anti egfr  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: Tailored strategies for improved control of CAR-T cells in multiple myeloma

doi: 10.3389/fimmu.2026.1740345

Figure Lengend Snippet: Efficiency of ADCC and ADC on SLAMF7 and CD19 CAR-T cells in vitro ; (A) Schematic depiction of EGFR-specific ADCC; (B, C) CD19 and SLAMF7 CAR-T cells (CD4:CD8 1:1) were incubated with EGFR antibody cetuximab [50 µg/ml] and cocultured with effector cells (CFSE stained, PBMC w./w.o. NK cells, E:T 50:1 and NK cells, E:T 10:1) for 24 h, CAR-T cell elimination through ADCC is dependent on NK cell availability and CAR-T cell construct, Representative Flow cytometry plots (B) and statistical analysis ((C) n=3) are shown; (D) Lymphocyte (n=12) and NK cell (n=8) counts are reduced after lymphodepleting chemotherapy (d-5, -4 and -3) and only gradually recover; (E) Schematic depiction of the ADC assay; (F) BCMA expression of targeted cells via flow cytometry, (G, H) Cells expressing BCMA or not, were incubated with BCMA targeting ADC belantamab-mafodotin [50 µg/ml]; SLAMF7 CAR-T cells, UTD ctrl T cells (72 h; (G) ) and the human MM cell line OPM-2 (48 h & 72 h; (H) ) are shown; Control antibody: Human IgG1 Isotype Control; n=3 independent donors; Each experiment was performed in triplicates; Two-way ANOVA statistic test was performed using GraphPad Prism 9; Abbreviations: ADC, Antibody-drug conjugate; ADCC, Antibody-dependent cell cytotoxicity; ALC, Absolute Lymphocyte Count; CAR, Chimeric antigen receptor; Ctrl, Control; MM, Multiple myeloma; SLAMF7, SLAM Family member 7 (CD319); UTD, Untransduced. **** = P ≤ 0,0001; *** = P ≤ 0,001; * = P ≤ 0,05.

Article Snippet: Antibodies used in this research report were specific for BCMA-Antibodies used in this research report were specific for BCMA (Miltenyi Biotec; 130-119-152), EGFR (Cetuximab, in-house labelled), CD8 (Miltenyi Biotec; 130-110-683) and CD4 (Miltenyi Biotec; 130-114-534).

Techniques: In Vitro, Incubation, Staining, Construct, Flow Cytometry, Expressing, Control

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Immunological effects of amivantamab in EGFR or MET-expressing non-small cell lung cancer

doi: 10.1007/s00262-026-04369-0

Figure Lengend Snippet: Ex vivo activation of tumor-infiltrating lymphocytes (TILs) by amivantamab according to epidermal growth factor receptor ( EGFR ) mutation status Ex vivo TIL assays were performed as described in Fig. A. Summaries of programmed cell death-1 (PD-1) expression ( A ), interferon-γ (IFNγ) production ( B ), and tumor necrosis factor-α (TNFα) production ( C ) in tumor-infiltrating CD8⁺ T cells, CD80 ( D ) and CD86 expression ( E ) in tumor-infiltrating dendritic cells (DCs), and PD-1 ( F ) and inducible T-cell co-stimulator (ICOS) expression ( G ) in tumor-infiltrating effector regulatory T cells (eTregs) according to EGFR mutation status (left, wild-type (WT); right, mutant) are shown. Statistical analyses were performed Wilcoxon signed-rank sum tests in ( A – G ). In box-and-whisker plots, the box spans from the first to the third quartile with a line at the median and the whiskers extend from the minimum to the maximum. Each dot represents an individual sample and connecting lines indicate paired before-after measurements for the same sample. NS: not significant; * P < 0.05;. ** P < 0.01

Article Snippet: After blocking endogenous peroxidase activity and nonspecific binding, sections were incubated overnight at 4 °C with an anti–human CD8 monoclonal antibody (mAb) (Cell Signaling Technology, Danvers, MA, RRID: AB_2800052), an anti–human CD11c mAb (Cell Signaling Technology, RRID: AB_2799286), an anti–human EGFR mAb (Cell Signaling Technology, RRID: AB_2246311), an anti–human cMET mAb (Cell Signaling Technology, RRID: AB_10858224), an anti-human phospho-EGFR (Tyr1068) mAb (Cell Signaling Technology, RRID:AB_2096270), or an anti-human phospho- Met (Tyr1234/1235) mAb (Cell Signaling Technology, RRID:AB_2143884) diluted in 5% BSA (Albumin, Bovine Serum, F-V, pH5.2; Nacalai tesque, Cat# 01863–48).

