Journal: International Journal of Nanomedicine

Article Title: Cathepsin B-Activatable Bioactive Peptide Nanocarrier for High-Efficiency Immunotherapy of Asthma

doi: 10.2147/IJN.S455633

Figure Lengend Snippet: Effect of PEG-Liker (PEG-IALLIPF), Trp, or MPP-Trp on the production of NO and Pro-inflammatory cytokines. ( A ) In Control-PBMCs, Asthma-PBMCs, Asthma-PBMCs + PEG-Liker, Asthma-PBMCs + Trp and Asthma-PBMCs + MPP-Trp group, NO production were examined by NO assay kit. Pro-inflammatory cytokines ( B ) TNF-α, ( C ) IL-1β, and ( D ) IL-6 contents in all groups were examined by ELISA assay. Data were presented as mean ± SD of three independent experiments. * P <0.05, ** P <0.01, *** P <0.001 vs Asthma-PBMCs group.

Article Snippet: The total peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood samples of control group and asthma group by mouse peripheral blood mononuclear cell isolation kit (P6340; Solarbio, Beijing, China) according to the manufacturer's protocol.

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Nanomedicine

Article Title: Cathepsin B-Activatable Bioactive Peptide Nanocarrier for High-Efficiency Immunotherapy of Asthma

doi: 10.2147/IJN.S455633

Figure Lengend Snippet: MPP-Trp was closely correlated with the balanced Th1/Th2 level and Th1/Th2-type cytokine production. ( A ) In Control-PBMCs, Asthma-PBMCs, Asthma-PBMCs + PEG-Liker, Asthma-PBMCs + Trp and Asthma-PBMCs + MPP-Trp group, Th1 population (CD4+IFN-γ+) and Th2 population (CD4+IL-4+) were selected by flow cytometry assay. ( B ) Relative mRNA expressions of IFN-γ, IL-4, IL-13, and IL-5 in all groups were assessed by qRT-PCR. ( C ) The IFN-γ, IL-4, IL-13, and IL-5 contents in all groups were examined by ELISA assay. Data were presented as mean ± SD of three independent experiments. * P <0.05, ** P <0.01, *** P <0.001 vs Asthma-PBMCs group.

Article Snippet: The total peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood samples of control group and asthma group by mouse peripheral blood mononuclear cell isolation kit (P6340; Solarbio, Beijing, China) according to the manufacturer's protocol.

Techniques: Control, Flow Cytometry, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Nanomedicine

Article Title: Cathepsin B-Activatable Bioactive Peptide Nanocarrier for High-Efficiency Immunotherapy of Asthma

doi: 10.2147/IJN.S455633

Figure Lengend Snippet: MPP-Trp altered cytokine gene expression and production in a concentration-dependent way. In Control-PBMCs, Asthma-PBMCs, and Asthma-PBMCs + MPP-Trp (10, 50, 100 and 200 μg/mL) group, ( A ) Th1/Th2 cytokine gene expressions (IFN-γ, IL-4, IL-13, and IL-5) were determined by RT-qPCR. ( B ) The cytokines (IFN-γ, IL-4, IL-13, and IL-5) levels in all groups were detected by ELISA. Data were presented as mean ± SD of three independent experiments. ** P <0.01, *** P <0.001 vs Control-PBMCs group; # P <0.05, ## P <0.01, ### P <0.001 vs Asthma-PBMCs group.

Article Snippet: The total peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood samples of control group and asthma group by mouse peripheral blood mononuclear cell isolation kit (P6340; Solarbio, Beijing, China) according to the manufacturer's protocol.

Techniques: Gene Expression, Concentration Assay, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Nanomedicine

Article Title: Cathepsin B-Activatable Bioactive Peptide Nanocarrier for High-Efficiency Immunotherapy of Asthma

doi: 10.2147/IJN.S455633

Figure Lengend Snippet: MPP-Trp altered cytokine gene expression and production in a time-dependent way. ( A ) In Control-PBMCs, Asthma-PBMCs, and 100 μg/mL Asthma-PBMCs + MPP-Trp (6, 12, 24, and 48 h) group, IFN-γ, IL-4, IL-13, and IL-5 mRNA levels were determined by RT-qPCR. ( B ) The Th1/Th2-type cytokines (IFN-γ, IL-4, IL-13, and IL-5) productions in all group were examined by ELISA. Data were presented as mean ± SD of three independent experiments. *** P <0.001 vs Control-PBMCs group; # P <0.05; ## P <0.01; ### P <0.001 vs Asthma-PBMCs group.

