goat anti dkk1  (R&D Systems)

Journal: Nature Communications

Article Title: T cells engineered against Dickkopf-1-A2 complex can be used to treat HLA-A2 + solid and hematologic cancers

doi: 10.1038/s41467-026-69621-8

Figure Lengend Snippet: a Expression of DKK1 mRNA across hematologic tumors was ranked, and datasets of GSE13591 , GSE2350 , and GSE13159 from Oncomine database were used. MM (multiple myeloma), Pro-B-ALL (Pro-B cell acute lymphoblastic leukemia), MGUS (monoclonal gammopathy of undetermined significance), PCL (primary plasma cell leukemia), B-ALL Child (B-Cell acute lymphoblastic leukemia in children), FL (follicular lymphoma), B-ALL (B-cell acute lymphoblastic leukemia), MS (myelodysplastic syndrome), CML (chronic myeloid leukemia). b Expression of DKK1 mRNA across TCGA solid tumors compared to paired normal tissues was ranked in GEPIA2 database. PAAD (pancreatic adenocarcinoma), CHOL (cholangiocarcinoma), ESCA (esophageal carcinoma), HNSC (head and neck squamous cell carcinoma), LUSC (lung squamous cell carcinoma), LIHC (liver hepatocellular carcinoma), STAD (stomach adenocarcinoma), THYM (thymoma), UCS (uterine carcinosarcoma), KIRC (kidney renal clear cell carcinoma), LUAD (lung adenocarcinoma), ACC (adrenocortical carcinoma), CESC (cervical squamous cell carcinoma and endocervical adenocarcinoma). c , d Overall and disease-free survival of pancreatic cancer patients with DKK1 -high versus DKK1 -low expression in GEPIA2 database. The top 50% patients with higher expressions of DKK1 were defined as DKK1 -high, and the rest were defined as DKK1 -low patients. Hazards ratio and p value are demonstrated. Cox proportional hazards regression was used. e Solid tumor datasets of Pei Pancreas ( GSE16515 , normal n = 16, tumor n = 36), Landi Lung ( GSE10072 , normal n = 49, tumor n = 58) and Curtis Breast (EGAS00000000083, normal n = 144, tumor n = 250) from Oncomine database showing the expression of DKK1 mRNA in tumor and paired normal tissues. Two-tailed Student’s t -test was used. f Immunohistochemistry staining of DKK1-A2 complex on normal pancreatic, lung, or breast tissues from 5 randomly selected HLA-A2 positive donors. g–i Immunohistochemistry staining of DKK1, HLA-A2, and DKK1-A2 complex of 10 randomly selected HLA-A2 positive or negative PDAC, NSCLC, and TNBC tumor tissues. The independent experiments were repeated three times, and representative images are showed for immunohistochemistry staining. Data shown as mean ± SEM. Source data are provided as a file.

Article Snippet: Sections were incubated with the primary antibody of DKK1 (Santa Cruz Biotechnology, 10 μg/mL), HLA-A2 (Santa Cruz Biotechnology, 10 μg/mL), DKK1-A2 (mC2/hC2, 10 μg/mL), or CEA (NeoBiotechnologies, 10 μg/mL) overnight at 4 °C.

Techniques: Expressing, Clinical Proteomics, Two Tailed Test, Immunohistochemistry, Staining

Journal: Nature Communications

Article Title: T cells engineered against Dickkopf-1-A2 complex can be used to treat HLA-A2 + solid and hematologic cancers

doi: 10.1038/s41467-026-69621-8

Figure Lengend Snippet: a Schematic presentation showing the recognition of DKK1-A2 CAR-T cells and DKK1-A2 expressing tumor cells. b Schematic presentation showing DKK1-A2 and CD19 CAR constructs. c Percentage of CAR expressing T cells determined as eGFP positive T cells by flow cytometry analysis. d Expression of DKK1, HLA-A2 or DKK1-A2 complex by JeKo-1, Daudi, U266, KMS-26, CFPAC-1, PANC-1, NCI-H441 or MDA-MB-231 tumor cell lines determined by flow cytometry. e CAR-T cell proliferation measured by dilution of Celltrace Violet analyzed by flow cytometry. CAR-T cells were stained with Celltrace Violet and co-cultured with JeKo-1, Daudi, U266 or CFPAC-1 tumor cells at an E:T ratio of 1:1 for 72 h. f Cytotoxicity of CAR-T cells determined by luciferase reporter assay. CAR-T cells were co-cultured with Luc-transduced JeKo-1, Daudi, U266, KMS-26, CFPAC-1, PANC-1, NCI-H441, or MDA-MB-231 tumor cells at indicated E:T ratios for 12 h. n = 3, technical replicates. g , h Percentages of IFN-γ and TNF-α producing CD4 + or CD8 + CAR-T cells determined by intracellular cytokine staining. CAR-T cells were co-cultured with JeKo-1, Daudi, MM.1S-A2, KMS-12-BM-A2, CFPAC-1 or PANC-1 tumor cells at an E:T ratio of 1:1 overnight. In the experiments, CD19 CAR-T cells were used as a control. n = 3, technical replicates. Data shown as mean ± SEM. Source data are provided as a file.

