monoclonal antibodies with the following specificities cd184 (cxcr4, clone 2b11)  (Thermo Fisher)

Journal: Cancer Cell International

Article Title: A1, an innovative fluorinated CXCR4 inhibitor, redefines the therapeutic landscape in colorectal cancer

doi: 10.1186/s12935-024-03584-y

Figure Lengend Snippet: Primer sequences and other details of the primers used in this study

Article Snippet: This mixture added 2.5 μL of the PE-conjugated anti-mouse CXCR4-specific antibody (R&D systems FAB21651P).

Techniques:

Journal: Cancer Cell International

Article Title: A1, an innovative fluorinated CXCR4 inhibitor, redefines the therapeutic landscape in colorectal cancer

doi: 10.1186/s12935-024-03584-y

Figure Lengend Snippet: CXCR4 expression in MEF and CT26 cell lines. ( A ) The bar graph illustrates the relative gene expression levels of CXCR4 in CT26 and MEF cell lines. ( B ) The bar graph represents the quantity of CXCR4-positive cells within CT26 and MEF cell populations, as quantified by flow cytometry. Below, the bar graph shows representative flow cytometry dot plots, showcasing the percentage of CXCR4-positive cells in each cell line (5.2% for MEF and 32% for CT26). The experiments were performed in triplicate. The results are expressed as the mean ± standard deviation (SD) from three independent experiments, with ** denoting statistical significance at P < 0.01 and **** indicating a statistical significance level of P < 0.0001

Article Snippet: This mixture added 2.5 μL of the PE-conjugated anti-mouse CXCR4-specific antibody (R&D systems FAB21651P).

Techniques: Expressing, Gene Expression, Flow Cytometry, Standard Deviation

Journal: Cancer Cell International

Article Title: A1, an innovative fluorinated CXCR4 inhibitor, redefines the therapeutic landscape in colorectal cancer

doi: 10.1186/s12935-024-03584-y

Figure Lengend Snippet: Comparative CXCR4, NFκB , and MMP-9 gene expression levels across CT-26 untreated and treated cells. The accompanying graph illustrates the relative gene expression levels in cells subjected to treatment with CXCL12 either alone or in combination with A1. The data is represented as Mean ± SD derived from three independent experiments. Significance levels are indicated with **** for P < 0.0001 and *** for P < 0.001, demonstrating the statistical differences among the treatment groups

Article Snippet: This mixture added 2.5 μL of the PE-conjugated anti-mouse CXCR4-specific antibody (R&D systems FAB21651P).

Techniques: Gene Expression, Derivative Assay

Journal: Cancer Cell International

Article Title: A1, an innovative fluorinated CXCR4 inhibitor, redefines the therapeutic landscape in colorectal cancer

doi: 10.1186/s12935-024-03584-y

Figure Lengend Snippet: Comparative analysis of the effect of 60 μg/ml A1 on the number of CXCR4 + CT-26 cells compared to treatment with 100 ng/ml CXCL12. This graph illustrates the alterations in the number of CXCR4 − expressing cells in response to a dosage of 60 μg/ml of drug A1 in comparison to cells treated with 100 ng/ml of CXCL12, which is a recognized chemokine ligand for CXCR4. The experiments were conducted in triplicate, and the data are presented as Mean ± SD. The treatment groups are evaluated to assess the relative impact of A1 on the CXCR4 + cell populations in contrast to the stimulation induced by CXCL12. Statistical significance is denoted by ****P < 0.0001, indicating a highly significant difference between the treated groups. The error bars represent the standard deviation among replicates, ensuring the experimental results' reproducibility

Article Snippet: This mixture added 2.5 μL of the PE-conjugated anti-mouse CXCR4-specific antibody (R&D systems FAB21651P).

Techniques: Expressing, Comparison, Standard Deviation

Journal: Cancer Cell International

Article Title: A1, an innovative fluorinated CXCR4 inhibitor, redefines the therapeutic landscape in colorectal cancer

doi: 10.1186/s12935-024-03584-y

Figure Lengend Snippet: CT-26 cell migration analysis after treatment with CXCL12, A1, and AMD3100. The graph illustrates the percentage of migrating CT-26 cells following exposure to 300 ng/ml CXCL12, 10 µm/ml A1, and 10 mM/ml AMD3100. Four experimental conditions are represented: ( a ) untreated cells (baseline migration), ( b ) cells treated with CXCL12 (control group to induce migration), ( c ) cells treated with AMD3100 (CXCR4 antagonist), and ( d ) cells treated with A1 (experimental drug). The results are expressed as mean ± SD from triplicate experiments. Statistical significance is indicated as follows: *P < 0.05, **P < 0.01, and ***P < 0.001, reflecting varying degrees of significance compared to the control group treated with CXCL12. These findings emphasize the effects of A1 and AMD3100 on the migratory behavior of CT-26 cells in response to chemotactic stimulation by CXCL12. Error bars denote the standard deviation between replicate experiments, ensuring the data's reliability

Article Snippet: This mixture added 2.5 μL of the PE-conjugated anti-mouse CXCR4-specific antibody (R&D systems FAB21651P).

Techniques: Migration, Control, Standard Deviation

Journal: Cancer Cell International

Article Title: A1, an innovative fluorinated CXCR4 inhibitor, redefines the therapeutic landscape in colorectal cancer

doi: 10.1186/s12935-024-03584-y

Figure Lengend Snippet: Comparative analysis of gene expression levels in tumor tissues of control, AMD3100, and A1-treated mice. The chart illustrates the relative expression levels of ( A ) CXCR4 , ( B ) FGF , ( C ) VEGF , ( D ) IL-10 , and ( E ) TGF-β genes within tumor tissues derived from three distinct groups: the control group (untreated), the AMD3100-treated group, and the A1-treated group of mice. Gene expression levels were quantified using qRT-PCR, and all experiments were performed in triplicate. The data is presented as Mean ± SEM, reflecting the variability among biological replicates. Statistical significance between the groups is denoted by *P < 0.05 and **P < 0.01, underscoring the differential expression of genes induced by AMD3100 and A1 treatments compared to the control group. These findings elucidate the modulatory effects of the treatments on critical genes implicated in tumor progression, angiogenesis, and immune regulation. Error bars reflect the standard error of the mean, thereby highlighting the consistency of the experimental outcomes

Article Snippet: This mixture added 2.5 μL of the PE-conjugated anti-mouse CXCR4-specific antibody (R&D systems FAB21651P).

Techniques: Gene Expression, Control, Expressing, Derivative Assay, Quantitative RT-PCR, Quantitative Proteomics

Journal: Cancer Cell International

Article Title: A1, an innovative fluorinated CXCR4 inhibitor, redefines the therapeutic landscape in colorectal cancer

doi: 10.1186/s12935-024-03584-y

Figure Lengend Snippet: The overall structure of CXCR4 (dimer) and the CXCR4 binding site (groove) for CXCL12, and the inhibitors binding pocket as well as a close view of the studied ligands interactions in the CXCR4 binding pocket; a . A1, b . ITD and c . AMD3100. Green dashes represent salt bridges, velvet dashes represent π- π electrostatic interactions, orange dashes represent π- cation and π- anion interactions, yellow dashes represent conventional hydrogen bonds, and light blue dashes represent halogen bonds

Article Snippet: This mixture added 2.5 μL of the PE-conjugated anti-mouse CXCR4-specific antibody (R&D systems FAB21651P).

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Fine Definition of the CXCR4-Binding Region on the V3 Loop of Feline Immunodeficiency Virus Surface Glycoprotein

doi: 10.1371/journal.pone.0010689

Figure Lengend Snippet: The detection of CXCR4 and CD134 was carried out by using CXCR4 or CD134-specific antibody, followed by anti-mouse (CXCR4) or anti-rabbit (CD134) IgG conjugated with phycoerythrin. Binding of FIV SU-Fc to cells was measured by using a phycoerythrin-conjugated anti-Fc antibody, in the presence and absence of the CXCR4 antagonist AMD3100. Cells were incubated with SU-Fc or pre-treated with AMD3100 for 30 min prior to the addition of SU-Fc. Results are representative of three independent determinations.

Article Snippet: Human specific anti-CXCR4 antibody 12G5 – was obtained from BD Biosciences (Franklin Lakes, NJ), feline anti-CXCR4 monoclonal antibody was purchased from R&D Systems, Inc (Minneapolis, MN), which is highly specific for feline CXCR4.

Techniques: Binding Assay, Incubation

Journal: PLoS ONE

Article Title: Fine Definition of the CXCR4-Binding Region on the V3 Loop of Feline Immunodeficiency Virus Surface Glycoprotein

doi: 10.1371/journal.pone.0010689

Figure Lengend Snippet: ( A ) . Sequences and amino acid percent frequency of the “N44 region” of FIV isolates. The number next to each residue indicates the frequency at which the corresponding residue is found among over 200 FIV envelope sequences published in Genbank. ( B ) Expression of FIV SU-Fc with amino acid substitutions. FIV SU-Fc mutants were generated by site-directed mutagenesis, expressed by stable transfection of CHO-K1 cells and batch purified from cell supernatants by affinity chromatography over protein A-Sepharose. 100 ng of SU-Fc (wt and mutants) were subjected to SDS-PAGE under reducing conditions and reacted with HRP-conjugated anti-human IgG1antibody. The labels are located above in which amino acid substitutions have been introduced, and the resulting mutants were assessed for abilities to bind CXCR4 and CD134.

Article Snippet: Human specific anti-CXCR4 antibody 12G5 – was obtained from BD Biosciences (Franklin Lakes, NJ), feline anti-CXCR4 monoclonal antibody was purchased from R&D Systems, Inc (Minneapolis, MN), which is highly specific for feline CXCR4.

Techniques: Expressing, Generated, Mutagenesis, Stable Transfection, Purification, Affinity Chromatography, SDS Page

Journal: PLoS ONE

Article Title: Fine Definition of the CXCR4-Binding Region on the V3 Loop of Feline Immunodeficiency Virus Surface Glycoprotein

doi: 10.1371/journal.pone.0010689

Figure Lengend Snippet: Effects of Amino Acid Substitutions in SU on CXCR4-binding Ability in 3201 Cells.

Article Snippet: Human specific anti-CXCR4 antibody 12G5 – was obtained from BD Biosciences (Franklin Lakes, NJ), feline anti-CXCR4 monoclonal antibody was purchased from R&D Systems, Inc (Minneapolis, MN), which is highly specific for feline CXCR4.

Techniques:

Journal: PLoS ONE

Article Title: Fine Definition of the CXCR4-Binding Region on the V3 Loop of Feline Immunodeficiency Virus Surface Glycoprotein

doi: 10.1371/journal.pone.0010689

Figure Lengend Snippet: Effects of Amino Acid Substitutions in SU on CXCR4-binding Ability in SupT1 Cells.

Article Snippet: Human specific anti-CXCR4 antibody 12G5 – was obtained from BD Biosciences (Franklin Lakes, NJ), feline anti-CXCR4 monoclonal antibody was purchased from R&D Systems, Inc (Minneapolis, MN), which is highly specific for feline CXCR4.

Techniques:

Journal: Nature Communications

Article Title: Computationally designed GPCR quaternary structures bias signaling pathway activation

doi: 10.1038/s41467-022-34382-7

Figure Lengend Snippet: a Mutations designed to selectively stabilize the CXCR4 open-dimer conformation without affecting CXCR4 monomer stability were identified in the extracellular and TMH regions. b Mutations designed to selectively stabilize the CXCR4 closed-dimer conformation without affecting CXCR4 monomer stability were identified in the extracellular region. Key atomic contacts are represented as red dotted lines. c Schematic conformational energy landscapes of CXCR4 dimerization in the inactive and active states for the open-dimer stabilizing designs. d Schematic conformational energy landscapes of CXCR4 dimerization in the inactive and active states for the closed-dimer stabilizing designs. c , d The dimerization energies reported in Supplementary Table were used to plot the energy landscapes. The monomer energies and energy barriers between states are fictitious and were not predicted by our simulations. e Ranking of the CXCR4 variants based on changes in buried surface area upon dimerization (ΔSASA) calculated from the predicted models in the active state. The ΔSASA is reported for the most occupied dimer conformation for each variant: L194R open-dimer state, WT open-dimer state, N192W closed-dimer state, W195L closed-dimer state. Larger buried ΔSASA are predicted to correlate with enhanced dimerization propensity (see Supplementary Table ).

Article Snippet: Endogenous CXCR4 expression on the surface of HEK and U87 cells was monitored by flow cytometry using CXCR4-specific phycoerythrin-conjugated mAb 12G5 1:20 or the corresponding isotype control (R&D Systems) in a BD FACS LSR Fortessa cytometer (BD Biosciences).

Techniques: Variant Assay

Journal: Nature Communications

Article Title: Computationally designed GPCR quaternary structures bias signaling pathway activation

doi: 10.1038/s41467-022-34382-7

Figure Lengend Snippet: a (Left) Schematic representation of the CXCR4 dimerization BRET-based assay. (Right) CXCR4 association was measured by BRET before (black) and after agonist stimulation (gray) in HEK293T cells transfected with CXCR4-RLuc and its counterpart CXCR4-YFP, WT, or mutant as indicated. BRET 480-YFP was measured after the addition of coel-h (10 min) and CXCL12 (15 min). Data shown represent the mean ± SEM of three independent experiments and are expressed as net BRET (calculated by subtracting background luminescence). Statistical significance was assessed using a two-way ANOVA followed by a Šídák’s multiple comparisons test: # p = 0.007, ## p < 0.0001, n.s. not significant p > 0.05 are used to compare BRET values between basal to CXCL12-treated conditions and † p = 0.0004, †† p < 0.0001 are used to compare basal BRET values between the mutants. b (Left) Schematic representation of the BRET-based ligand-induced Gi activation assay. (Right) CXCL12-promoted Gi activation measured by BRET in HEK293T cells transfected with HA-CXCR4, WT or mutant as indicated, Gαi1-RLucII, Gβ1, and Gγ2-GFP10. BRET 400-GFP10 was measured after the addition of coel-400a (10 min) and CXCL12 (3 min). c (Left) Schematic representation of the BRET-based EPAC sensor to measure cAMP production. (Right) CXCL12-promoted EPAC inhibition was measured by BRET in HEK293T cells transfected with HA-CXCR4, WT or mutant as indicated, and RLuc-EPAC-YFP. BRET 480-YFP , reporting the conformation rearrangement of the EPAC sensor from an open to a closed conformation, was measured after the addition of coel-h (10 min) and CXCL12 (5 min). b , c CXCR4 mutations predicted to stabilize the open-dimer or the closed-dimer conformation are annotated with a blue or red dimer symbol, respectively. Data shown represent the mean ± SEM of at least three independent experiments and are expressed as ΔBRET (agonist-promoted BRET).

Article Snippet: Endogenous CXCR4 expression on the surface of HEK and U87 cells was monitored by flow cytometry using CXCR4-specific phycoerythrin-conjugated mAb 12G5 1:20 or the corresponding isotype control (R&D Systems) in a BD FACS LSR Fortessa cytometer (BD Biosciences).

Techniques: Bioluminescence Resonance Energy Transfer, Transfection, Mutagenesis, Activation Assay, Inhibition

Journal: Nature Communications

Article Title: Computationally designed GPCR quaternary structures bias signaling pathway activation

doi: 10.1038/s41467-022-34382-7

Figure Lengend Snippet: a (Left) Schematic representation of the BRET-based ligand-induced β-arrestin-2 (βarr2) translocation assay. (Right) CXCL12-promoted βarr2 recruitment to CXCR4 measured by BRET in HEK293T cells transfected with CXCR4-RLuc, WT or mutant as indicated, and βarr2-YFP. BRET 480-YFP between CXCR4-RLuc and βarr2-YFP was measured after the addition of coel-h (10 min) and CXCL12 (15 min). Data shown represent the mean ± SEM of at least three independent experiments and are represented as ΔBRET. b (Left) Phosphorylation at S324/5 of WT and W195 5.34 L CXCR4 promoted by stimulation with 200 nM CXCL12 for 30 min detected using an anti pS324/5 antibody (pCXCR4 indicates the CXCR4-S324/5 phosphorylation band; non-SP correspond to a non-specific band). (Right) Quantification of phosphorylation bands normalized as a function of the intensity of the total HA-CXCR4 detected using an anti-HA antibody. Shown in the inset is the fold increase in phosphorylation over basal levels. Data shown represent the mean ± SEM of three independent experiments. Statistical significance was assessed using unpaired t test. # p = 0.001, ## p < 0.0001, n.s. not significant p > 0.05. c (Left) Schematic representation of ERK activation by CXCR4. (Right) ERK phosphorylation in U87 stably expressing equivalent levels of WT and W195 5.34 L CXCR4 induced by stimulation with 10 nM CXCL12 for the indicated times was monitored by HTRF. CXCR4 mutations predicted to stabilize the open-dimer or the closed-dimer conformation are annotated with a blue or red dimer symbol, respectively. Data shown represent the mean ± SEM of at least three independent experiments.

Article Snippet: Endogenous CXCR4 expression on the surface of HEK and U87 cells was monitored by flow cytometry using CXCR4-specific phycoerythrin-conjugated mAb 12G5 1:20 or the corresponding isotype control (R&D Systems) in a BD FACS LSR Fortessa cytometer (BD Biosciences).

Techniques: Translocation Assay, Transfection, Mutagenesis, Phospho-proteomics, Activation Assay, Stable Transfection, Expressing

Journal: Nature Communications

Article Title: Computationally designed GPCR quaternary structures bias signaling pathway activation

doi: 10.1038/s41467-022-34382-7

Figure Lengend Snippet: a Surface representation of the CXCR4 inactive state monomeric structure highlighting the distinct oligomerization interfaces controlling either β-arrestin recruitment (extracellular side and TM core of TMH5, blue) or Gi activation and nanocluster formation (intracellular side of TMH6, orange). b The hotspot binding sites controlling CXCR4 oligomerization through TMH5 (designed residues in red) are poorly conserved in the chemokine receptor family, except for the β-arrestin signaling switch W5.34. Aromatic residues are highly enriched at position 5.34 of other dimerizing GPCR families. c Conserved position and conformation of W5.34 in human chemokine receptor X-ray structures. The superposition of W5.34 conformations is shown in the center. d Conservation of the β-arrestin signaling switch in the TM5 of vasopressin/oxytocin receptors and opioid receptors.

Article Snippet: Endogenous CXCR4 expression on the surface of HEK and U87 cells was monitored by flow cytometry using CXCR4-specific phycoerythrin-conjugated mAb 12G5 1:20 or the corresponding isotype control (R&D Systems) in a BD FACS LSR Fortessa cytometer (BD Biosciences).

Techniques: Activation Assay, Binding Assay

Journal: Cell Reports Medicine

Article Title: Targeted blockade of immune mechanisms inhibit B precursor acute lymphoblastic leukemia cell invasion of the central nervous system

doi: 10.1016/j.xcrm.2021.100470

Figure Lengend Snippet: B-ALL transit through the skull and vertebral bones in an MLL -rearranged B-ALL Human MLLr NSG-passaged leukemic blasts were injected into the right femurs of NSG mice and treated with IgG1-Fc control or AMD3100 or rOPG-Fc or combined AMD3100 and rOPG-Fc (n = 3 mice per group). After 4 weeks of leukemic blast injection and treatment, mice were euthanized. The whole brain with an intact skull and vertebral bones with a spinal cord were fixed, sectioned, and stained with anti-human CD19 or CXCR4 antibodies. (A–D) Representative sagittal sections of the brain and skull showing calvarial bone marrow and brain subarachnoid space in the (1) caudal region, and vertebra showing the bone marrow, subarachnoid space, spinal cord, and skeletal muscles of IgG1-Fc- or AMD3100- or rOPG-Fc- or combined AMD3100- and rOPG-Fc-treated mice. Brown staining indicates CD19 or CXCR4 abundance. The box represents a magnified view. The dotted outline indicates leukemic blasts. The red arrows indicate bone channels that connect the bone marrow and subarachnoid space. (E–I) CD19 protein (y axis) was quantified in multiple ROIs (25 μm) from the (E) calvarial and (F) vertebral bone marrow, (G) and brain and (H) spinal cord subarachnoid space, and (I) skeletal muscles of the IgG1-Fc-, AMD3100-, rOPG-Fc-, or AMD3100 + rOPG-Fc-treated mice (x axis). The graph shows the mean staining intensity (2 μm per pixel) of CD19 protein (au). Each dot represents multiple ROIs from 3 biological replicates and 2 or 3 technical replicates (n = 3 mice per group). Data are means ± SDs. All of the data were analyzed using non-parametric Kruskal-Wallis 1-way analysis of variance (ANOVA) (95% confidence interval) among the 3 groups; p values represent post hoc Dunn’s test, in which ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. Scale bars, 100 μm. See also , , and .

Article Snippet: Moreover, single-cell suspensions isolated from the bone marrow of BCR-ABL1 PDX mice that were treated with IgG1-Fc control were stained with human-specific cell-surface antibodies for CXCR4-PECy7 (1:150 dilution, clone: 12G5, BD Biosciences), CD45-FITC (1:30 dilution, clone: 2D1, BD Biosciences), CD19-PE (1:30 dilution, clone: 4G7, BD Biosciences).

Techniques: Injection, Staining

Journal: Cell Reports Medicine

Article Title: Targeted blockade of immune mechanisms inhibit B precursor acute lymphoblastic leukemia cell invasion of the central nervous system

doi: 10.1016/j.xcrm.2021.100470

Figure Lengend Snippet: B-ALL blast transit by breaching the blood-cerebrospinal fluid barrier in MLL -rearranged PDX mice Human MLLr NSG-passaged leukemic blasts were injected into the right femurs of NSG mice and treated with IgG1-Fc control or AMD3100 or rOPG-Fc or combined AMD3100 and rOPG-Fc (n = 4 mice per group). After 6 weeks of leukemic blast injection and treatment, the mice were euthanized. The whole brain with an intact skull and vertebral bones spinal cord was fixed, sectioned, and stained with anti-human CD19 or CXCR4 antibodies. (A–D) Representative sagittal sections of the brain and skull showing calvarial bone marrow and brain subarachnoid space in the (1) caudal region, lateral ventricle (LV) and fourth ventricles (FVs), and vertebra showing the bone marrow, subarachnoid space, spinal cord, and skeletal muscles of IgG1-Fc- or AMD3100- or rOPG-Fc- or combined AMD3100- and rOPG-Fc-treated mice. Brown staining indicates CD19 or CXCR4 abundance. The box represents a magnified view. The dotted outline indicates leukemic blasts. The red arrows indicate bone channels that connect the bone marrow and subarachnoid space. (E–J) CD19 protein (y axis) was quantified in multiple ROIs (25 μm) from the (E) calvarial and (F) vertebral bone marrow, (G) brain and (H) spinal cord subarachnoid space, (I) skeletal muscles, and (J) lateral and fourth brain ventricles of the IgG1-Fc-treated, AMD3100-treated, rOPG-Fc-treated, or AMD3100 + rOPG-Fc-treated mice (x axis). The graph shows the mean staining intensity (2 μm per pixel) of CD19 protein (au). Each dot represents multiple ROIs from 4 biological replicates and 2 or 3 technical replicates (n = 4 mice per group). Data are means ± SDs. All of the data were analyzed using non-parametric Kruskal-Wallis 1-way ANOVA (95% confidence interval) among the 3 groups; p values represent post hoc Dunn’s test, in which ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. Scale bars, 100 μm. See also , , and .

Article Snippet: Moreover, single-cell suspensions isolated from the bone marrow of BCR-ABL1 PDX mice that were treated with IgG1-Fc control were stained with human-specific cell-surface antibodies for CXCR4-PECy7 (1:150 dilution, clone: 12G5, BD Biosciences), CD45-FITC (1:30 dilution, clone: 2D1, BD Biosciences), CD19-PE (1:30 dilution, clone: 4G7, BD Biosciences).

Techniques: Injection, Staining

Journal: Cell Reports Medicine

Article Title: Targeted blockade of immune mechanisms inhibit B precursor acute lymphoblastic leukemia cell invasion of the central nervous system

doi: 10.1016/j.xcrm.2021.100470

Figure Lengend Snippet: Schematic representation of B-ALL blast transit routes to the CNS Sagittal sections of the mouse brain showing RANKL- and CXCR4-mediated B-ALL transit routes. (A) In the PDX model, we show that B-ALL blasts fail to breach the blood-brain barrier, as we did not observe leukemic blasts in the brain parenchyma. However, at a late disease stage, BCR-ABL1 and MLLr B-ALL blasts may have reached the subarachnoid space by breaching the blood-leptomeningeal barrier. (B) We identified a calvarial or vertebral bone marrow-mediated leukemic blast transit into the subarachnoid space in PDX mice. RANKL antagonist rOPG-Fc treatment protected the skull and vertebral bone marrow from B-ALL blast invasion and subsequent skull or vertebral bone marrow-mediated leukemic blast transit into the subarachnoid space. (C) BCR-ABL1 B-ALL blasts did not breach the blood-cerebrospinal fluid barrier in PDX mice. In contrast, CD19 + CXCR4 + MLLr B-ALL blasts breached the blood-cerebrospinal fluid barrier found in the LV, third ventricle, and FV of the brain at a late-disease stage, and CXCR4 and rOPG-Fc antagonism using AMD3100 and rOPG-Fc prevented B-ALL blast transit through the blood-cerebrospinal fluid barrier.

Article Snippet: Moreover, single-cell suspensions isolated from the bone marrow of BCR-ABL1 PDX mice that were treated with IgG1-Fc control were stained with human-specific cell-surface antibodies for CXCR4-PECy7 (1:150 dilution, clone: 12G5, BD Biosciences), CD45-FITC (1:30 dilution, clone: 2D1, BD Biosciences), CD19-PE (1:30 dilution, clone: 4G7, BD Biosciences).

Techniques:

Journal: Cell Reports Medicine

Article Title: Targeted blockade of immune mechanisms inhibit B precursor acute lymphoblastic leukemia cell invasion of the central nervous system

doi: 10.1016/j.xcrm.2021.100470

Figure Lengend Snippet:

Article Snippet: Moreover, single-cell suspensions isolated from the bone marrow of BCR-ABL1 PDX mice that were treated with IgG1-Fc control were stained with human-specific cell-surface antibodies for CXCR4-PECy7 (1:150 dilution, clone: 12G5, BD Biosciences), CD45-FITC (1:30 dilution, clone: 2D1, BD Biosciences), CD19-PE (1:30 dilution, clone: 4G7, BD Biosciences).

Techniques: Recombinant, Produced, Plasmid Preparation, Construct, Software