Journal: Journal of Translational Medicine

Article Title: Myeloid leukemia-derived galectin-1 downregulates CAR expression to hinder cytotoxicity of CAR T cells

doi: 10.1186/s12967-023-04832-x

Figure Lengend Snippet: Galectins are highly expressed in myeloid leukemia cells A , Heat maps showing suppressive genes differentially expressed between myeloid and lymphoid leukemia cell lines and between primary AML and ALL samples. Genes are those that are up or down-regulated more than two-fold, except galectin-9 in primary AML and ALL samples. AML and ALL primary samples had two biological replicates, respectively. B–E , Histograms showing the expression of galectin-1, galectin-3, galectin-12, and galectin-9 in AML and ALL cell lines; Histograms and the MFI of galectin-1, galectin-3,and galectin-12 in primary AML and ALL samples. Data shown are mean ± SEM of three experimental replicates

Article Snippet: We added NH 4 Cl (20 mM), galectin-1 specific inhibitor anginex (Jiangsu Jitai, 10 µM), galectin-3 specific inhibitor G3-C12 (MCE, 30 µM), recombinant galectin-1 protein (Acro, 50 µg/ml), recombinant galectin 3 protein (MCE, 20 µg/ml), recombinant galectin 12 protein (MCE, 10 µg/ml) and galectin 12 antibody (CUSABIO) to the coculture system.

Techniques: Expressing

Journal: Journal of Translational Medicine

Article Title: Myeloid leukemia-derived galectin-1 downregulates CAR expression to hinder cytotoxicity of CAR T cells

doi: 10.1186/s12967-023-04832-x

Figure Lengend Snippet: Myeloid leukemia cells use galectin-1 to block CAR T cells function A , The specific lysis after coculture with U937 CD33 cells at a 1:5 in the presence of recombinant galectin 3 protein (20 µg/ml) and galectin-3 specific inhibitor G3-C12 (30 µM). B , The specific lysis after coculture with U937 CD33 cells at a 1:1 in the presence of recombinant galectin 12 protein (10 µg/ml) and galectin 12 antibody. C , The specific lysis, the percentage of CD69 expression in CD33 CAR T cells, and the percentage of surface CAR expression in CD33 CAR T cells, after coculture with U937 CD33 cells at a 1:10 E:T ratio for 12 h, in the presence of 10 µM galectin-1 inhibitory peptide (ANGINEX). D , The specific lysis and the percentage of surface CAR expression in CD33 CAR T cells, after coculture with U937 CD33 cells at a 50 µg/ml recombinant galectin-1 protein. E , The specific lysis and the percentage of surface CAR expression in CD33 CAR T cells, after coculture with U937 CD33 cells at a 1:1 ratio for 12 h in the presence of 10 µM galectin-1 inhibitory peptide (ANGINEX). F, Total number of CD123, CD33,CD19 and CLL-1 CAR-T cells in the experimental group with 50 µg/ml recombinant galectin-1 protein and in the control group. G , Annexin V positivity in CD123, CD33, CD19 and CLL1 CAR-T cells from experimental and control groups using 50 µg/ml recombinant galectin-1 protein

Article Snippet: We added NH 4 Cl (20 mM), galectin-1 specific inhibitor anginex (Jiangsu Jitai, 10 µM), galectin-3 specific inhibitor G3-C12 (MCE, 30 µM), recombinant galectin-1 protein (Acro, 50 µg/ml), recombinant galectin 3 protein (MCE, 20 µg/ml), recombinant galectin 12 protein (MCE, 10 µg/ml) and galectin 12 antibody (CUSABIO) to the coculture system.

Techniques: Blocking Assay, Lysis, Recombinant, Expressing

Journal: Cancers

Article Title: Autocrine Signaling of NRP1 Ligand Galectin-1 Elicits Resistance to BRAF-Targeted Therapy in Melanoma Cells

doi: 10.3390/cancers12082218

Figure Lengend Snippet: Galectin-1 upregulation in melanoma samples and cell lines resistant to BRAF-targeted therapy. ( A ) The graph shows Galectin-1 (LGALS1 gene) mRNA expression variation in paired melanoma samples (obtained from seven patients, indicated by black diamonds) before treatment with BRAF inhibitors and after the onset of secondary resistance (Log2 ratio resistant/pretherapy), as well as in A375 and SK-MEL-28 melanoma cells before and after developing resistance to 2μM PLX-4720 (Log2 ratio Resistant/Parental; n = 3). ( B ) Analysis of Galectin-1 protein levels (by PikoKine ELISA assay) in conditioned media (CM) derived from either parental or PLX4720-resistant A375 and SK-MEL-28 (PLX-resistant) cells. Data are normalized to levels detected in the respective parental cells and represent the average of a technical duplicate. Statistical significance was assessed by t -test, comparing parental and their derived PLX-resistant cell lines; *** p < 0.0001.

Article Snippet: Galectin-1 concentrations in CM were measured by PikoKine ELISA assay (category EK0762; Boster Biological Technology, Pleasanton, CA, USA), according to the manufacturer’s protocol.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay