = 9 CD8+ early, N = 18 CD8+ late. B The difference in % cell loss between p328-IgG and p328-L1 beads in ( A ) is plotted for each donor, cell type, and cell stage evaluated. (same N as ( A ). C RICD assays set-up in ( A ) were examined on the flow cytometer using propidium iodide to calculate the percentage of live cells remaining after 24 hr of restimulation. N = 7 CD4+ early, N = 9 CD4+ late, N = 12 CD8+ early, N = 8 CD8+ late. D PD-L1 dose response was determined using the standard RICD setup amd a new bead set that gradually decreased PD-L1 concentration on the bead from 33% to 0%. Day 4 CD4+ and CD8 + T cells were utilized in this experiment. N = 12 CD4 + , N = 10 CD8 + , statistical analysis was calculated with two-way ANOVA. E Diagram representing RICD setup and creation of secondary bead set that was ligated 100% to IgG1κ (control secondary bead) or 67% IgG1κ and 33% recombinant PD-L1, matching the PD-L1 protein concentration on the p328-L1 primary bead. F Day 4 CD8+ and CD4 + T cells were restimulated with a 2:1 bead:cell ratio comprising a 1:1 ratio of the primary bead and 1:1 ratio of the secondary bead. RICD was monitored as in ( A ) N = 12.

Article Snippet: Isotype control antibodies were paired with the corresponding blocking antibody at 10 μg/mL: IgG1κ (BioXCell, #BP0297) or IgG4 (BioXCell, #CP148).

Techniques: Flow Cytometry, Concentration Assay, Control, Recombinant, Protein Concentration

Journal: Cell Death & Disease

Article Title: PD-1 protects expanding human T cells from premature restimulation-induced cell death by modulating TCR and CD28 signaling

doi: 10.1038/s41419-026-08530-6

Figure Lengend Snippet: A Representative histogram of data displaying demarcation of AnnexinV hi cells -/+ bead restimulation. B Day 4 and Day 11 CD4+ and CD8 + T cells were left unstimulated or were restimulated with p328-IgG or p328-L1 beads for 6 hr before being stained with AnnexinV and propidium iodide and read on the flow cytometer. N = 6. C Representative flow cytometric histogram of PI cell cycle phases based on DNA content. D Day 4 and Day 11 CD4+ and CD8 + T cells were left unstimulated or were restimulated with p328-IgG or p328-L1 beads for 18 hr before being permeabilized and stained with propidium iodide to determine % of cells in the subG1 phase. N = 8 CD4+ early, N = 8 CD4+ late, N = 10 CD8+ early, N = 9 CD8+ late. E Analysis using gated cells from ( D ) was performed to quantify changes in the G1 phase. F Cell phase characterization from ( D ) was analyzed to determine the PD-L1 induced difference in subG1 and G1 phases.

Article Snippet: Isotype control antibodies were paired with the corresponding blocking antibody at 10 μg/mL: IgG1κ (BioXCell, #BP0297) or IgG4 (BioXCell, #CP148).

Techniques: Staining, Flow Cytometry

Journal: Cell Death & Disease

Article Title: PD-1 protects expanding human T cells from premature restimulation-induced cell death by modulating TCR and CD28 signaling

doi: 10.1038/s41419-026-08530-6

Figure Lengend Snippet: A CD8+ or CD4 + T cells were restimulated on Day 4 with soluble anti-CD3 (OKT3) at 100 ng/mL or anti-CD3/CD28 coated beads at a 1:1 bead:cell ratio. Cell loss/apoptosis was quantified 18–24 hr later after propidium iodide staining and flow cytometry. N = 5 CD4+ early, N = 6 CD4+ late, N = 5 CD8+ early, N = 6 CD8+ late. B Day 4 and Day 11 CD4+ and CD8 + T cells from N = 5 individual donors were stained for CD28 and PD-1 expression and compared via flow cytometry. C Diagram illustrating restimulation beads conjugated with or without anti-CD28, with p328-IgG used to maintain total protein concentration. D CD8+ or CD4 + T cells were restimulated on Day 4 with beads illustrated in ( C ) at a 1:1 bead:cell ratio, monitored for cell loss 24 hr later using propidium iodide using flow cytometry. E The difference in cell loss induced by restimulation with beads -/+ anti-CD28 (leaving anti-CD3 + /- PD-L1-Fc constant) was plotted for every donor ( N = 10).

