Journal: Molecular Therapy Advances

Article Title: Efficacy and safety of SENS-501, a dual-AAV otoferlin gene therapy, for DFNB9 congenital deafness

doi: 10.1016/j.omta.2026.201762

Figure Lengend Snippet: Intracochlear injection of SENS-501 in NHPs results in a mild humoral and an undetectable cellular response to the capsid (A) AAV8 anti-drug antibody (ADA) titers measured in serum at pre-dose and at 16, 29, and 92 days post-vector administration in NHPs. (B) Anti-AAV8 neutralizing antibodies (NAb) titers measured in serum at pre-dose and at 16, 29, and 92 days post-vector administration in NHPs. (C and D) IFN-γ spot forming units (SFUs) measured by ELISpot assay at 29 (C; left) and 92 (D; right) days post-injection. Peripheral blood mononuclear cells (PBMCs) from the indicated groups were stimulated with three different AAV8 peptide pools and a positive control (PMA/ionomycin). The dotted line represents the assay-specific positivity threshold. Each dot represents one animal. Bars represent the mean ± SEM.

Article Snippet: After the incubation, detection was performed with a monoclonal anti-monkey IFN-γ antibody (Monkey IFN-γ ELISpot Pro Kit, Mabtech) coupled with alkaline phosphatase and incubated with BCIP/NBT (5-bromo-4-chloro-3-indolyl-1-phosphate / nitroblue tetrazolium) substrate to detect secreted IFN-γ.

Techniques: Injection, Plasmid Preparation, Enzyme-linked Immunospot, Positive Control

Journal: Poultry Science

Article Title: A novel self-amplified RNA vaccine co-expressing NA and HA1 delivered by Salmonella confers potent protection against H9N2 influenza in chickens

doi: 10.1016/j.psj.2026.107072

Figure Lengend Snippet: Intracellular cytokine production. Chicken splenic lymphocytes were isolated for analysis. Cell proliferation was assessed using CCK-8 analysis with ConA (A), mixed HA1 peptides (B), and NA protein (C). Additionally, the production of IFN-γ by splenic T lymphocytes was measured via an ELISpot assay, utilizing NA and HA1 proteins as stimulators for 36 h (D).

Article Snippet: IFN-γ production was assessed using a commercial Chicken IFN-γ ELISpot kit (Mabtech, Sweden).

Techniques: Isolation, CCK-8 Assay, Enzyme-linked Immunospot

Journal: Poultry Science

Article Title: A novel self-amplified RNA vaccine co-expressing NA and HA1 delivered by Salmonella confers potent protection against H9N2 influenza in chickens

doi: 10.1016/j.psj.2026.107072

Figure Lengend Snippet: Intracellular cytokine production. The intracellular mRNA expression levels of IL-4 (B, D) and IFN-γ (A, C)—as well as the relative concentrations of these cytokines in cell culture supernatants stimulated by the NA peptide (E, F) or HA1 protein (G, H) for 48 h—were determined using qRT-PCR and ELISA, respectively. Data are expressed as the mean ± SEM and analyzed using one-way ANOVA (* P < 0.05, ** P < 0.01, and *** P < 0.001; n = 4).

Article Snippet: IFN-γ production was assessed using a commercial Chicken IFN-γ ELISpot kit (Mabtech, Sweden).

Techniques: Expressing, Cell Culture, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vivo DC maturation and T cell activation in C57BL/6J mice induced by mOVA/H 18 NPs through intravenous injection. C57BL/6J mice were vaccinated with different formulations on Day 0 and Day 5. On Day 10, the spleens of mice were collected and analyzed by flow cytometry. Quantification analysis of (A) CD80 + CD86 + DCs and (B) CD40 + DCs in the spleen. Quantification analysis of (C) CD3 + CD4 + T cells and (D) CD3 + CD8 + T cells in the spleen. (E) Quantification analysis and (F) representative flow cytometry contour plots of OVA-specific CD8 + T cells among all cell populations in the spleen. (G) Quantification analysis and (H) representative flow cytometry contour plots of IFN-γ + CD8 + T cells among all cell populations in the spleen. (I) Quantification results and (J) representative images of IFN- γ -secreting immune cells in the spleen of mice analyzed by enzyme-linked immunospot (ELISpot) assay. Data were shown as mean ± SD (n = 3).

Article Snippet: Quantification analysis of (C) CD3 + CD4 + T cells and (D) CD3 + CD8 + T cells in the spleen. (E) Quantification analysis and (F) representative flow cytometry contour plots of OVA-specific CD8 + T cells among all cell populations in the spleen. (G) Quantification analysis and (H) representative flow cytometry contour plots of IFN-γ + CD8 + T cells among all cell populations in the spleen. (I) Quantification results and (J) representative images of IFN- γ -secreting immune cells in the spleen of mice analyzed by enzyme-linked immunospot (ELISpot) assay.

Techniques: In Vivo, Activation Assay, Injection, Flow Cytometry, Enzyme-linked Immunospot

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vitro effects of mOVA/H 18 NPs on activation of BMDCs. BMDCs were incubated with different formulations for 24 h and analyzed by flow cytometry. Quantification analysis for CD80 + CD86 + cells (A) and CD40 + cells (C) in BMDCs. Representative flow cytometry contour plots (B) for CD80 + CD86 + cells and histograms (D) for CD40 + cells in BMDCs. Concentrations of IL-4 (E), TNF-α (F), IFN-γ (G) and IL-12 (H) in BMDCs medium detected using ELISA. Data were shown as mean ± SD (n = 3).

