Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Development of SGN-CD47M: Protease-Activated Antibody Technology Enables Selective Tumor Targeting of the Innate Immune Checkpoint Receptor CD47

doi: 10.1158/1535-7163.MCT-24-0371

Figure Lengend Snippet: A protease-activated antibody approach using coiled-coil masking peptides improves the PK and tolerability of an antibody directed to mouse CD47. A, Masking impaired the binding of mIAP301 to mouse CD47 on HT-2 cells but could be restored upon cleavage with recombinant MMP. Symbols represent duplicate samples shown as mean ± SD. A, Decreased binding resulted in impaired RBC phagocytosis ( B ) in vitro . Representative data from three independent experiments. Symbols represent the mean ± SD from three biological replicates. *, P < 0.05; ***, P < 0.001 (one-way ANOVA followed by the Dunnett multiple comparisons test) compared with mIAP301. C, PK profiles of unmasked mIAP301 and Coil-MMP-mIAP301 in naïve BALB/c mice, which demonstrates strikingly different plasma exposure profiles because of avoidance of the large normal tissue sink for CD47. The dotted line denotes the level of quantitation in the experiment. It should be noted that the concentration of mIAP301 was only at or above the level of quantitation at the 15-minute time point. Symbols indicate n = 3 replicates and are shown as the mean ± SD. LOQ, level of quantitation. D , Antitumor activity of masked and unmasked mIAP301 antibodies was assessed in an A20 lymphoma model in BALB/c mice and showed similar antitumor activity (every two days for 8 cycles i.p. dosing). Error bars represent mean ± SD ( n = 6 mice per group). Statistical assessment for pairwise comparisons of tumor growth inhibition between treatment arms is provided in Supplementary Fig. S3. In tumor-bearing mice, masking decreased RBC depletion ( E ), and a comparison of bound antibody on RBCs vs. tumor cells provides evidence for tumor-specific mask activation ( F ). Data shown in E and F , are representative of two individual experiments. Symbols represent three biological replicates and are shown as the mean ± SD. *, P < 0.05; ***, P < 0.01; ***, P < 0.001 (one-way ANOVA followed by the Tukey multiple comparisons test) compared with mIAP301.

Article Snippet: Saturation binding ELISA experiments were performed using soluble recombinant human, mouse, or cyno CD47 (ACROBiosystems), which was diluted to an appropriate concentration in 50 mmol/L carbonate buffer at pH 9.6.

Techniques: Binding Assay, Recombinant, In Vitro, Clinical Proteomics, Quantitation Assay, Concentration Assay, Activity Assay, Inhibition, Comparison, Activation Assay

Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Development of SGN-CD47M: Protease-Activated Antibody Technology Enables Selective Tumor Targeting of the Innate Immune Checkpoint Receptor CD47

doi: 10.1158/1535-7163.MCT-24-0371

Figure Lengend Snippet: SGN-CD47M has impaired binding and in vitro activity that can be restored upon mask cleavage with MMPs. Unmasked anti-CD47 antibody hB6H12.3 shows binding to SW780 pancreatic tumor cells ( A ) and human RBCs ( B ), whereas SGN-CD47M shows negligible binding. Representative data from three independent experiments. Symbols represent duplicate samples shown as mean ± SD. SGN-CD47M that had been pretreated with MMPs is able to bind at levels comparable with hB6H12.3. The ability of hB6H12.3, SGN-CD47M, and MMP-cleaved SGN-CD47M to elicit macrophage-mediated phagocytosis against Jurkat T-cell lymphoma cells and human RBCs was evaluated and demonstrated that masking can impair in vitro function that can be largely restored upon mask cleavage. Representative data from two independent experiments. Symbols indicate n = 6 biological replicates and are represented as the mean ± SD.

Article Snippet: Saturation binding ELISA experiments were performed using soluble recombinant human, mouse, or cyno CD47 (ACROBiosystems), which was diluted to an appropriate concentration in 50 mmol/L carbonate buffer at pH 9.6.