Article Snippet: Isotype control antibodies were paired with the corresponding blocking antibody at 10 μg/mL: IgG1κ (BioXCell, #BP0297) or IgG4 (BioXCell, #CP148).

Techniques: Staining, Flow Cytometry, Expressing, Protein Concentration

Journal: Cell Death & Disease

Article Title: PD-1 protects expanding human T cells from premature restimulation-induced cell death by modulating TCR and CD28 signaling

doi: 10.1038/s41419-026-08530-6

Figure Lengend Snippet: A Day 4 CD4+ and CD8 + T cells ( N = 4 donors each) were restimulated with p328-IgG or p328-L1 beads at a 1:1 bead:cell ratio for 5 hr prior to lysate collection. Control cells were left unstimulated or treated with 1 μM STX. After cell lysis and protein standardization, donor lysates were pooled for each stimulation condition and analyzed on the apoptosis protein array. Representative spots for differentially expressed proteins are shown. Protein signal intensity from membrane imaging was analyzed and normalized using QuickSpots software. T cells were restimulated as in ( A ), lysed in RIPA buffer and separated by SDS-PAGE to quantify FASL ( B ) and survivin ( C ). Data are representative of experiments for 3–4 separate donors. ( D ) Day 4 CD4+ and CD8 + T cells were treated with 10 μg/mL anti-FASL blocking antibody for 1 hr prior to restimulation with the quad bead set. Cells were restimulated for 24 hr and stained with propidium iodide; % viability was assessed by flow cytometry. N = 7 CD4 + , N = 5 CD8 + .

Article Snippet: Isotype control antibodies were paired with the corresponding blocking antibody at 10 μg/mL: IgG1κ (BioXCell, #BP0297) or IgG4 (BioXCell, #CP148).

Techniques: Control, Lysis, Protein Array, Membrane, Imaging, Software, SDS Page, Blocking Assay, Staining, Flow Cytometry

Journal: medRxiv

Article Title: Inflammatory arthritis immune related adverse events represent a unique autoimmune disease entity primarily driven by T cells, but likely not autoantibodies

doi: 10.1101/2025.06.06.25328991

Figure Lengend Snippet: Humoral immunity remains largely intact in irAE patients, while seronegative RA patients have increased atypical B cell frequency and autoantibodies. (A) Expression of IgM and IgD on CD27 - CD24 - CD38 - B cells. Right, percentage of IgD + CD27 - CD24 - CD38 - naïve B cells and IgM - IgD - CD27 - CD24 - CD38 - double-negative B cells. (B) Expression of CD27 and CD38 on CD19 + B cells. Right, percentages of CD27 hi CD38 hi antibody-secreting cells (ASCs) and CD27 + CD38 - memory B cells. (C) Expression of CD138 on CD27 hi CD38 hi ASCs. Right, percentage of CD138 + ASCs and CD138 - ASCs. (D) Expression of CD11c and CD21 on CD19 + B cells. Right, percentage of CD11c + CD21 - CD19 + B cells. A-D, HC (n = 63), irAE (n = 33), RAC (n = 46) ICI, (n = 20). (E-G) GSEA was performed on the B cells between irAE and ICI. (E) Significantly enriched pathways in B cells from irAE and ICI. GSEA plots of IFN-α and IFN-γ response (F), and oxidative phosphorylation (G). (H) The volcano plot of the citrullinated or non-citrullinated relative IgG or IgM-isotype autoantigen levels comparing RA vs HC, irAE vs ICI, irAE vs HC, or irAE vs RA. The autoantigens were labeled when p < 0.01. (I) Immunoglobulin isotype levels in the plasma were measured by multiplex assay, HC (n = 22), irAE (n = 34), RAC (n = 46), ICI (n = 26). (J) CXCL13 levels in the plasma were measured by ELISA, HC (n = 20), irAE (n = 34) RAC (n = 47), ICI (n = 23). (K) BAFF levels in the plasma were measured by multiplex assay, HC (n = 21), irAE (n = 34), RAC (n = 47), ICI (n = 18). Data in graphs represent mean ± SEM, Significance was tested by one-way ANOVA (A-D, I-K), logistic regression (H).