Article Snippet: The total peripheral blood mononuclear cells (PBMCs) were isolated from the whole blood samples of control group and asthma group by mouse peripheral blood mononuclear cell isolation kit (P6340; Solarbio, Beijing, China) according to the manufacturer's protocol.

Techniques: Gene Expression, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Engineered oncolytic virus expressing B7H3-targeting BiTE enhances antitumor T-cell immune response

doi: 10.1136/jitc-2024-009901

Figure Lengend Snippet: B7H3 expression in cell lines and the binding of delivered B7H3 BiTE to T and target cell. ( A ) B7H3 expression on MGC-803, SH-SY5Y, U87, SGC-7901 and Raji cells were detected using flow cytometry. ( B ) Binding of delivered B7H3 BiTE to target cells. Several cell lines were incubated with supernatants of the OAd-B7H3-BiTE-infected HEK293 cells. B7H3 BiTE binding was detected via flow cytometry with anti–anti-CD3-scFv antibody. ( C ) CD3 expression on Jurkat cells was detected using flow cytometry. ( D ) Binding of delivered B7H3 BiTE to T cells. PBMCs were incubated with supernatants of the OAd-B7H3-BiTE-infected HEK293 cells. B7H3 BiTE binding was detected via flow cytometry with anti–anti-CD3-scFv antibody. ( E ) Competition between delivered B7H3 BiTE and OKT3. PBMCs were incubated with supernatants of OAd or OAd-B7H3-BiTE-infected HEK293 cells. Then, cells were stained with fluorescein-conjugated OKT3 and examined throughflow cytometry. ( F ) Cell-bridging assay. A mixture of MGC-803/U87 and PBMCs was incubated with the supernatants of the OAd or OAd-B7H3-BiTE-infected HEK293 cells. The percentage of T/target cell pairs was quantified using flow cytometry. Statistical analysis was performed by unpaired Student’s t-test (n=3). NS: not significant; **p<0.01 and ****p<0.0001. Data are presented as the mean±SD. BiTE, bispecific T-cell engager; OAd, oncolytic adenovirus; PBMC, peripheral blood mononuclear cell; scFv, single-chain variable fragment.

Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy adults using a PBMC Isolation Solution Kit (P8680; Solarbio).

Techniques: Expressing, Binding Assay, Flow Cytometry, Incubation, Infection, Staining, Cytometry

Journal: Journal for Immunotherapy of Cancer

Article Title: Engineered oncolytic virus expressing B7H3-targeting BiTE enhances antitumor T-cell immune response

doi: 10.1136/jitc-2024-009901

Figure Lengend Snippet: T-cell activation mediated by oncolytic viruses-delivered B7H3 BiTE. ( A – D ) PBMCs were cultured alone or co-cultured with the indicated cell lines in the absence (Mock) or presence of the supernatants of OAd or OAd-B7H3-BiTE-infected cells for 48 hours. ( A ) Representative brightfield images are shown. ( B ) Representative flow cytometry histograms showing FSC-A of CD4 + and CD8 + T cells. ( C ) Cumulative data of flow cytometry showing CD69 or CD107a expression in CD4 + and CD8 + T cells. ( D ) The levels of IFN-γ and IL-2 in supernatants at 48 hours measured by ELISA. ( E ) T-cell proliferation assay. CFSE-stained PBMCs were co-cultured with MGC-803 cells and incubated with or without the supernatants of OAd or OAd-B7H3-BiTE-infected cells. Proliferation was determined after incubation for 4 days. Statistical analysis was performed by unpaired Student’s t-test (n=3). NS: not significant; **p<0.01, ***p<0.001, and ****p<0.0001. Data are presented as the mean±SD. BiTE, bispecific T-cell engager; CFSE, 5-(and 6)-carboxyfluorescein diacetate succinimidyl ester; IFN, interferon; IL, interleukin; OAd, oncolytic adenovirus; PBMC, peripheral blood mononuclear cell.

Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy adults using a PBMC Isolation Solution Kit (P8680; Solarbio).