Techniques: Binding Assay, In Vitro, Activity Assay

Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Development of SGN-CD47M: Protease-Activated Antibody Technology Enables Selective Tumor Targeting of the Innate Immune Checkpoint Receptor CD47

doi: 10.1158/1535-7163.MCT-24-0371

Figure Lengend Snippet: Intratumoral mask cleavage of SGN-CD47M leads to tumor cell binding and induction of tumor cell apoptosis in HT1080 xenograft tumors. A, SGN-CD47M mask cleavage was detected in plasma, spleen, and tumors of HT1080 tumor-bearing mice using Western blot analysis. Time-dependent preferential mask cleavage was observed in the tumor compared with plasma and spleen. Data are representative of three independent experiments. Intratumoral mask cleavage leads to tumor cell binding ( B ), as assessed by flow cytometry analysis of bound human antibody on dissociated tumor cells. C, Subsequent staining of a noncompeting anti-CD47 antibody demonstrates that hB6H12.3 and activated SGN-CD47M binding leads to a decrease in available CD47 receptors. D, Engagement of CD47 shows a trend for increased tumor cell apoptosis for hB6H12.3 and SGN-CD47M based on annexin V staining of dissociated tumor cells by flow cytometry. For B–D , the data are presented from a single independent experiment. For all panels, each experimental arm consisted of three biological replicates from individual mice and data presented are mean ± SEM. *, P < 0.05; ***, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (one-way ANOVA followed by the Tukey multiple comparisons test) compared with hB6H12.3. MFI, mean fluorescent index.

Article Snippet: Saturation binding ELISA experiments were performed using soluble recombinant human, mouse, or cyno CD47 (ACROBiosystems), which was diluted to an appropriate concentration in 50 mmol/L carbonate buffer at pH 9.6.

Techniques: Binding Assay, Clinical Proteomics, Western Blot, Flow Cytometry, Staining

Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Development of SGN-CD47M: Protease-Activated Antibody Technology Enables Selective Tumor Targeting of the Innate Immune Checkpoint Receptor CD47

doi: 10.1158/1535-7163.MCT-24-0371

Figure Lengend Snippet: SGN-CD47M shows improved tolerability in nonhuman primates because of decreased systemic CD47 engagement. A, The extent of decrease in red cell mass (measured by hematocrit) was similar to animals receiving 20 mg/kg SGN-CD47M or 1 mg/kg hB6H12.3. Treatment at these doses resulted in about 65% decrease in red cell mass at the nadir that trended toward recovery by study day 22, and a dose of 10 mg/kg of SGN-CD47M resulted in about 44% decrease in red cell mass. Assessment of antibody concentrations in plasma ( B ) and blood pellets ( C ) provides supporting data indicating that SGN-CD47M remains masked in vivo . D, The production of cytokines such as MCP-1 for SGN-CD47M and hB6H12.3 was similar despite a 20-fold higher dose. For all panels, symbols represent two biological replicates from individual animals, shown as the mean ± SD. MCP-1, monocyte chemoattractant protein 1.

Article Snippet: Saturation binding ELISA experiments were performed using soluble recombinant human, mouse, or cyno CD47 (ACROBiosystems), which was diluted to an appropriate concentration in 50 mmol/L carbonate buffer at pH 9.6.

Techniques: Clinical Proteomics, In Vivo

Journal: Molecular Cancer Therapeutics

Article Title: Preclinical Development of SGN-CD47M: Protease-Activated Antibody Technology Enables Selective Tumor Targeting of the Innate Immune Checkpoint Receptor CD47

doi: 10.1158/1535-7163.MCT-24-0371

Figure Lengend Snippet: SGN-CD47M is stable in human cancer whole blood and plasma. Binding of FITC-labeled hB6H12.3 and SGN-CD47M to whole blood of human patients with cancer was assessed by flow cytometry after 20 hours incubation at 37°C ( A ). SGN-CD47M stability in plasma samples of human patients with cancer was assessed using a CD47-binding ELISA ( B ) after 96 hours of incubation at 37°C. At the concentration of antibody used in the experiment (20 μg/mL), SGN-CD47M does not bind to recombinant CD47 and is not detected by ELISA. Therefore, binding indicates that the masking domain of SGN-CD47M has been proteolyzed. Under these conditions, no binding of SGN-CD47M spiked into any of the 19 samples was detected, whereas spiked control samples of hB6H12.3 yielded quantitative detection of 20 μg/mL. All samples spiked with SGN-CD47M were quantified below 350 ng/mL, which equates to <2% binding of hB6H12.3. Symbols represent two biological replicates for each donor, shown as the mean ± SD. CRC, colorectal cancer; MFI, mean fluorescent index; NSCLC, non–small cell lung cancer.