Article Snippet: 100 μg/mL pembrolizumab (Keytruda) or Isotype control IgG4 (BioXcell, Cat# CP147) was added into the culture media 2 days after stimulation, and B cells were further cultured for another 5 days.

Techniques: Expressing, Phospho-proteomics, Immunopeptidomics, Labeling, Clinical Proteomics, Multiplex Assay, Enzyme-linked Immunosorbent Assay

Journal: medRxiv

Article Title: Inflammatory arthritis immune related adverse events represent a unique autoimmune disease entity primarily driven by T cells, but likely not autoantibodies

doi: 10.1101/2025.06.06.25328991

Figure Lengend Snippet: Pembrolizumab does not substantially affect B cell activation and antibody production ex vivo (A-C) Human naïve B cells were isolated from PBMCs and cultured under indicated conditions for 7 days; Pembrolizumab or isotype control IgG4 was added on Day 2, n = 8. (A) Expression of CD38 and CD27 on B cells. Right, percentages of CD27 + CD38 - , CD27 + CD38 + B cells. (B) Expression of CD138 on CD27 + CD38 + B cells. Right, percentage of CD138 + CD27 + CD38 + B cells. (C) Different immunoglobulin isotypes were measured in the culture supernatant of (A-B) by multiplex assay. (D-F) B cells from HuPD-1 mice were isolated and cultured with LPS, IL4, BAFF, or ODN2006, anti-IgM, IL-21, IL-4 or R848, anti-IgM, anti-CD40, IL-21, IFN-γ for 3 days, pembrolizumab or isotype control was added on Day 1. (D) Expression of IgG2c on activated B cells. Right, percentage of IgG2c + B cells, n = 5. (E) Expression of IgG1 on activated B cells. Right, percentage of IgG1 + B cells, n = 5. (F) Different immunoglobulin isotypes in the supernatant of were measured by multiplex assay, n = 5. Data in graphs represent mean ± SEM, Significance was tested by Two-way ANOVA.

Article Snippet: 100 μg/mL pembrolizumab (Keytruda) or Isotype control IgG4 (BioXcell, Cat# CP147) was added into the culture media 2 days after stimulation, and B cells were further cultured for another 5 days.

Techniques: Activation Assay, Ex Vivo, Isolation, Cell Culture, Control, Expressing, Multiplex Assay

Journal: medRxiv

Article Title: Inflammatory arthritis immune related adverse events represent a unique autoimmune disease entity primarily driven by T cells, but likely not autoantibodies

doi: 10.1101/2025.06.06.25328991

Figure Lengend Snippet: PD-1 inhibition does not have substantial impact on B cells ex vivo (A) Human naïve B cells were isolated from healthy donor PBMCs and stimulated with different conditions; Pembrolizumab (Keytruda) or isotype control IgG4 was added into the culture media on day 2, mean fluorescence intensity (MFI) of CD86 on B cells was measured, n = 8. (B-F) B cells were isolated from HuPD-1 mice, labeled with CTV, and cultured under 3 different conditions: LPS, rmIL-4, BAFF or Anti-IgM, CpG ODN, rmIL-21, rmIL-4, 100 rhIL-2 or R848, anti-CD40, anti-IgM, rmIL-21, rmIFN-γ for 3 days. Pembrolizumab or isotype control IgG4 was added on day 1, n = 5. (B) Summary of human PD-1 MFI on B cells from different groups. (C) Expression of CD138 on activated B cells. Right, percentage of CD138 + B cells. (D) Summaries of activation marker CD69 and CD86 MFIs on B cells from different groups. (E) Summaries of CD71 and CD98 MFIs on B cells from different groups. (F) Representative flow cytometry plot of CTV dilution on B cells. Right, a summary of the proliferation index of B cells from different groups. Data in graphs represent mean ± SEM, Significance was tested by Two-way ANOVA.

Article Snippet: 100 μg/mL pembrolizumab (Keytruda) or Isotype control IgG4 (BioXcell, Cat# CP147) was added into the culture media 2 days after stimulation, and B cells were further cultured for another 5 days.

Techniques: Inhibition, Ex Vivo, Isolation, Control, Fluorescence, Labeling, Cell Culture, Expressing, Activation Assay, Marker, Flow Cytometry

Journal: medRxiv

Article Title: Inflammatory arthritis immune related adverse events represent a unique autoimmune disease entity primarily driven by T cells, but likely not autoantibodies

doi: 10.1101/2025.06.06.25328991

Figure Lengend Snippet: Inflammatory cytokines enriched in irAE patients promote both CD4 + and CD8 + T cell functions (A) Summaries of the percentage of CD45RA - CD8 + T cells at day 1 or day 5 of the culture, n = 6. (B) Summary of the percentage of Ki67 + in CD45RA - CD8 + T cells, n = 4. (C) Expression of CD69 on cultured CD8 + T cells at day 1. Right, CD69 mean fluorescence intensity (MFI) relative to the control in each experiment, n = 5. (D) Summary of the CD25 MFI relative to the control in each experiment, n = 5. (E) Summary of the percentage of Granzyme B + CD8 + T cells, n = 5. (F) Summary of the percentage of CD45RA - CD4 + T cells at day 5 of the culture, n = 4. (G) Summary of the percentage of Ki67 + CD45RA - CD4 + T cells, n = 4. (H-L) CD4 + T cells were isolated from healthy donor PBMCs, stimulated with 10 μg/mL plate-coated anti-human CD3 and anti-human CD28 in the presence of vehicle control (control), 100 ng/mL IFN-α, 100 ng/mL IL-6, 100 ng/mL IL-12, or the combination of them for 5 days, followed by restimulating with PMA/Ionomycin/Monensin for 5 h. (H) Expression of TNF-α in CD45RA - CD4 + T cells. Right, percentage of TNF-α + cells in CD45RA - CD4 + T cells, n =4. (I) Expression of IL-2 and IL-4 in CD45RA - CD4 + T cells. Right, percentage of IL-4 + and IL-2 + cells in CD45RA - CD4 + T cells, n = 3. (J-L) Summaries of the percentages of TNF-α + (J), IL-4 + (K), or IL-2 + (L) cells in CD38 + CXCR5 - or CD38 - CXCR5 + CD4 + T cells at day 5, n = 4. (M-Q) PBMCs from irAE patients were stimulated with 10 μg/mL plate-coated anti-human CD3 and anti-human CD28 in presence of IgG1 isotype control or a combination of 50 μg/mL anti-human IL-6R, 50 μg/mL anti-human IL-12p40 and 50 μg/mL anti-human IFNAR1 for 3 days, n = 9. (M) Expression of granzyme B and perforin in CD8 + T cells. Right, percentage of granzyme-B + perforin + CD8 + T cells. (N) MFI of MTG in CD8 + T cells. (O, P) MFI of GluCy5 (O), TMRM and MTDR (P), and MTG (Q) in CD4 + T cells. Data in graphs represent mean ± SEM, Significance was tested by One-way ANOVA (A-L), and paired Student’s T-test (N-Q).

Article Snippet: 100 μg/mL pembrolizumab (Keytruda) or Isotype control IgG4 (BioXcell, Cat# CP147) was added into the culture media 2 days after stimulation, and B cells were further cultured for another 5 days.

Techniques: Expressing, Cell Culture, Fluorescence, Control, Isolation

Journal: medRxiv

Article Title: Inflammatory arthritis immune related adverse events represent a unique autoimmune disease entity primarily driven by T cells, but likely not autoantibodies

