Journal: bioRxiv

Article Title: Respiratory mucosal vaccination of peptide-poloxamine-DNA nanoparticles provides complete protection against lethal SARS-CoV-2 challenge

doi: 10.1101/2022.05.29.493866

Figure Lengend Snippet: Enzyme-linked immunospot (ELISpot) analyses of a ) IFN-γ and b ) IL-4 spot-forming cells in pulmonary lymphocyte after re-stimulation with peptide pools of 14-mer overlapping peptides spanning the SARS-CoV-2 receptor binding domain (RBD) region. c ) CD4 + T cells and d ) CD8 + T cells in the lung were assayed for IFN-γ + expression by flow cytometry after re-stimulation with the SARS-CoV-2 RBD peptide pool. e ) CD4 + T cells in the lung were analyzed for IL-4 + expression via flow cytometry in the same way as describe above. f ) Percentage of CD4 + effector memory T cells (TEM) co-expressing CD44 hi and CD62L lo and central memory T cells (TCM) co-expressing CD44 hi and CD62L hi in the lung of mice 35 days after priming. g ) Percentage of CD8 + T EM and T CM in lung of mice 35 days after priming. h ) Percentage of CD4 + T RM and i ) CD8 + T RM in lung of mice 35 days after initial vaccination. Data in a-i represent mean ± SEM (n=5 biologically independent samples). One-way ANOVA with Dunnett’s post-hoc test was used to determine significance (** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: For surface markers, the cells were incubated with anti-mouse CD4 (11-0041-82, Invitrogen), anti-mouse CD8a (45-0081-82, Invitrogen), For intracellular cytokine staining, cells were stimulated with the overlapping peptide pool spanning of 14-mer peptides overlapping by nine amino acids from the SARS-CoV-2 RBD proteins (see Supplementary Notes) for 6 h at 37 °C, 5% CO 2 .

Techniques: Enzyme-linked Immunospot, Binding Assay, Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: Respiratory mucosal vaccination of peptide-poloxamine-DNA nanoparticles provides complete protection against lethal SARS-CoV-2 challenge

doi: 10.1101/2022.05.29.493866

Figure Lengend Snippet: a ) After pulmonary inoculation, pSpike/PP-sNp is able to penetrate efficiently through the physical and biological barriers at the airway mucosal site. b ) With the help of targeting moiety (ICAM-1/CD54 ligand) in PP-sNp, pSpike/PP-sNp is internalized and captured by airway epithelial cells (AECs) or pulmonary dendritic cells (DCs). The cationic moiety within PP-sNp mediates endosome escape and nucleus localization of pSpike, followed by the transcription and translation process. The SARS-CoV-2 derived antigens expressed in AECs are subsequently presented to other immune cells such as pulmonary DCs. AECs simultaneously orchestrate the adaptive immune responses via the transient secretion of cytokines. Mature pulmonary DCs then migrate to the bronchial-associated lymphoid tissues (e.g., mediastinal lymph node) and present antigens to naïve T cells and B cells for humoral and cellular immune responses. c ) Activated B cells proliferate and differentiate into antibody-secreting plasma cells to generate secretory IgA (sIgA) and systemic IgG antibodies, the former of which efficiently neutralizes invading SARS-CoV-2 within the upper and lower respiratory tracts. d ) Meanwhile, a portion of T cells obtains a tissue resident memory phenotype (TRM), enabling them to reside in the airway and respond rapidly when encountering SARS-CoV-2 virus. Other activated CD4 + or CD8 + T cells, including CD44 hi CD62L lo T cells, CD4 + IFN-γ T cells and CD8 + IFN-γ T cells, take part in the process of eliminating SARS-CoV-2 infections as well. The robust and comprehensive immunity conferred by pulmonary vaccination of pSpike/PP-sNp probably controls SARS-CoV-2 replication and removes the viruses at the initial sites of infection, thus protects the vaccinated mice from lung lesions and death.

Article Snippet: For surface markers, the cells were incubated with anti-mouse CD4 (11-0041-82, Invitrogen), anti-mouse CD8a (45-0081-82, Invitrogen), For intracellular cytokine staining, cells were stimulated with the overlapping peptide pool spanning of 14-mer peptides overlapping by nine amino acids from the SARS-CoV-2 RBD proteins (see Supplementary Notes) for 6 h at 37 °C, 5% CO 2 .