Article Snippet: Saturation binding ELISA experiments were performed using soluble recombinant human, mouse, or cyno CD47 (ACROBiosystems), which was diluted to an appropriate concentration in 50 mmol/L carbonate buffer at pH 9.6.

Techniques: Clinical Proteomics, Binding Assay, Labeling, Flow Cytometry, Incubation, Enzyme-linked Immunosorbent Assay, Concentration Assay, Recombinant, Control

Journal: Frontiers in Immunology

Article Title: SIRPγ-CD47 Interaction Positively Regulates the Activation of Human T Cells in Situation of Chronic Stimulation

doi: 10.3389/fimmu.2021.732530

Figure Lengend Snippet: Key resource table.

Article Snippet: Recombinant protein , CD47 , AcroBiosystems, Newark, DE, USA , CD7-H82F6 , Human , .

Techniques: Produced, Recombinant, Isolation, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: SIRPγ-CD47 Interaction Positively Regulates the Activation of Human T Cells in Situation of Chronic Stimulation

doi: 10.3389/fimmu.2021.732530

Figure Lengend Snippet: The KWAR23 is a Pan SIRP mAb which impairs SIRPα and SIRPγ interactions with CD47. (A) Human monocytes and T cells were labeled with KWAR23, anti-SIRPα mAb (18D5) or an isotype control (ISO) all at 10,000 ng/ml and further analyzed by FACS. (B) The percentage of stained cells (monocytes and T cells) is presented regarding the concentration of mAb ( n = 1). (C) KWAR23-binding specificity towards SIRPγ was evaluated by FACS staining on wild-type (WT) and SIRPγ−KO Jurkat cells (upper overlay). LSB2.20 (anti-SIRPγ)KWAR23 recognized staining was used as control (lower overlay). (D) Specificity of mAb used in (A) and (B) were assessed by ELISA against recombinant SIRPα and SIRPγ proteins. Binding activity is presented as normalized absorbance (DO) at 450 nm in function of concentration of mAb (ng/ml), n = 2–4 individual experiments (mean ± SEM). (E) Antagonist activity of the KWAR23, the anti-SIRPα (18D5), and the anti-SIRPγ (LSB2.20) mAbs to SIRPα/CD47 (left panel) and SIRPγ/CD47 (right panel) binding was measured by ELISA and presented as normalized absorbance (DO) at 450 nm in function of mAb added (ng/ml). n = 3 individual experiments (mean ± SEM).

Article Snippet: Recombinant protein , CD47 , AcroBiosystems, Newark, DE, USA , CD7-H82F6 , Human , .

Techniques: Labeling, Control, Staining, Concentration Assay, Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Activity Assay

Journal: Frontiers in Immunology

Article Title: SIRPγ-CD47 Interaction Positively Regulates the Activation of Human T Cells in Situation of Chronic Stimulation

