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: 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