doi: 10.1101/2025.06.06.25328991

Figure Lengend Snippet: Inflammatory cytokines enriched in irAE patients promote both CD4 + and CD8 + T cell functions (A-G) CD8 + T cells were isolated from healthy donor PBMCs and cultured in the plate coated with 10 μg/mL anti-human CD3 and anti-human CD28 for indicated days in the presence of vehicle control (control), 100 ng/mL IFN-α, 100 ng/mL IL-6, 100 ng/mL IL-12, or the combination of IFN-α, IL-6 and IL-12. (A) Summaries of the percentage of CD38 + CD127 - CD8 + T cells at day 1 and day 5, n = 5. (B) CD69 expression was measured on day 5, and the mean fluorescence intensity (MFI) of CD69 was summarized on the right, n = 4. (C) CD25 expression was measured on day 5, and the MFI of CD25 was summarized on the right, n = 4. (D) CD8 + T cells activated for 5 days were restimulated with PMA/Ionomycin/Monensin for 5 h, expression of granzyme B and IFN-γ in CD8 + T cells were examined by flow cytometry. Right, percentage of granzyme B + IFN-γ + CD8 + T cells, n = 5. (E-G) CD8 + T cells were cultured for 5 days. MFIs (all relative to those in control condition) of MTDR (E), MTG (F), and GluCy5 (G) were summarized, n = 5. (H-L) CD4 + T cells were isolated from the PBMCs of HC, cultured in the plate coated with 10 μg/mL anti-human CD3 and anti-human CD28 for indicated days in the presence of 100 ng/mL IFN-α, 100 ng/mL IL-6, 100 ng/mL IL-12, control or the combination of them. (H) CD4 + T cells activated for 5 days were restimulated with PMA/Ionomycin/Monensin for 5 h, and IL-21 level was examined in the CD45RA - CD4 + T cells, n =3. (I) Activated CD4 + T cells were restimulated with 10 μg/mL plate-coated anti-human CD3 and anti-human CD28 with monensin for 6 h, and expression of CXCL13 was measured in CD45RA - CD4 + T cells, n = 4. (J) CD38 + CXCR5 - or CD38 - CXCR5 + CD4 + T cells were examined at day 5, and the percentages were summarized, n = 4. (K) Percentage of CXCL13 + cells in CD38 + CXCR5 - or CD38 - CXCR5 + CD4 + T cells at day 5, n = 4. (L) Percentage of IL-21 + cells in CD38 + CXCR5 - or CD38 - CXCR5 + CD4 + T cells at day 5, n = 3. (M-S) PBMCs from the irAE patients were cultured in the plate coated with 10 μg/mL anti-CD3 and anti-CD28 in the presence of IgG1 isotype control or 50 μg/mL anti-human IL-6R, 50 μg/mL anti-human IL-12p40 and 50 μg/mL anti-human IFNAR1 for 3 days, n = 9. (M) Expression of CD38 and CD127 on CD8 + T cells. Right, percentage of CD38 + CD127 - CD8 + T cells. (N, O) CD8 + T cells activated for 5 days were restimulated with PMA/Ionomycin/Monensin for 5 h, (N) Expression of Granzyme B and IFN-γ. Right, percentage of granzyme B + IFN-γ + CD8 + T cells, (O) Expression of perforin and IFN-γ. Right, percentage of perforin + IFN-γ + CD8 + T cells. (P, Q) MFIs of GluCy5 (P), TMRM, and MTDR (Q) in CD8 + T cells were presented. (R, S) CD4 + T cells activated for 5 days were restimulated with PMA/Ionomycin/Monensin for 5 h, (R) Expression of IL-21 and IL-2. Right, percentage of IL-21 + IL-2 + CD4 + T cells. (S) Expression of CD38 and CXCR5 on CD4 + T cells. Right, percentage of CD38 + CXCR5 - CD4 + T cells. Data in graphs represent mean ± SEM, Significance was tested by One-way ANOVA (A-L), and paired Student’s t-test (M-S).

Article Snippet: 100 μg/mL pembrolizumab (Keytruda) or Isotype control IgG4 (BioXcell, Cat# CP147) was added into the culture media 2 days after stimulation, and B cells were further cultured for another 5 days.