doi: 10.3389/fimmu.2021.732530

Figure Lengend Snippet: SIRPγ expression varies on T-cell subpopulations, upon T-cell activation and is not implicated in Tregs function. (A) T-cell subpopulations gating strategy and their SIRPγ and CD47 expressions. (B) Geometric mean of SIRPγ and (C) CD47 on T-cell subpopulations; each symbol represents independent HV. Statistical analysis was performed by a Friedman test followed by a Dunn’s multiple comparison test. (D – F) SIRPγ and CD47 expressions were analyzed after T-cell stimulation with coated anti-CD3 and anti-CD28 mAbs. (D) Overlay allowing the comparison of SIRPγ (left) and CD47 (right) expression on unstimulated purified T cells (i.e., Day 0 in blue) vs . proliferating CPD low T cells (i.e., Day 5 in red). (E) Geometric mean (± SEM) of SIRPγ or (F) CD47 on T cells as a function of their number of divisions, n = 10 HV analyzed in two independent experiments. Kruskal-Wallis test was used. (G) Treg gating strategy based on CD25 and CD127 expression is represented as dot plots; representative histograms of SIRPγ and CD47 staining on Tregs subpopulations are shown. (H) Geometric mean of SIRPγ or (I) CD47 on Tregs are plotted. Each symbol represents independent HV. Statistical analysis was performed by a Friedman test followed by a Dunn’s multiple comparisons test. (J) A representative histogram of T effector (Teff) and T regulatory (Treg) cell proliferation under control conditions is shown at Day 5. (K) The effect of SIRPγ blockade with KWAR23 (10 µg/ml) was evaluated on T-cell proliferation upon OKT3 and anti-CD28 stimulation (both at 1 µg/ml). n = 2 independent experiments. (L) The consequences of SIRPγ blockade on Treg suppressive function was assessed by coculturing ef450 CPD-labeled Teff with ef670 CPD-labeled Treg cells in the presence or not of stimulatory molecule (OKT3) with or without SIRPγ blockade (KWAR23). Percentage of CPD low effector T cells (± SEM) is presented regarding culture conditions. n = 4 independent experiments. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001; ns, not significant.

Article Snippet: Recombinant protein , CD47 , AcroBiosystems, Newark, DE, USA , CD7-H82F6 , Human , .

Techniques: Expressing, Activation Assay, Comparison, Cell Stimulation, Purification, Staining, Control, Labeling

Journal: Frontiers in Immunology

Article Title: SIRPγ-CD47 Interaction Positively Regulates the Activation of Human T Cells in Situation of Chronic Stimulation

doi: 10.3389/fimmu.2021.732530

Figure Lengend Snippet: CD47 and SIRPγ do not colocalize on T cells. (A, B) Confocal microscopy imaging showing SIRPγ (red) and CD47 (green) cell surface distribution on Jurkat T (A) , freshly isolated T cells ( B , upper panel) and activated T cells [ (B) , lower panel]. Nuclei are stained with DAPI (blue). Images were taken with ×60 and ×2 zoom. (C) Colocalization of SIRPγ and CD47 on resting vs . activated T cells was investigated using ImageJ software and coloc2 plugin. Pearson value comprising between 0 (no colocalization) and 1 (colocalization) is presented for resting and activated T cells. n = 2 independent experiments.

Article Snippet: Recombinant protein , CD47 , AcroBiosystems, Newark, DE, USA , CD7-H82F6 , Human , .

Techniques: Confocal Microscopy, Imaging, Isolation, Staining, Software

Journal: Frontiers in Immunology

Article Title: SIRPγ-CD47 Interaction Positively Regulates the Activation of Human T Cells in Situation of Chronic Stimulation

doi: 10.3389/fimmu.2021.732530

Figure Lengend Snippet: SIRPγ polarizes at the immune synapse. (A) Time-lapse microscopy of the polarization of SIRPγ at the Raji-Jurkat cell contact. Jurkat cells were stained with the cell tracker (green) and an anti-SIRPγ (red), and Raji cells (not stained) were primed with SEE; white arrows indicate SIRPγ clusters at the Jurkat-Raji cell contact. (B) Similar culture condition as in (A) with several screenshots from a time-lapse video highlighting SIRPγ cluster at the Jurkat-Raji cell contact. (C) SEE-primed Raji cells were stained with an anti-CD47 mAb (green). After the interaction with Jurkat cells (blue), cells were fixed, permeabilized, and stained with phalloidin (red) which shows a polarization of actine at the Raji-Jurkat cell contact, indicating a synapse formation. Green staining also shows a polarization of the CD47 of Raji at the synapse. (D) Similar culture condition as in (A) but with T cells instead of Jurkat cells. T cells (blue) were stained with anti-SIRPγ mAb (white), and Raji cells were stained with anti-CD47 mAb (green). After the interaction, cells were fixed, permeabilized, and stained with phalloidin (red). Phalloidin staining shows the polarization of actine in the immune synapse as well as SIRPγ and CD47. (E) Description of the methodology is used to analyze the expression of SIRPγ regarding cell situation. (F) Histograms showing SIRPγ intensity on Jurkat cells interacting or not (control (CTL)) with SEE-primed Raji cells in the presence or not of KWAR23. n = 5 to 10 cells analyzed per group from three independent experiments. Statistical analysis was performed by a Mann-Whitney test; ** p ≤ 0.01; **** p ≤ 0.0001, ns, not significant.

Article Snippet: Recombinant protein , CD47 , AcroBiosystems, Newark, DE, USA , CD7-H82F6 , Human , .

Techniques: Time-lapse Microscopy, Staining, Expressing, Control, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: SIRPγ-CD47 Interaction Positively Regulates the Activation of Human T Cells in Situation of Chronic Stimulation

doi: 10.3389/fimmu.2021.732530

Figure Lengend Snippet: Inhibition of SIRPγ/CD47 interaction reduces IFN-γ secretion by chronically activated T cells. (A – C) T cells were stimulated once with coated OKT3 (0.5–1 µg/ml) with or without coated CD47-Fc (10 µg/ml) in the presence or not of KWAR23. (A) T-cell proliferation after 5 days of stimulation was analyzed by CPD dilution assay ( n = 3 independent experiments with a total of four different HV). (B) T-cell fitness was evaluated with Alamar Blue at Day 5; n = 4. (C) IFN-γ dosage in the supernatant of T cells after 2 days of stimulation in depicted stimulation conditions ( n = 12 within seven independent experiments). (A – C) Statistical analyses were performed with a one-way ANOVA test followed by Kruskal-Wallis; * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001, ns, not significant. (D, E) T cell subjected to three rounds of stimulation with coated OKT3 and anti-CD28 were further stimulated by a coated OKT3 stimulation with or without coated CD47-Fc (10 µg/ml). (D) T-cell fitness was evaluated with Alamar Blue at Day 5. (E) Supernatant were collected 48 h after the last stimulation and IFN-γ secretion was evaluated by ELISA. (D, E) Statistical analyses were performed by a Wilcoxon test. ns, not significant. (F, G) T cells subjected to three rounds of stimulation as previously mentioned were further stimulated during 48 h by a coated OKT3 stimulation with or without coated CD47-Fc (10 µg/ml) with various concentrations of KWAR23, B6H12, or LSB2.2O. T-cell fitness was evaluated with Alamar Blue, n = 2 (mean ± SEM). (G) Supernatants collected at 48 h were analyzed by ELISA. Statistical analyses were performed by a Kruskal-Wallis test. * p ≤ 0.05 between KWAR23 and ISO and KWAR23 and LSB2.20 when Ab were used at 1.1 µg/ml; * p ≤ 0.05 between KWAR23 and ISO when Ab were used at 0.37 µg/ml. n = 3–5 (mean ± SEM).

Article Snippet: Recombinant protein , CD47 , AcroBiosystems, Newark, DE, USA , CD7-H82F6 , Human , .

Techniques: Inhibition, Dilution Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: SIRPγ-CD47 Interaction Positively Regulates the Activation of Human T Cells in Situation of Chronic Stimulation

doi: 10.3389/fimmu.2021.732530

Figure Lengend Snippet: Inhibition of SIRPα-γ/CD47 interactions in vivo impairs human chimerism and delays xeno-GvHD. Recipient mice, irradiated at 1.5 Gy on Day 0, were injected i.v. with 10 million human PBMC 8 h later. Mice were i.p. treated 2×/week with the anti-SIRPα-γ mAb (KWAR23) at 5 mg/kg or with PBS until they reached notable clinical signs of xeno-GvHD. (A) Mice survival is shown regarding treatment. Data were analyzed with a Log-rank (Mantel-Cox) test. (B) Kinetics of human chimerism. Data were analyzed with a one-way ANOVA test followed by Kruskal-Wallis. (C) Percentage of human T cells within the human CD45+ population analyzed in the blood at D14; PBS-treated mice are presented in black, and KWAR23-treated mice are presented in red. Data were analyzed with a Mann-Whitney test. (A – C) Results from four experiments are presented in which human PBMC from four different healthy volunteers were injected in four to five mice per group. (D) CD4/CD8 ratio (percentage of positive cells within hCD45 population) was analyzed at D14 regarding the treatment. Data were analyzed with a Mann-Whitney test. n = 3 experiments with PBMC from three different HV donors (shown in different shape). (E) Absolute number of human CD4 and CD8 in the blood of recipient mice at Day 14. n = 3 experiments with PBMC from three different HV. Data were analyzed with a one-way ANOVA test followed by Kruskal-Wallis. (F) PD1 + cells within the circulating human CD4 and CD8 T cells; data were analyzed with a one-way ANOVA test. (G) IFN-γ dosage sera collected at Day 14 are shown; data were analyzed with a Mann-Whitney test. (H) Representative dot plots of human CD45 + CD3 + in blood of humanized mice treated with PBS or KWAR23; SIRPγ expression on CD4 + and CD8 T cells (i.e., CD4 − ) is presented. (I) Percentage of SIRPγ + CD4 + and SIRPγ + CD8 + cells are shown and were analyzed with a one-way ANOVA test. (J) SIRPγ geometric mean on human CD4 and CD8 T cells. Data were analyzed with a one-way ANOVA test followed by Kruskal-Wallis. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001; ns, not significant.

Article Snippet: Recombinant protein , CD47 , AcroBiosystems, Newark, DE, USA , CD7-H82F6 , Human , .

Techniques: Inhibition, In Vivo, Irradiation, Injection, MANN-WHITNEY, Expressing

Journal: Frontiers in Immunology

Article Title: SIRPγ-CD47 Interaction Positively Regulates the Activation of Human T Cells in Situation of Chronic Stimulation

doi: 10.3389/fimmu.2021.732530

Figure Lengend Snippet: Inhibition of SIRPα-γ/CD47 interactions alters blood memory cells in humanized mice. At Day 14, blood cells were analyzed by FACS. (A) Percentage of human CD4 and (B) CD8 subpopulations are presented based on FACS analysis with the following patterns CD45RA − CD27 − effector memory cells (EM), CD45RA − CD27 + central memory cells (CM), CD45RA + CD27 + -naïve cells, and CD4 − CD45RA + CD27 − for TEMRA CD8 cells. (C) Absolute number of each subpopulation of CD4 + and (D) CD8 + T cells are presented. n = 3 experiments with PBMC from three different HV. Data were analyzed with a one-way ANOVA test followed by Kruskal-Wallis. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001; ns, not significant.

Article Snippet: Recombinant protein , CD47 , AcroBiosystems, Newark, DE, USA , CD7-H82F6 , Human , .

Techniques: Inhibition

Journal: Frontiers in Immunology

Article Title: SIRPγ-CD47 Interaction Positively Regulates the Activation of Human T Cells in Situation of Chronic Stimulation

doi: 10.3389/fimmu.2021.732530

Figure Lengend Snippet: Inhibition of SIRPα-γ/CD47 interactions alters memory cells in the spleen of humanized mice. At Day 14, splenocytes were analyzed by FACS. (A) Human chimerism is shown; data were analyzed with a Mann-Whitney test. (B) CD4/CD8 ratio is shown regarding the treatment; data were analyzed with a Mann-Whitney test. (C) Relative number of human CD4 and CD8 in the splenocytes of recipient mice at Day 14. Data were analyzed with a one-way ANOVA test followed by Kruskal-Wallis. (D) Percentage of human CD4 and (E) CD8 subpopulations is presented based on FACS analysis with the following patterns CD45RA − CD27 − effector memory cells (EM), CD45RA − CD27 + central memory cells (CM), CD45RA + CD27 + -naïve cells, and CD4 − CD45RA + CD27 − for TEMRA CD8 cells. (F) Relative number of each subpopulation of CD4 + and (G) CD8 + T cells are presented. n = 1 experiment with PBMC from one HV. Data were analyzed with a one-way ANOVA test followed by Kruskal-Wallis. * p ≤ 0.05; ** p ≤ 0.01; ns, not significant.

Article Snippet: Recombinant protein , CD47 , AcroBiosystems, Newark, DE, USA , CD7-H82F6 , Human , .

Techniques: Inhibition, MANN-WHITNEY