Article Snippet: Sections were incubated with the primary antibody of DKK1 (Santa Cruz Biotechnology, 10 μg/mL), HLA-A2 (Santa Cruz Biotechnology, 10 μg/mL), DKK1-A2 (mC2/hC2, 10 μg/mL), or CEA (NeoBiotechnologies, 10 μg/mL) overnight at 4 °C.

Techniques: Expressing, Construct, Flow Cytometry, Staining, Cell Culture, Luciferase, Reporter Assay, Control

Journal: Nature Communications

Article Title: T cells engineered against Dickkopf-1-A2 complex can be used to treat HLA-A2 + solid and hematologic cancers

doi: 10.1038/s41467-026-69621-8

Figure Lengend Snippet: a Expression of DKK1, HLA-A2, and DKK1-A2 complex by primary tumor cells of 7 randomly selected PDAC patients determined by flow cytometry. b CAR-T cell proliferation measured by dilution of Celltrace Violet, analyzed by flow cytometry. CAR-T cells were stained with Celltrace Violet and co-cultured with primary PDAC tumor cells from the patients at an E:T ratio of 1:1. c Representative flow staining, and d Summarized results depicting the in vitro cytolytic activity of CAR-T cells against primary PDAC cells determined by flow cytometry, measuring the number or percentage of primary tumor cells and CAR-T cells in their co-culture. Primary PDAC tumor cells were stained with Celltrace Violet and co-cultured with CAR-T cells for 72 h. n = 2, technical replicates. One-way ANOVA was used. e – g Percentages of IFN-γ, TNF-α, and Granzyme B producing cells determined by intracellular cytokine staining. CAR-T cells were co-cultured with primary PDAC tumor cells at an E:T ratio of 1:1 overnight. n = 4, biological replicates. Two-tailed Student’s t -test was used. h Schema of PDAC PDX model treated with CAR-T cells, i Tumor volume of PDAC PDX mice measured weekly. Two-way ANOVA was used, j Survival of PDAC PDX mice monitored daily, Log-rank test was used, and k Percentages of CAR-T cells in blood, spleen, and bone marrow in PDAC PDX mice identified as GFP expression by flow cytometry before and post-CAR-T cell infusion. Percentage (%) was defined as the proportion of eGFP + CAR-T cells in blood cells, splenocytes, or bone marrow cells. Ctrl, n = 5; CD19 CAR-T, n = 5; mC2 CAR-T, n = 5; biological replicates. Two-tailed Student’s t -test was used. In some experiments, CD19 CAR-T cells were used as a control. Data shown as mean ± SEM. Source data are provided as a file.

Article Snippet: Sections were incubated with the primary antibody of DKK1 (Santa Cruz Biotechnology, 10 μg/mL), HLA-A2 (Santa Cruz Biotechnology, 10 μg/mL), DKK1-A2 (mC2/hC2, 10 μg/mL), or CEA (NeoBiotechnologies, 10 μg/mL) overnight at 4 °C.

Techniques: Expressing, Flow Cytometry, Staining, Cell Culture, In Vitro, Activity Assay, Co-Culture Assay, Two Tailed Test, Control

Journal: Nature Communications

Article Title: T cells engineered against Dickkopf-1-A2 complex can be used to treat HLA-A2 + solid and hematologic cancers

doi: 10.1038/s41467-026-69621-8

Figure Lengend Snippet: a Immunohistochemistry staining of human normal tissue microarray containing 35 different types of normal tissues (including Marker) from three HLA-A2 positive donors stained with DKK1, HLA-A2 and DKK1-A2 complex. b , c Levels of secreted human IFN-γ and IL-6 in co-culture of CAR-T cells with autologous monocytes and JeKo-1 tumor cells for 24 h determined by ELISA. n = 5, technical replicates. One-way ANOVA was used. d , e Levels of secreted human IFN-γ and murine IL-6 in the peritoneal lavage fluid of JeKo-1 bearing mice treated with intraperitoneally infused an excessive dose of CAR-T cells determined 3 days post CAR-T cell infusion. n = 5, biological replicates. One-way ANOVA was used. f Body weight change of SCID mice transferred with the excessive dose of CAR-T cells monitored every 24 h. n = 5, biological replicates. Two-way ANOVA was used. g – i Numbers of immune cell populations in blood, spleen and bone marrow of hDKK1-A2 mice treated with murine DKK1-CAR-T cells analyzed by flow cytometry. n = 5, biological replicates. One-way ANOVA was used. In most experiments, CD19 CAR-T cells were used as a control. Data shown as mean ± SEM. Source data are provided as a file.

Article Snippet: Sections were incubated with the primary antibody of DKK1 (Santa Cruz Biotechnology, 10 μg/mL), HLA-A2 (Santa Cruz Biotechnology, 10 μg/mL), DKK1-A2 (mC2/hC2, 10 μg/mL), or CEA (NeoBiotechnologies, 10 μg/mL) overnight at 4 °C.

Techniques: Immunohistochemistry, Staining, Microarray, Marker, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Control