Techniques: Derivative Assay, Infection

Journal: bioRxiv

Article Title: Glycan Profiling Identifies Chondroitin-4-sulfate as a Biomarker for Platinum Response and Therapeutic Target in Ovarian Cancer

doi: 10.1101/2025.10.06.675352

Figure Lengend Snippet: ( A ) Carboplatin and Triplatin chemical structures. ( B and C ) Cytotoxicity of carboplatin and Triplatin on human OVTOKO and JHOC5 cancer cell lines; 1h treatment. ( D ) Flow cytometry of 200 nM and 50 nM rVAR2-V5 binding to ES2 wt cells, ES2 wt cells + chABC treatment, or Xylt1/Xylt2 KO cells. ( E and F ) Cytotoxicity of carboplatin and Triplatin in ES2 wt and Xylt1/Xylt2 KO cell lines; 1h treatment. ( G and H ). Platinum cellular accumulation in ES2 wt and Xylt1/Xylt2 KO cells treated with 10 µM carboplatin or Triplatin for 1, 2, 4, and 8h. Platinum content was measured by inductively coupled plasma mass spectroscopy (ICP-MS) and normalized by number of cells. ( I and J ). Platinum-DNA adducts in ES2 wt and Xylt1/Xylt2 KO cells treated with 10 µM carboplatin or Triplatin 4, and 8h. ( K and M ) ES2-luc wt and Xylt1/Xylt2 KO tumors were implanted on the left and right flanks mice. Tumors were harvested after 24h treatment with 40 mg/kg i.p Carboplatin or 0.3 mg/kg i.p. Triplatin. Tumors were digested in nitric acid and platinum measured by ICP-MS.

Article Snippet: Bound peptide was detected with mouse anti-V5 antibody (R960-25, ThermoFisher; 1:700 in casein buffer, 45 min, room temperature), followed by Leica post-primary rabbit anti-mouse antibody (8 min) and goat anti-rabbit HRP conjugate (8 min), with PBS washes between each step.

Techniques: Flow Cytometry, Binding Assay, Clinical Proteomics, Mass Spectrometry

Journal: bioRxiv

Article Title: Glycan Profiling Identifies Chondroitin-4-sulfate as a Biomarker for Platinum Response and Therapeutic Target in Ovarian Cancer

doi: 10.1101/2025.10.06.675352

Figure Lengend Snippet: Representative images of rVAR2-V5 and H&E OC PDX staining and Qupath analysis. (C) Qupath segmentation of rVAR2-V5 (+), rVAR2-V5 (-), and necrotic tumor area in OC PDX models. (D) Sensitivity of OC PDX models to Triplatin and carboplatin. OC PDX models were treated i.p. with carboplatin (40 mg/kg) or Triplatin (0.3 mg/kg) on days 0, 4 and 8 (orange arrows). * * p<0.01, * * * * p<0.0001, 2-way ANOVA, Tukey

Article Snippet: Bound peptide was detected with mouse anti-V5 antibody (R960-25, ThermoFisher; 1:700 in casein buffer, 45 min, room temperature), followed by Leica post-primary rabbit anti-mouse antibody (8 min) and goat anti-rabbit HRP conjugate (8 min), with PBS washes between each step.

Techniques: Staining

Journal: bioRxiv

Article Title: Glycan Profiling Identifies Chondroitin-4-sulfate as a Biomarker for Platinum Response and Therapeutic Target in Ovarian Cancer

doi: 10.1101/2025.10.06.675352

Figure Lengend Snippet: (A) UVA CHTN OC TMA; Representative samples of patient OC subtypes (clinical history unknown) and (B) normal tissues stained with rVAR2-V5 protein. (C) UVA CHTN OC TMA; Percentage of sample area staining positively for rVAR2-V5. Values are representative of the mean of 4 cores per patient sample. (D) UVA CHTN OC TMA; Percentage of TMA samples above the cut-off score. (E) UPenn CCC TMA samples; Percentage of sample area staining positively for rVAR2-V5.

Article Snippet: Bound peptide was detected with mouse anti-V5 antibody (R960-25, ThermoFisher; 1:700 in casein buffer, 45 min, room temperature), followed by Leica post-primary rabbit anti-mouse antibody (8 min) and goat anti-rabbit HRP conjugate (8 min), with PBS washes between each step.

Techniques: Staining