Article Snippet: Mouse IFN-γ precoated ELISPOT kit was purchased from DAKEWE (Beijing, China).

Techniques: In Vitro, Activation Assay, Incubation, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: In vivo DC maturation and T cell activation in C57BL/6J mice induced by mOVA/H 18 NPs through intravenous injection. C57BL/6J mice were vaccinated with different formulations on Day 0 and Day 5. On Day 10, the spleens of mice were collected and analyzed by flow cytometry. Quantification analysis of (A) CD80 + CD86 + DCs and (B) CD40 + DCs in the spleen. Quantification analysis of (C) CD3 + CD4 + T cells and (D) CD3 + CD8 + T cells in the spleen. (E) Quantification analysis and (F) representative flow cytometry contour plots of OVA-specific CD8 + T cells among all cell populations in the spleen. (G) Quantification analysis and (H) representative flow cytometry contour plots of IFN-γ + CD8 + T cells among all cell populations in the spleen. (I) Quantification results and (J) representative images of IFN- γ -secreting immune cells in the spleen of mice analyzed by enzyme-linked immunospot (ELISpot) assay. Data were shown as mean ± SD (n = 3).

Article Snippet: Mouse IFN-γ precoated ELISPOT kit was purchased from DAKEWE (Beijing, China).

Techniques: In Vivo, Activation Assay, Injection, Flow Cytometry, Enzyme-linked Immunospot

Journal: Bioactive Materials

Article Title: Splenic dendritic cell-targeting mRNA transfection of H-type ionizable lipid-based LNPs for enhancing tumor immunotherapy

doi: 10.1016/j.bioactmat.2026.02.018

Figure Lengend Snippet: Anti-tumor effects of mTrp2/H 18 NPs as therapeutic vaccines in vivo and inhibitory effects of mOVA/H 18 NPs on lung metastasis of B16-OVA. (A) Schematic illustration of experiment design. B16F10 cells were inoculated subcutaneously on C57BL/6J mice on Day 0. The B16F10 bearing mice were vaccinated on Day 5 and Day 10 through intravenous injection. On Day 16, the mice were sacrificed for further flow cytometry analysis. mTrp2/MC3-LNP was administered at a mTrp2 dose of 0.75 mg kg −1. As for mTrp2/H 18 NPs, the doses were set at 0.25 mg kg −1 for mTrp2/H 18 NPs (L), 0.5 mg kg −1 for mTrp2/H 18 NPs (M), and 0.75 mg kg −1 for mTrp2/H 18 NPs (H). (B) Tumor growth curves of B16F10-bearing mice after treatment with different formulations (n = 6). (C) Survival curves of B16F10-bearing mice treated with different formulations (n = 6). The survival rates of the two groups were analyzed using a log-rank test. Quantification analysis of IFN-γ + cells among CD3 + CD8 + T cells (D) in the spleen, (E) in tumor tissues, and (F) in the blood (n = 3).

Article Snippet: Mouse IFN-γ precoated ELISPOT kit was purchased from DAKEWE (Beijing, China).

Techniques: Vaccines, In Vivo, Injection, Flow Cytometry

Journal: iScience

Article Title: Distinct CD8 + T cell types associated with COVID-19 severity in unvaccinated HLA-A2 + patients

doi: 10.1016/j.isci.2026.115880

Figure Lengend Snippet: Information on 11 selected HLA-A2-binding 9-mer peptides derived from the SARS-CoV-2 genome (A) Workflow of HLA-A2-binding peptide prediction with NETMHCpan4.1 and IFN-γ ELISpot assay of PBMCs from 24 patients with COVID-19 using 31 predicted peptides ( and ). (B) Homology of 11 selected peptides among SARS-CoV-2 variants and other strains (as indicated). “YES” indicates that more than 90% of epitope sequences matched within 200 selected peptide sequences from the database . Otherwise, the % of matched sequences is shown. “NO” indicates no matched sequences. (C) Information on the SARS-CoV-2 genome and mapping of 11 selected peptides on its genome.

Article Snippet: anti-human-IFN-γ antibody , MABTECH , Cat# 3420-3-250; RRID: AB_907283.

Techniques: Binding Assay, Derivative Assay, Enzyme-linked Immunospot

Journal: iScience

Article Title: Distinct CD8 + T cell types associated with COVID-19 severity in unvaccinated HLA-A2 + patients

doi: 10.1016/j.isci.2026.115880

Figure Lengend Snippet: IFN-γ ELISpot of CD8 + T cells in PBMCs from HLA-A2 + patients with COVID-19 using selected SARS-CoV-2 peptides (A) Cross-sectional ELISpot assay of HLA-A2-restricted CD8 + T cell responses against 11 selected peptides in PBMCs from 26 patients with mild, 8 with moderate, and 8 with severe COVID-19, respectively. (B) Log10-transformed mean spot-forming cell (SFC) counts (log10[Mean SFC +1]) per patient across 11 peptides in patients with mild, moderate, and severe COVID-19. Each point represents the mean response for one patient across 11 peptides ( n = 42 patients). A +1 pseudocount is added before the log10 transformation to accommodate zero values. Statistical comparisons were performed using a negative binomial mixed-effects model with Holm-Bonferroni correction for multiple comparisons. NS: not significant. (C) Longitudinal ELISpot assay of HLA-A2-restricted CD8 + T cell responses against selected peptides in PBMCs from patients with mild and moderate COVID-19 (as indicated).

Article Snippet: anti-human-IFN-γ antibody , MABTECH , Cat# 3420-3-250; RRID: AB_907283.

Techniques: Enzyme-linked Immunospot, Transformation Assay