Techniques: Isolation, Cell Culture, Control, Expressing, Fluorescence, Flow Cytometry

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virus V937 in combination with PD-1 blockade therapy to target immunologically quiescent liver and colorectal cancer

doi: 10.1016/j.omton.2024.200807

Figure Lengend Snippet: Anti-PD-1 treatment of co-culture of CRC cell line or organoid with PBMCs potentiate V937-induced oncolysis (A) Induction of ICAM-1 expression in COLO205 in a non-contact co-culture system with PBMCs upon anti-PD-1 antibody treatment in the presence of 5 ng/mL anti-CD3 for 72 h. (B) COLO205 or co-culture of COLO205 and PBMCs, in presence of anti-CD3 mAb (5 ng/mL) was treated with either IgG4 isotype control (10 μg/mL), anti-PD-1 pembrolizumab (10 μg/mL), V937 (MOI = 5), and pembrolizumab (10 μg/mL) + V937 (MOI = 5) ( n = 4) for 72 h at 37°C, 5% CO 2 . Cell viability of COLO205 was determined in CellTiter-Glo 2.0 assay (Promega). Mean percentages of viability, normalized to Ig4 isotype control-treated, ± standard deviations are shown. p values between V937 and anti-PD-1 + V937 groups are shown; significant differences are indicated by the p values when p < 0.05 (NS = not significant). (C) The media from the co-culture of COLO205 cells and PBMCs with the various treatments were harvested at 72 h and assayed for protein concentrations of various cytokines and chemokines using multiplexed electrochemiluminescence-based assays ( n = 3). Mean ± standard deviations for IFN-α, IFN-γ, IP-10, and IL-1β are shown. p values between V937 and anti-PD-1 + V937 groups are shown; significant differences are indicated by the p values when p < 0.05. (D) Cell viability was assessed in CRC-3 organoids in CellTiter-Glo 2.0 assay following treatment of the co-culture of CRC-3 organoids and PBMCs, in presence of anti-CD3 (5 ng/mL), with either IgG4 isotype control (10 μg/mL), anti-PD-1 pembrolizumab (10 μg/mL), V937 (MOI = 5), and pembrolizumab (10 μg/mL) + V937 (MOI = 5) for 72 h at 37°C, 5% CO 2 ( n = 4). Significant differences are indicated by the p values when p < 0.05. (E) Representative brightfield images of CRC-3 organoids from the co-culture with PBMCs at 72 h under the treatment of IgG4 isotype control, V937 (MOI = 5), pembrolizumab (10 μg/mL), V937 (MOI = 5), and pembrolizumab (10 μg/mL) + V937 (MOI = 5), all treatment groups included the presence of anti-CD3 mAb (5 ng/mL).

Article Snippet: Human IgG4 S228P isotype control (Lot 60AGK), anti-PD-1 hIgG4 S228P Pembrolizumab (Lot 30ADQ) were generated at Merck Research Laboratories.

Techniques: Co-Culture Assay, Expressing, Electrochemiluminescence

Journal: Molecular Therapy Oncology

Article Title: Oncolytic virus V937 in combination with PD-1 blockade therapy to target immunologically quiescent liver and colorectal cancer

doi: 10.1016/j.omton.2024.200807

Figure Lengend Snippet: Intratumoral V937 in combination with pembrolizumab reduced CRC-2 organoid burden in humanized mice CRC-2 organoids (2 × 10 6 cells) in 1:1 suspension with Matrigel were subcutaneously injected into the right flanks of 40 female, 20-week-old CD34+ humanized NSG mice (The Jackson Laboratory). (A) IHC analysis of human ICAM-1 on CRC-2 tumors harvested 42 days following implant is shown. Scale bar, 100 μm. (B) Dosing was initiated 2 weeks following implantation of the CRC-2 organoids when the mean tumor volume reached 80 mm 3 . The 40 humanized mice engrafted with CD34+ cells from three immune donors were distributed among four groups: Group 1: PBS or human IgG4 isotype control (10 mg/kg) ( n = 9), Group 2: V937 (10 8 PFU) ( n = 11), Group 3: Pembrolizumab (10 mg/kg) ( n = 9), and Group 4: V937 (10 8 PFU) + pembrolizumab (10 mg/kg) ( n = 11). PBS and V937 were administered intratumorally on days 0, 2, 4, 6, 13, 17, and 20, and isotype control or pembrolizumab were administered intraperitoneally on days 0, 7, and 14. Mean tumor volumes ± standard error of the mean is shown. (C) Individual tumor growth curves for each animal in the four groups are shown.

Article Snippet: Human IgG4 S228P isotype control (Lot 60AGK), anti-PD-1 hIgG4 S228P Pembrolizumab (Lot 30ADQ) were generated at Merck Research Laboratories.

Techniques: Suspension, Injection