Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 1 EVs-associated LGALS3BP levels correlate with metastatatic lesions and increase accordingly over time in a pseudometastatic model of human neuroblastoma. (A) Representative scheme of pseudometastatic model: human neuroblastoma SKNAS cells were tail-vein injected in NSG mice and blood was collected after 7 (n = 2), 14 (n = 3), 21 (n = 3), and 28 (n = 2) days. (B) EVs-associated LGALS3BP levels detected in ELISA (pg/mL) per µg of EVs isolated from mouse serum after 7 (n = 2), 14 (n = 3), 21 (n = 3), or 28 (n = 2) days. Tumor free mice (n = 2) were considered as controls. (C) Immunohis tochemistry staining with hematoxylin/eosin for metastatic lesions and LGALS3BP levels in metastatic lesions over time. Scale bar: 250 μm. Number of metastases detected in (D) liver, (E) lung, and (F) kidney of NSG mice after 7 (n = 2), 14 (n = 3), 21 (n = 3), and 28 (n = 2) days

Article Snippet: Blots were then probed with anti-human LGALS3BP (goat polyclonal; 1:1000, #AF2226; R&D Systems), anti-β-actin (mouse monoclonal; 1:40000, #A5441; Sigma-Aldrich), anti-Alix (mouse monoclonal; 1:500, #MA1-83977; Invitrogen), anti-Tsg101 (mouse monoclonal; 1:1000, #MA1-23296; Invitrogen).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Isolation, Staining

Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 2 Circulating and EVs-associated LGALS3BP could function as early detection biomarker of neuroblastoma lesions in a human neuroblastoma pseu dometastatic model. (A) EVs-associated LGALS3BP levels detected in ELISA (pg/mL) per µg of EVs isolated from mouse serum collected 14 days after NSG mice (n = 6) were tail-vein injected with human neuroblastoma SKNAS cells. Tumor free mice (n = 2) were considered as controls. (B) Representative NTA profile of EVs isolated from mouse serum after 14 days from tail-vein injection of SKNAS cells. (C) Circulating LGALS3BP levels detected in ELISA (pg/mL) in mouse serum (diluted 1:4) after 14 days (n = 6). Tumor free mice (n = 2) were considered as controls. Number of metastases detected in (D) liver, (E) lung, and (F) kidney of NSG at 14 days post-injection

Article Snippet: Blots were then probed with anti-human LGALS3BP (goat polyclonal; 1:1000, #AF2226; R&D Systems), anti-β-actin (mouse monoclonal; 1:40000, #A5441; Sigma-Aldrich), anti-Alix (mouse monoclonal; 1:500, #MA1-83977; Invitrogen), anti-Tsg101 (mouse monoclonal; 1:1000, #MA1-23296; Invitrogen).

Techniques: Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Isolation, Injection

Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 3 Human LGALS3BP is overexpressed and secreted by engineered murine neuroblastoma NXS2 hLGALS3BP cells. (A) Histograms showing transcrip tional levels of human (hLGALS3BP) and murine (mLGALS3BP) LGALS3BP in NXS2 CTRL and hLGALS3BP cells normalized to housekeeping gene (β-actin). Values refer to two independent biological replicates. (B) Western blotting images showing intracellular protein levels of hLGALS3BP in NXS2 CTRL and hLGALS3BP cells. Equal amounts of protein were loaded for each sample. β-actin was used as loading control. Images are representative of two indepen dent biological replicates. (C) Secreted levels (ng/mL) of hLGALS3BP and mLGALS3BP assessed by ELISA assay in NXS2 CTRL and hLGALS3BP cells. Values refer to two independent biological replicates. (D) Table showing secreted LGALS3BP expression level in a panel of human cancer cell lines. (E) Confocal imaging of NXS2 CTRL and hLGALS3BP cells showing LGALS3BP (green) and nuclei (Hoechst). Scale bar: 10 μm. (F) Immunoblot showing EVs-associated hLGALS3BP and positive/negative exosomal markers expression in whole cell lysates (WCL) or EVs isolated from NXS2 CTRL or hLGALS3BP. Equal amount of protein were loaded for each sample.

Article Snippet: Blots were then probed with anti-human LGALS3BP (goat polyclonal; 1:1000, #AF2226; R&D Systems), anti-β-actin (mouse monoclonal; 1:40000, #A5441; Sigma-Aldrich), anti-Alix (mouse monoclonal; 1:500, #MA1-83977; Invitrogen), anti-Tsg101 (mouse monoclonal; 1:1000, #MA1-23296; Invitrogen).

