Journal: Pharmaceutics

Article Title: LPX-TI641, a Tim3/4 Agonist, Induces Long-Term Immune Tolerance in Multiple Sclerosis Models

doi: 10.3390/pharmaceutics17111402

Figure Lengend Snippet: LPX-TI641 Ameliorates Disease in the MOG 35–55 induced EAE mouse model. ( A ) Clinical scores of mice immunized with MOG 35–55 and treated at disease onset with varying doses of LPX-TI641 ( n = 10/group). ( B ) Comparison of clinical EAE scores in mice treated with GA (glatiramer acetate), natalizumab, or LPX-TI641 ( n = 10/group). ( C ) Clinical scores of EAE mice initially treated with natalizumab and subsequently switched to LPX-TI641 ( n = 10/group). ( D ) Microscopic imaging (10×) of Luxol Fast Blue staining of brain tissue from EAE mice treated with: ( i ) LPX-TI641 (Gr 6); ( ii ) a single dose of natalizumab (Gr 4); or ( iii ) two doses of natalizumab followed by LPX-TI641 (Gr 5). The LFB images are provided as qualitative illustrations of regions of demyelination and preserved myelin. Arrows highlight those regions for clarity.

Article Snippet: These mice either continued DMF (2 mg/mouse PO QD for 14 days), switched to natalizumab (100 μg/mouse IV every three days for three doses; MedChem Express, cat# HY-108831), or switched to LPX-TI641 (3 μg/mouse SC QD for 14 days) ( ).

Techniques: Comparison, Imaging, Staining

Journal: Pharmaceutics

Article Title: LPX-TI641, a Tim3/4 Agonist, Induces Long-Term Immune Tolerance in Multiple Sclerosis Models

doi: 10.3390/pharmaceutics17111402

Figure Lengend Snippet: LPX-TI641 Suppresses Relapse in the PLP-Induced EAE Mouse Model. ( A ) Clinical scores of mice immunized with PLP and treated on day 15 post-immunization with varying doses of LPX-TI641 ( n = 10/group). ( B ) Comparison of EAE clinical scores in mice treated with natalizumab or LPX-TI641 ( n = 10/group). ( C ) Clinical scores of EAE mice initially treated with natalizumab and subsequently switched to LPX-TI641 ( n = 10/group). ( D ) Flow cytometry plots showing CD4 + Foxp3 + T-reg expression in brain ( top ) and spleen ( bottom ) at day 21 post-immunization across different treatment groups.

Article Snippet: These mice either continued DMF (2 mg/mouse PO QD for 14 days), switched to natalizumab (100 μg/mouse IV every three days for three doses; MedChem Express, cat# HY-108831), or switched to LPX-TI641 (3 μg/mouse SC QD for 14 days) ( ).

Techniques: Comparison, Flow Cytometry, Expressing

Journal: Pharmaceutics

Article Title: LPX-TI641, a Tim3/4 Agonist, Induces Long-Term Immune Tolerance in Multiple Sclerosis Models

doi: 10.3390/pharmaceutics17111402

Figure Lengend Snippet: LPX-TI641 in an Escalation Treatment Paradigm in the MOG-Induced EAE Mouse Model. ( A ). Kaplan–Meier plot showing the efficacy of LPX-TI641 in delaying or preventing disease progression compared to dimethyl fumarate (DMF). ( B ) EAE clinical scores in mice treated with DMF vs. LPX-TI641 ( n = 10/group). ( C ) Clinical scores for mice treated with DMF followed by escalation to either LPX-TI641 or natalizumab ( n = 10/group). ( D ) Survival curves comparing treatment groups following therapeutic escalation paradigm.

Article Snippet: These mice either continued DMF (2 mg/mouse PO QD for 14 days), switched to natalizumab (100 μg/mouse IV every three days for three doses; MedChem Express, cat# HY-108831), or switched to LPX-TI641 (3 μg/mouse SC QD for 14 days) ( ).

Techniques: Biomarker Discovery

Journal: Pharmaceutics

Article Title: LPX-TI641, a Tim3/4 Agonist, Induces Long-Term Immune Tolerance in Multiple Sclerosis Models

doi: 10.3390/pharmaceutics17111402

Figure Lengend Snippet: LPX-TI641 in an Induction/Maintenance Paradigm in the MOG-Induced EAE Mouse Model. ( A ) Kaplan–Meier plot showing the efficacy of LPX-TI641 in controlling disease compared to natalizumab. ( B ) EAE clinical scores in mice treated with natalizumab or LPX-TI641 ( n = 10/group). ( C ) Survival curves for mice randomized to receive LPX-TI641 or natalizumab during disease progression. ( D ) Survival curves during maintenance therapy with LPX-TI641 or natalizumab. ( E ) Clinical scores of EAE in mice initially treated with natalizumab and then transitioned to maintenance therapy with either dimethyl fumarate (DMF) or LPX-TI641.