Techniques: Ex Vivo, Activation Assay, Mutagenesis, Expressing, Whisker Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Immunological effects of amivantamab in EGFR or MET-expressing non-small cell lung cancer

doi: 10.1007/s00262-026-04369-0

Figure Lengend Snippet: Ex vivo activation of tumor-infiltrating lymphocytes (TILs) by amivantamab according to epidermal growth factor receptor (EGFR) or MET proto-oncogene, receptor tyrosine kinase (MET) expression level ( A ) Distribution of EGFR protein expression levels in tumor samples. EGFR expression was scored by immunohistochemistry (IHC) as follows: 1 + for weak membrane staining in ≥ 10% of tumor cells, 2 + for moderate staining in ≥ 10%, and 3 + for strong staining in ≥ 10%. A pie chart is shown. ( B ) Association between EGFR mutation status and EGFR protein expression. A bar graph is shown. ( C – E ) Tumor-infiltrating CD8⁺ T-cell activation stratified by EGFR expression level. Ex vivo TIL assays were performed as described in Fig. A. Summaries of programmed cell death-1 (PD-1) expression ( C ), interferon-γ (IFNγ) production ( D ), and tumor necrosis factor-α (TNFα) production ( E ) in tumor-infiltrating CD8⁺ T cells according to EGFR expression level (left, ≤ 1 + ; right, ≥ 2 +) are shown. ( F ) Distribution of MET protein expression levels in tumor samples. MET expression was scored by IHC as follows: 1 + for weak membrane staining in ≥ 50% of tumor cells, 2 + for moderate staining in ≥ 50%, and 3 + for strong staining in ≥ 50%. A pie chart is shown. ( G – I ) Tumor-infiltrating CD8⁺ T-cell activation stratified by MET expression level. Ex vivo TIL assays were performed as described in Fig. A. Summaries of PD-1 expression ( G ), IFNγ production ( H ), and TNFα production ( I ) in tumor-infiltrating CD8⁺ T cells according to MET expression level (left, ≤ 1 + ; right, ≥ 2 +) are shown. The correlation between EGFR expression levels (≤ 1 + and ≥ 2 +) and EGFR mutation status was analyzed using Fisher’s exact test in ( B ). Wilcoxon signed-rank sum tests were performed in ( C – E ) and ( G – I ). In box-and-whisker plots, the box spans from the first to the third quartile with a line at the median and the whiskers extend from the minimum to the maximum. Each dot represents an individual sample and connecting lines indicate paired before-after measurements for the same sample. NS: not significant; * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: After blocking endogenous peroxidase activity and nonspecific binding, sections were incubated overnight at 4 °C with an anti–human CD8 monoclonal antibody (mAb) (Cell Signaling Technology, Danvers, MA, RRID: AB_2800052), an anti–human CD11c mAb (Cell Signaling Technology, RRID: AB_2799286), an anti–human EGFR mAb (Cell Signaling Technology, RRID: AB_2246311), an anti–human cMET mAb (Cell Signaling Technology, RRID: AB_10858224), an anti-human phospho-EGFR (Tyr1068) mAb (Cell Signaling Technology, RRID:AB_2096270), or an anti-human phospho- Met (Tyr1234/1235) mAb (Cell Signaling Technology, RRID:AB_2143884) diluted in 5% BSA (Albumin, Bovine Serum, F-V, pH5.2; Nacalai tesque, Cat# 01863–48).