Techniques: Activation Assay, Cell Culture, Infection, Flow Cytometry, Expressing, Enzyme-linked Immunosorbent Assay, Proliferation Assay, Staining, Incubation

Journal: Journal for Immunotherapy of Cancer

Article Title: Engineered oncolytic virus expressing B7H3-targeting BiTE enhances antitumor T-cell immune response

doi: 10.1136/jitc-2024-009901

Figure Lengend Snippet: T-cell cytotoxicity mediated by OAd-B7H3-BiTE. ( A – C ) After labeling with CFSE, MGC-803 ( A ), U87 ( B ) or Raji ( C ) cells were infected with OAd or OAd-B7H3-BiTE; uninfected cells (Mock) were used as a control. After 24 hours, peripheral blood mononuclear cells were added and co-cultured for 48 hours. Representative flow cytometry plots (left) and cumulative data (right) are shown. Dead target cells are defined as CFSE + Zombie Dye + . The percentage of dead target cells is calculated as follows: CFSE + Zombie-Dye + /CFSE + . Statistical analysis was performed by unpaired Student’s t-test (n=3). **p<0.01, ***p<0.001, and ****p<0.0001. Data are presented as the mean±SD. BiTE, bispecific T-cell engager; CFSE, 5-(and 6)-carboxyfluorescein diacetate succinimidyl ester; OAd, oncolytic adenovirus.

Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy adults using a PBMC Isolation Solution Kit (P8680; Solarbio).

Techniques: Labeling, Infection, Control, Cell Culture, Flow Cytometry

Journal: Journal for Immunotherapy of Cancer

Article Title: Engineered oncolytic virus expressing B7H3-targeting BiTE enhances antitumor T-cell immune response

doi: 10.1136/jitc-2024-009901

Figure Lengend Snippet: OAd-B7H3-BiTE treatment elicits potent therapeutic effects in vivo. ( A ) Schematic diagram of treatment schedule for MGC-803 bearing mice ( B – G ). ( B ) Representative flow cytometry plots showing human CD45 + CD3 + T cells in tumor and spleen of untreated mice. Flow cytometry plots gated on live-cell population. ( C ) Summary data for average tumor growth (left) and body weight changes (right) for all treatment groups. ( D ) Tumor pictures and weights for all treatment groups. ( E – F ) Primary tumors were collected and analyzed with reverse transcription-quantitative PCR to examine the expression of E1a ( E ) and B7H3 BiTE ( F ). ( G ) Representative H&E staining of the major organs for all treatment groups. Statistical analysis was performed by unpaired two-way analysis of variance ( C ) or unpaired Student’s t-test ( D ) (n=5). ***p<0.001, and ****p<0.0001. Data are presented as the mean±SD. BiTE, bispecific T-cell engager; mRNA, messenger RNA; OAd, oncolytic adenovirus; PBS, phosphate-buffered saline; PBMC, peripheral blood mononuclear cell; s.c., subcutaneous injection.

Article Snippet: Peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy adults using a PBMC Isolation Solution Kit (P8680; Solarbio).

Techniques: In Vivo, Flow Cytometry, Reverse Transcription, Real-time Polymerase Chain Reaction, Expressing, Staining, Saline, Injection

Journal: mAbs

Article Title: Structural and functional characterization of a monoclonal antibody blocking TIGIT.

doi: 10.1080/19420862.2021.2013750

Figure Lengend Snippet: Figure 4. Antitumor activity of MG1131 in cell-based assays. (A) Increased cytotoxic activity of NK cells. Expanded NK cells were co-cultured with the indicated target cells (stained with calcein AM) with an E:T ratio of 10:1, and the levels of fluorescence from the supernatant were measured as a function of the concentration of MG1131 or 1D3. N = 3. Bars: mean ± standard error of the mean. (B) Decreased expression of immune checkpoint proteins. Following T cell activation, Treg cells were treated with MG1131 or 1D3 at 10 μg/mL. The expression of the indicated immune checkpoint proteins on Treg cells was detected by flow cytometry. Treg cells were CD3+CD4+CD25hiCD127loFoxP3+ cells. Treg cells from nine different donors were used. *, p < .05; **, p < .01 by paired t-test. In A-C, human IgG4 was used as a control. (C) Decreased activity of Treg cells. CFSE-labeled Tresp cells were activated with anti-CD3/CD28 microbeads, treated with MG1131 or 1D3 at 10 μg/mL, and cultured with or without Treg cells for 5 days. The suppressive activity of Treg cells was calculated based on the proliferation of Tresp cells (see Materials and Methods). Five pairs of the marks (○, ■) represent five different donors. p < .05 by paired t-test.(D) Improved cytotoxic activities of CD8+ T cells derived from multiple myeloma patients. Following T cell activation with anti-CD3/CD28 microbeads, PBMCs from multiple myeloma patients were incubated with MG1131 or human IgG4 (control) at 10 μg/mL for 4 days. Secretion of IFN-γ was measured by flow cytometry. Results from five different donors (MM1-MM5) are shown. Bars: mean ± SEM. *, p < .05, ***, p < .005 by unpaired t-test.Figure 4a Alt text: The effect of MG1131 and 1D3 on the cell killing activity of natural killer cells were measured for three different cell lines. Line graphs show the increase in the cytotoxicity activity of natural killer cells as a function of the concentration of the two antibodies.Figure 4b Alt text: The effects of MG1131 and 1D3 on the expression of immune checkpoint proteins are shown on the left and right panel, respectively. The two antibodies decrease the expression of these proteins. Figure 4c Alt text: The effects of MG1131 and 1D3 on the suppression of regulatory T cells are shown on the left and right panel, respectively. In these experiments (in Figure 4a-4c), MG1131 was better than or at least comparable to 1D3.Figure 4d Alt text: A bar graph showing the secretion of IFN-γ in PBMCs derived from multiple myeloma patients. MG1131 treatment enhanced the IFN-γ secretion in comparison with the control IgG4 antibody.