Article Snippet: These mice either continued DMF (2 mg/mouse PO QD for 14 days), switched to natalizumab (100 μg/mouse IV every three days for three doses; MedChem Express, cat# HY-108831), or switched to LPX-TI641 (3 μg/mouse SC QD for 14 days) ( ).

Techniques: Biomarker Discovery

Journal: Molecular Medicine

Article Title: Bleomycin induces senescence and repression of DNA repair via downregulation of Rad51

doi: 10.1186/s10020-024-00821-y

Figure Lengend Snippet: Increased senescence markers and reduced Rad51 Expression in vivo. A Schematic picture of the animal experiment. B Representative HE (Upper panel), Masson staining (Middle panel), and SA-β-Gal staining (Lower panel) of harvested mouse lung tissues. C Representative immunohistochemical analysis of Rad51 (brown) in lung sections. The control group showed strong positive staining for Rad51 compared to the BLM and BLM + NTZ + Anti-Ly6G group (Arrows). D Quantitative analysis of fibrosis was performed using the Ashcroft score. E The semi-quantitative assessment of immunohistochemistry was determined by ImageJ software. F The intensity of Rad51 immunofluorescence was quantified by ImageJ software. G Representative images of Rad51 expression in AECs by co-immunofluorescence staining with both Rad51 (green) and PDPN (red). Significance markers: *p < 0.05; ***p < 0.001 compared to control; n = 6

Article Snippet: Natalizumab (NTZ) and Anti-Mouse Ly-6G/Ly-6C Antibody (Anti-Ly6G) were procured from MedChemExpress (MCE, Shanghai, China).

Techniques: Expressing, In Vivo, Staining, Immunohistochemical staining, Control, Immunohistochemistry, Software, Immunofluorescence

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: Plasmids used in this study.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Plasmid Preparation, Selection, Negative Control

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: Biophysical properties of the scFv purified in this study.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Purification

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: Yields in mg/L/OD 600 of scFv expressed with CyDisCo (black bars) or without (white bars). Average OD 600 of the cultures was 22 ± 3 for those containing CyDisCo and 27 ± 3 for cultures without CyDisCo. Error bars show standard deviation, n = 3–4. scFv frameworks which have all four cysteines mutated to alanine are indicated as CA.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Standard Deviation

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: Relative yield of scFv expressed without CyDisCo compared to the yield from expression with CyDisCo, n = 3–4.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: CD spectra of scFv that were found to be soluble in the absence of disulfide bonds and comparisons with the disulfide bonded counterpart. (A) scFv trastuzumab; (B) scFv natalizumab; (C) scFv trastuzumab and natalizumab and the trastuzumab framework—natalizumab CDR hybrid scFv all produced with CyDisCo; (D) trastuzumab framework—natalizumab CDR hybrid scFv. scFv frameworks which have all four cysteines mutated to alanine are indicated as CA.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Circular Dichroism, Produced

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: (A) western blot showing lane 1—binding of scFv with trastuzumab CDRs to the extracellular part of HER2 and lane 2—control sample showing binding of scFv with trastuzumab CDRs to the anti-His-tag secondary antibody; (a) trastuzumab + CyDisCo, (b) trastuzumab − CyDisCo, (c) trastuzumab CA − CyDisCo, (d) natalizumab framework-trastuzumab CDRs + CyDisCo, (e) Maa48 framework-trastuzumab CDRs + CyDisCo; (B) analysis of full length ITGA4 (purchased from 1. Abnova and 2. OriGene) on non-reducing SDS-PAGE; (C) dot blot of scFv containing natalizumab CDRs binding to full length ITGA4; (D) representative example of surface plasmon resonance (Biacore) analysis of binding of parental scFv IgA 1 to β-tubulin peptide epitope.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Western Blot, Binding Assay, Control, SDS Page, Dot Blot, SPR Assay

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: K D determination for scFv with trastuzumab CDRs binding to HER2 fragment V529-P625.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: K D determination for scFv with IgA 1 CDRs binding to β-tubulin peptide.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: (A) natalizumab framework—trastuzumab CDRs hybrid scFv. (B) natalizumab scFv. (C) trastuzumab scFv. For panels A-C all scFv were made in the presence of CyDisCo. The insert panel in each case shows the derivative of the change in fluorescence signal. (D) Correlation between melting temperature (Tm) and the yield of purified scFv expressed with (●) or without (○) disulfide bonds. Only those scFv showing a single thermal denaturation step are plotted. Coefficient of correlation between Tm and the yield of scFv with disulfide bonds R 2 = 0.12 and without disulfide bonds R 2 = 0.56.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Fluorescence, Purification

Journal: PLoS ONE

Article Title: Complementarity determining regions and frameworks contribute to the disulfide bond independent folding of intrinsically stable scFv

doi: 10.1371/journal.pone.0189964

Figure Lengend Snippet: Thermal stability of the scFv.

Article Snippet: ScFv with swapped CDRs along with trastuzumab and natalizumab scFv with all cysteines mutated to alanine were synthetized by GenScript and cloned Nde I / BamH I into an expression vector in frame with a C-terminal GSH 6 -tag.

Techniques: Expressing