Techniques: Ex Vivo, Activation Assay, Expressing, Immunohistochemistry, Membrane, Staining, Mutagenesis, Whisker Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Immunological effects of amivantamab in EGFR or MET-expressing non-small cell lung cancer

doi: 10.1007/s00262-026-04369-0

Figure Lengend Snippet: Ex vivo activation of tumor-infiltrating lymphocytes (TILs) by amivantamab according to epidermal growth factor receptor (EGFR)/MET proto-oncogene, receptor tyrosine kinase (MET) expression level ( A ) Distribution of EGFR/MET protein expression levels in tumor samples. The EGFR/MET expression was stratified as described in Fig. . A pie chart is shown. ( B – D ) Tumor-infiltrating CD8⁺ T-cell activation stratified by EGFR/MET expression level categorized as group I (both ≤ 1 +) or group II (either ≥ 2 +). Ex vivo TIL assays were performed as described in Fig. A. Summaries of programmed cell death-1 (PD-1) expression ( B ), interferon-γ (IFNγ) production ( C ), and tumor necrosis factor-α (TNFα) production ( D ) in tumor-infiltrating CD8⁺ T cells according to EGFR/MET expression level (left, group I; right, group II) are shown. Statistical analyses were performed by Wilcoxon signed-rank sum tests in ( B – D ). In box-and-whisker plots, the box spans from the first to the third quartile with a line at the median and the whiskers extend from the minimum to the maximum. Each dot represents an individual sample and connecting lines indicate paired before-after measurements for the same sample. NS: not significant; * P < 0.05; *** P < 0.001

Article Snippet: After blocking endogenous peroxidase activity and nonspecific binding, sections were incubated overnight at 4 °C with an anti–human CD8 monoclonal antibody (mAb) (Cell Signaling Technology, Danvers, MA, RRID: AB_2800052), an anti–human CD11c mAb (Cell Signaling Technology, RRID: AB_2799286), an anti–human EGFR mAb (Cell Signaling Technology, RRID: AB_2246311), an anti–human cMET mAb (Cell Signaling Technology, RRID: AB_10858224), an anti-human phospho-EGFR (Tyr1068) mAb (Cell Signaling Technology, RRID:AB_2096270), or an anti-human phospho- Met (Tyr1234/1235) mAb (Cell Signaling Technology, RRID:AB_2143884) diluted in 5% BSA (Albumin, Bovine Serum, F-V, pH5.2; Nacalai tesque, Cat# 01863–48).

Techniques: Ex Vivo, Activation Assay, Expressing, Whisker Assay

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Immunological effects of amivantamab in EGFR or MET-expressing non-small cell lung cancer

doi: 10.1007/s00262-026-04369-0

Figure Lengend Snippet: Ex vivo activation of tumor-infiltrating lymphocytes (TILs) by amivantamab based on epidermal growth factor receptor (EGFR)/MET proto-oncogene, receptor tyrosine kinase (MET) expression level in EGFR - wild type (WT) tumors ( A ) Association between EGFR mutation status and EGFR/MET protein expression. The EGFR/MET expression was stratified as described in Fig. . A bar graph is shown. ( B – D ) Tumor-infiltrating CD8⁺ T-cell activation stratified by EGFR/MET expression level categorized as described in Fig. in EGFR -WT tumors. Ex vivo TIL assays were performed as described in Fig. A. Summaries of programmed cell death-1 (PD-1) expression ( B ), interferon-γ (IFNγ) production ( C ), and tumor necrosis factor-α (TNFα) production ( D ) in tumor-infiltrating CD8⁺ T cells according to EGFR/MET expression level (left, group I; right, group II) are shown. The correlation between EGFR mutation status and EGFR/MET expression classified into group I and group II was analyzed using Fisher’s exact test in ( A ). Wilcoxon signed-rank sum tests were performed in ( B – D ). In box-and-whisker plots, the box spans from the first to the third quartile with a line at the median and the whiskers extend from the minimum to the maximum. Each dot represents an individual sample and connecting lines indicate paired before-after measurements for the same sample. NS: not significant; * P < 0.05