Article Snippet: Measurement of IFN-γ secretion from multiple myeloma patient-derived PBMCs PBMCs from multiple myeloma patients (Axol and iQ Bioscience) were thawed and seeded at 1 × 106 cells/well.

Techniques: Activity Assay, Cell Culture, Staining, Fluorescence, Concentration Assay, Expressing, Activation Assay, Flow Cytometry, Control, Labeling, Derivative Assay, Incubation, Comparison

Journal: Journal of Virology

Article Title: Rescue of naïve porcine circovirus type 3 and its pathogenesis in CD pigs

doi: 10.1128/jvi.00341-25

Figure Lengend Snippet: Infection characteristics of PCV3. Detection of viral viremia in sera ( A ) and viral load in tissues ( B ) using a real-time PCR method. ( C ) Histopathological lesions in the lungs, lymph nodes (LNs), liver, and intestine 35 d post-infection, with lesion sites indicated by red arrows (400×). ( D ) At 35 d post-infection, immunohistochemical analysis of the heart, liver, lungs, lymph nodes (LNs), spleen, and intestines was performed. Brown-specific punctate deposits were observed following DAB staining (400×). ( E ) At 14 and 28 days post-infection, PBMCs were separately stimulated with ConA and VLP-PCV3-Cap. After stimulation with ConA, there was no significant difference in cell proliferation between the infected group and the control group. However, following stimulation with VLP-PCV3-Cap, there was a significant difference between the two groups, and cell proliferation was inhibited. The data are presented as the means ± SDs. * P < 0.05; ** P < 0.01.

Article Snippet: PBMCs were isolated using a porcine PBMC isolation kit (Solarbio), which achieved a cell viability of approximately 90%.

Techniques: Infection, Real-time Polymerase Chain Reaction, Immunohistochemical staining, Staining, Control

Journal: Journal of Virology

Article Title: Rescue of naïve porcine circovirus type 3 and its pathogenesis in CD pigs

doi: 10.1128/jvi.00341-25

Figure Lengend Snippet: Single-cell profiling of cell populations in PBMCs. UMAP visualization of cell types in PBMCs from the control group ( A ) and the PCV3-infected group ( B ). ( C ) UMAP visualization comparing the heterogeneity of cell types between the control group and the PCV3-infected group. ( D ) The distribution of each cell type in the control group and the PCV3-infected group was represented as percentages.

Article Snippet: PBMCs were isolated using a porcine PBMC isolation kit (Solarbio), which achieved a cell viability of approximately 90%.

Techniques: Control, Infection

Journal: Journal of Virology

Article Title: Rescue of naïve porcine circovirus type 3 and its pathogenesis in CD pigs

doi: 10.1128/jvi.00341-25

Figure Lengend Snippet: Reclassification and pseudotime analysis of T-cell subsets in PBMCs. UMAP and pie charts showing the cell subpopulations and proportions after T-cell repopulation in control ( A ) and PCV3-infected ( B ) PBMCs. ( C ) The heterogeneity of cell subpopulations between the control and infected groups was demonstrated with UMAP plots after re-clustering, and the number of cells in the different subpopulations is labeled with bar graphs. ( D ) Fitted-time analysis of the distribution of cell subpopulations across differentiation states. ( E ) Number of cells between samples in the five differentiation states.

Article Snippet: PBMCs were isolated using a porcine PBMC isolation kit (Solarbio), which achieved a cell viability of approximately 90%.

Techniques: Control, Infection, Labeling