Techniques: Western Blot, Control, Enzyme-linked Immunosorbent Assay, Expressing, Imaging, Isolation

Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 5 Targeted and effective anti-tumor activity of anti-LGALS3BP ADC 1959-sss/DM4 in murine syngeneic neuroblastoma model. (A) Tumor growth expressed as tumor volume (mm3) from first treatment of NXS2 CTRL or hLGALS3BP allografts in syngeneic A/J mice treated or not with ADC 1959-sss/ DM4 (10 mg/kg; twice weekly; three total i.v. injections). Arrows indicate treatments administration. 4 or 5 mice per group were considered. (B) Kaplan- Meier curve showing probability of survival in percentage of mice bearing NXS2 CTRL or hLGALS3BP allografts treated or not with ADC 1959-sss/DM4 from first treatment. Log-rank (Mantel-Cox) test: ** p < 0.01. (C) TILs flow cytometric analysis of PBS (n = 4), 1959-sss (n = 2) and 1959-sss/DM4 (n = 4) treated tumors at 7 days after starting treatments, showing percentage of activated CD69 + within the total population of CD3+ T cells (left panel); percentage of activated CD8+ CD69+ within the total population of CD3+ T cells (middle panel). In the right panel is shown the percentage of CD69+ Granzyme B+ over cytotoxic CD8+ T cells. Differences were compared using unpaired T test, and considering significant p < 0.05, as indicated with an asterisk (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: Blots were then probed with anti-human LGALS3BP (goat polyclonal; 1:1000, #AF2226; R&D Systems), anti-β-actin (mouse monoclonal; 1:40000, #A5441; Sigma-Aldrich), anti-Alix (mouse monoclonal; 1:500, #MA1-83977; Invitrogen), anti-Tsg101 (mouse monoclonal; 1:1000, #MA1-23296; Invitrogen).

Techniques: Activity Assay

Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 6 Synergistic and durable anti-tumor activity of combination therapy based on anti-LGALS3BP ADC 1959-sss/DM4 and immune check-point inhibi tor anti-PD-1 in murine syngeneic neuroblastoma model. (A) Tumor growth expressed as tumor volume (mm3) from first treatment of NXS2 hLGALS3BP allografts in syngeneic A/J mice treated or not with anti-PD-1 (10 mg/kg; twice weekly; five total i.p. injections), naked 1959-sss (10 mg/kg; twice weekly; three total i.v. injections), ADC 1959-sss/DM4 (10 mg/kg; twice weekly; three total i.v. injections), or combination of 1959-sss and anti-PD-1, or combina tion of 1959-sss/DM4 and anti-PD-1. Blue and pink arrows indicate 1959-sss or 1959-sss/DM4 and anti-PD-1 treatments administration, respectively. 5 or 6 mice were considered per experimental group. (B) Kaplan-Meier curve showing probability of survival of mice bearing NXS2 hLGALS3BP allografts treated or not with anti-PD-1, 1959-sss, ADC 1959-sss/DM4, combination of 1959-sss and anti-PD-1, or combination of 1959-sss/DM4 and anti-PD-1 from first treatment. Log-rank (Mantel-Cox) test: *** p < 0.001

Article Snippet: Blots were then probed with anti-human LGALS3BP (goat polyclonal; 1:1000, #AF2226; R&D Systems), anti-β-actin (mouse monoclonal; 1:40000, #A5441; Sigma-Aldrich), anti-Alix (mouse monoclonal; 1:500, #MA1-83977; Invitrogen), anti-Tsg101 (mouse monoclonal; 1:1000, #MA1-23296; Invitrogen).

Techniques: Activity Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Convergent and Divergent Pathways Decoding Hierarchical Additive Mechanisms in Treating Cerebral Ischemia–Reperfusion Injury

doi: 10.1111/cns.12205

Figure Lengend Snippet: Additive mechanism of pathway convergence and divergence in Huntington's disease signaling pathway. On the basis of Ingenuity Pathway Analysis (IPA) knowledge base, the overlapping components between T Cell Receptor Signaling of baicalin (BA) group and Huntington's disease signaling of BJ group were Ca2+, PIP3, c‐Jun, JNK1, PIP2, IP3, DAG, PI3K, GRB2, SOS, hRas, and ERK/1/2; those between ERK/MAPK signaling of jasminoidin (JA) group and Huntington's disease signaling of BJ group were CREB, SHC, PIP2, IP3, DAG, PI3K, GRB2, SOS, hRas, and ERK/1/2; and the overlapping ones among the three groups were PIP2, IP3, DAG, PI3K, GRB2, SOS, hRas, and ERK/1/2. Differentially expressed genes in the three groups are marked with pink, blue, green, and yellow arrows, up‐ and down‐regulated genes were labeled with the up and down arrow, respectively. The overlapping components between the different groups are marked with different color stars.

Article Snippet: Blots were stained with the mice anticyclic adenosine monophosphate cAMP‐response‐element‐binding protein (CREB) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), which was developed by enhanced chemiluminescence (Amersham).

Techniques: Labeling

Journal: CNS Neuroscience & Therapeutics

Article Title: Convergent and Divergent Pathways Decoding Hierarchical Additive Mechanisms in Treating Cerebral Ischemia–Reperfusion Injury

doi: 10.1111/cns.12205

Figure Lengend Snippet: Validation of Rgs6, SH2‐B, cAMP‐response‐element‐binding protein (CREB), p‐ERK1/2 expression levels using both real‐time polymerase chain reaction (RT‐PCR) and Western blot analysis. (A) No significant discrepancies were found between the findings of microarray and real‐time RT‐PCR analyses. (B) Western blot analysis was employed to identify CREB and p‐ERK1/2 between different groups. *P < 0.05 versus vehicle.

Article Snippet: Blots were stained with the mice anticyclic adenosine monophosphate cAMP‐response‐element‐binding protein (CREB) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), which was developed by enhanced chemiluminescence (Amersham).

Techniques: Biomarker Discovery, Binding Assay, Expressing, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Western Blot, Microarray, Quantitative RT-PCR