Article Snippet: After blocking endogenous peroxidase activity and nonspecific binding, sections were incubated overnight at 4 °C with an anti–human CD8 monoclonal antibody (mAb) (Cell Signaling Technology, Danvers, MA, RRID: AB_2800052), an anti–human CD11c mAb (Cell Signaling Technology, RRID: AB_2799286), an anti–human EGFR mAb (Cell Signaling Technology, RRID: AB_2246311), an anti–human cMET mAb (Cell Signaling Technology, RRID: AB_10858224), an anti-human phospho-EGFR (Tyr1068) mAb (Cell Signaling Technology, RRID:AB_2096270), or an anti-human phospho- Met (Tyr1234/1235) mAb (Cell Signaling Technology, RRID:AB_2143884) diluted in 5% BSA (Albumin, Bovine Serum, F-V, pH5.2; Nacalai tesque, Cat# 01863–48).

Techniques: Ex Vivo, Activation Assay, Expressing, Mutagenesis, Whisker Assay

Journal: bioRxiv

Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

doi: 10.64898/2026.03.06.710083

Figure Lengend Snippet: A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. B) L-SIGN or DC-SIGN were expressed in Jurkat or primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of primary T cells to authentic Ebola and Sudan virus infection.

Article Snippet: Cells were infected with pseudotyped lentiviruses, and 72 hours later, were harvested and stained with antibodies against the relevant surface marker: mouse CD19 (Miltenyi Biotec, 130-111-884), mouse H2Kk (Miltenyi Biotec, 130-117-235), or human EGFR (R&D Systems, FAB9577R-100), and viability staining was performed using DAPI (Thermo Fisher Scientific, D1306).

Techniques: Expressing, Control, Flow Cytometry, Infection, Mutagenesis, Virus

Journal: bioRxiv

Article Title: Elucidating genes sufficient for viral entry into cells through sequential genome-wide CRISPR activation screens

doi: 10.64898/2026.03.06.710083

Figure Lengend Snippet: A) NGFR was expressed in Jurkat C6 cells using CRISPRa, or alternatively, Jurkat cells using cDNA expression. NGFR -expressing or control cells were then inoculated with rabies pseudovirus encoding mCD19t. Flow cytometry was then performed to determine the percentage of infected cells. This revealed that NGFR expression significantly increased the susceptibility of Jurkat cells to rabies pseudovirus infection. As positive controls, flow cytometry was used to confirm successful delivery of the sgRNA construct as part of the CRISPRa workflow (as denoted by BFP expression) and that NGFR was expressed (upon cDNA expression). B) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression. L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with rabies pseudovirus encoding hEGFRt (a truncated mutant of human EGFR). Flow cytometry was then performed to determine the percentage of infected cells. This revealed that L-SIGN or DC-SIGN expression significantly increased the susceptibility of Jurkat cells and primary T cells to Ebola pseudovirus infection. Cells expressing the highest levels of L-SIGN and DC-SIGN were preferentially infected by Ebola pseudovirus. C) L-SIGN or DC-SIGN were expressed in primary human CD4 + T cells using cDNA expression, and then L-SIGN -expressing, DC-SIGN -expressing, or control cells were inoculated with GFP -expressing Ebola virus or zsGreen -expressing Sudan virus under BSL4 containment. On days 0, 1, and 2 post-infection, flow cytometry was performed to determine the percentage of infected cells and qPCR was performed on cell culture supernatants to quantify viral genome replication. This revealed that L-SIGN or DC-SIGN expression enabled authentic Ebola and Sudan virus entry into primary human T cells, but viral genome replication was impaired, perhaps reflective of cell-intrinsic restriction factors.

Article Snippet: Cells were infected with pseudotyped lentiviruses, and 72 hours later, were harvested and stained with antibodies against the relevant surface marker: mouse CD19 (Miltenyi Biotec, 130-111-884), mouse H2Kk (Miltenyi Biotec, 130-117-235), or human EGFR (R&D Systems, FAB9577R-100), and viability staining was performed using DAPI (Thermo Fisher Scientific, D1306).

Techniques: Expressing, Control, Flow Cytometry, Infection, Construct, Mutagenesis, Virus, Cell Culture