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Mabtech Inc chicken ifn γ elispot kit
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 <t>of</t> <t>IFN-γ</t> by splenic T lymphocytes was measured via an <t>ELISpot</t> assay, utilizing NA and HA1 proteins as stimulators for 36 h (D).

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 <t>of</t> <t>IFN-γ</t> by splenic T lymphocytes was measured via an <t>ELISpot</t> assay, utilizing NA and HA1 proteins as stimulators for 36 h (D).

Chicken Ifn γ Elispot Kit, supplied by Mabtech Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 86 stars, based on 1 article reviews

chicken ifn γ elispot kit - by Bioz Stars, 2026-05

86/100 stars

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1) Product Images from "A novel self-amplified RNA vaccine co-expressing NA and HA1 delivered by Salmonella confers potent protection against H9N2 influenza in chickens"

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

Journal: Poultry Science

doi: 10.1016/j.psj.2026.107072

Figure Legend 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).

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

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).

Figure Legend 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).

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

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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

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).

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

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).

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

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

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).

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

Vectorized IFNβ drives durable signaling and complete tumor regression in human glioblastoma models in vivo (A) Sustained hIFNβ secretion in human GBM6 cells treated with AAV9-hIFNβ (red, MOI = 4E5 vg/cell) or recombinant hIFNβ cytokine (r-hIFNβ, purple, 47 IU/mL, equivalent to 114 pg/mL), measured by ELISA at indicated time points. 50% media washouts every 5 h for the first 20 h in the r-hIFNβ condition mimic in vivo cytokine clearance (half-life = 4–5 h). Full media exchanges were performed at 24, 48, 72, and 96 h post-treatment. (B) Number of differentially expressed genes (DEGs, p -Adj<0.01) in GBM6 cells 24–96 h post-treatment with AAV9-hIFNβ or r-hIFNβ vs. media controls. (C) Enrichment scores for type I IFN and TNFα response pathways across treatments and time points. (D) Heatmap of the top 10 IFN and TNFα response genes (Log2FC vs. media controls) in GBM6 cells treated as in (A). (E) Schematic of orthotopic PDX (SF11411) and cell line-derived xenograft ([CDX], GBM6-FLuc) studies in athymic nu/nu mice treated intratumorally with saline, AAV9-GFP, or AAV9-hIFNβ via CED. (F) Kaplan-Meier survival curves for PDX mice treated as in (E). Saline = black, AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ (2E11 vg/brain) = red. Vertical dashed line = day of treatment (day 9). p < 0.04 by log-rank (Mantel-Cox) test. n = 30 (10 per treatment arm). (G) Longitudinal BLI of GBM6-FLuc tumor growth in CDX mice treated as in (E). Saline = black, AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ (2E11 vg/brain) = red. Thin lines = individual mice, thick lines = geometric mean. Vertical dashed line = day of treatment (day 9). ∗ p < 0.04 by Kruskal-Wallis test with Dunn’s multiple comparisons correction on day 22. n = 30 (10 per treatment arm). (G′) Representative BLI images from each treatment group 11 days post-treatment. (H) Kaplan-Meier survival curves for CDX mice. p < 0.001 by log-rank (Mantel-Cox) test. (I) Distribution of treatment responses in CDX by BLI flux (photons/second) at day 27. Tumor free = BLI flux <2.5 × 10 5 p/s, tumor reduction = ≥30% decrease from assignment on day 9, no change = between 30% decrease and 20% increase from assignment on day 9, tumor growth = ≥20% increase from assignment on day 9, death = mice that died before day 27. (J) Dose-response analysis of AAV9-hIFNβ efficacy in CDX mice. AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ hi (2E11 vg/brain) = solid red, and AAV9-hIFNβ lo (1E11 vg/brain) = dashed red. Thin lines = individual mice, thick lines = geometric mean. Vertical dashed line = day of treatment (day 9). ∗∗ p < 0.02 by Kruskal-Wallis test with Dunn’s multiple comparisons correction on day 20. n = 45 (15 per treatment arm). For data interpretation, tumor burden threshold = 2.5 × 10 5 . (J′) Representative BLI images of tumors 11 days post-treatment. (K) Kaplan-Meier survival curves from (J). p < 0.002 (AAV9-hIFNβ hi), p < 0.005 (AAV9-hIFNβ lo) by log-rank (Mantel-Cox) test compared to AAV9-GFP. (I) Distribution of treatment responses in CDX mice at day 27 by BLI flux as in (I).

Journal: Molecular Therapy Oncology

Article Title: AAV immuno-gene therapy platform delivering vectorized cytokines defines a new modality for high-grade glioma treatment

doi: 10.1016/j.omton.2026.201183

Figure Lengend Snippet: Vectorized IFNβ drives durable signaling and complete tumor regression in human glioblastoma models in vivo (A) Sustained hIFNβ secretion in human GBM6 cells treated with AAV9-hIFNβ (red, MOI = 4E5 vg/cell) or recombinant hIFNβ cytokine (r-hIFNβ, purple, 47 IU/mL, equivalent to 114 pg/mL), measured by ELISA at indicated time points. 50% media washouts every 5 h for the first 20 h in the r-hIFNβ condition mimic in vivo cytokine clearance (half-life = 4–5 h). Full media exchanges were performed at 24, 48, 72, and 96 h post-treatment. (B) Number of differentially expressed genes (DEGs, p -Adj<0.01) in GBM6 cells 24–96 h post-treatment with AAV9-hIFNβ or r-hIFNβ vs. media controls. (C) Enrichment scores for type I IFN and TNFα response pathways across treatments and time points. (D) Heatmap of the top 10 IFN and TNFα response genes (Log2FC vs. media controls) in GBM6 cells treated as in (A). (E) Schematic of orthotopic PDX (SF11411) and cell line-derived xenograft ([CDX], GBM6-FLuc) studies in athymic nu/nu mice treated intratumorally with saline, AAV9-GFP, or AAV9-hIFNβ via CED. (F) Kaplan-Meier survival curves for PDX mice treated as in (E). Saline = black, AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ (2E11 vg/brain) = red. Vertical dashed line = day of treatment (day 9). p < 0.04 by log-rank (Mantel-Cox) test. n = 30 (10 per treatment arm). (G) Longitudinal BLI of GBM6-FLuc tumor growth in CDX mice treated as in (E). Saline = black, AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ (2E11 vg/brain) = red. Thin lines = individual mice, thick lines = geometric mean. Vertical dashed line = day of treatment (day 9). ∗ p < 0.04 by Kruskal-Wallis test with Dunn’s multiple comparisons correction on day 22. n = 30 (10 per treatment arm). (G′) Representative BLI images from each treatment group 11 days post-treatment. (H) Kaplan-Meier survival curves for CDX mice. p < 0.001 by log-rank (Mantel-Cox) test. (I) Distribution of treatment responses in CDX by BLI flux (photons/second) at day 27. Tumor free = BLI flux <2.5 × 10 5 p/s, tumor reduction = ≥30% decrease from assignment on day 9, no change = between 30% decrease and 20% increase from assignment on day 9, tumor growth = ≥20% increase from assignment on day 9, death = mice that died before day 27. (J) Dose-response analysis of AAV9-hIFNβ efficacy in CDX mice. AAV9-GFP (2E11 vg/brain) = blue, AAV9-hIFNβ hi (2E11 vg/brain) = solid red, and AAV9-hIFNβ lo (1E11 vg/brain) = dashed red. Thin lines = individual mice, thick lines = geometric mean. Vertical dashed line = day of treatment (day 9). ∗∗ p < 0.02 by Kruskal-Wallis test with Dunn’s multiple comparisons correction on day 20. n = 45 (15 per treatment arm). For data interpretation, tumor burden threshold = 2.5 × 10 5 . (J′) Representative BLI images of tumors 11 days post-treatment. (K) Kaplan-Meier survival curves from (J). p < 0.002 (AAV9-hIFNβ hi), p < 0.005 (AAV9-hIFNβ lo) by log-rank (Mantel-Cox) test compared to AAV9-GFP. (I) Distribution of treatment responses in CDX mice at day 27 by BLI flux as in (I).

Article Snippet: 24, 48, 72, and 96 h after treatment, cell supernatants were collected and IFN variant levels were measured using IFN ELISA kits following the manufacturer’s instructions (human IFNα [PBL Cat# 41135-1], human IFNβ [PBL Cat#41410], and human IFNγ [R&D Systems Cat#: DIF50C]).

Techniques: In Vivo, Recombinant, Enzyme-linked Immunosorbent Assay, Derivative Assay, Saline

Spatial transcriptomics reveals rapid, localized transcriptional remodeling of the tumor microenvironment following vectorized hIFNβ treatment (A) Coronal brain sections from representative human GBM6-FLuc CDX mice collected pre-treatment (0 h, n = 1) or 48 h ( n = 1) after intratumoral AAV9-hIFNβ infusion (2E11 vg/brain), stained with H&E (left) and subjected to Visium Spatial Gene Expression profiling (right). Annotated clusters were assigned based on anatomical localization and marker gene expression. Dashed lines denote tumor borders. Scale bars, 1 mm. (B) Top 10 marker genes for each spatially resolved cluster identified across 0 h and 48 h datasets. Values are shown as log-normalized expression centered at 0 (Seurat “scale.data”). (C) Spatial expression of canonical human GBM tumor markers ( CD44 , VIM , TOP2A , and NOTCH1 ) delineating tumor and peri-tumor regions before (0 h) (top) and after (bottom) (48 h) AAV9-hIFNβ treatment. (D) Expression maps of the human IFNβ payload and hallmark IFN-response genes ( CXCL10 , IFIT1 , and IFIT2 ), demonstrating tumor-restricted transgene expression and induction of an IFN-specific transcriptional program within 48 h. (E) Spatial expression of host mouse immune-response genes ( Gfap , Ifitm3 , and Irf7 ) showing localized activation of astroglial and innate immune pathways proximal to the tumor. (F) Integrated datasets (0 and 48 h) visualized using canonical correlation analysis (CCA), showing distinct clustering of tumor and peri-tumor regions (left) and enrichment of IFN-response gene module expression (right). (G) Volcano plot depicting differential gene expression between 0- and 48-h tumor clusters. Red, IFN-response genes; gray, other significantly upregulated genes ( p -Adj <0.01); blue, non-significant. (H) Top enriched Gene Ontology (GO) terms among upregulated genes in 48-h tumor cells, highlighting interferon and inflammatory response pathways (∗∗ p -Adj <0.01; ∗ p -Adj <0.05).

Journal: Molecular Therapy Oncology

Article Title: AAV immuno-gene therapy platform delivering vectorized cytokines defines a new modality for high-grade glioma treatment

doi: 10.1016/j.omton.2026.201183

Figure Lengend Snippet: Spatial transcriptomics reveals rapid, localized transcriptional remodeling of the tumor microenvironment following vectorized hIFNβ treatment (A) Coronal brain sections from representative human GBM6-FLuc CDX mice collected pre-treatment (0 h, n = 1) or 48 h ( n = 1) after intratumoral AAV9-hIFNβ infusion (2E11 vg/brain), stained with H&E (left) and subjected to Visium Spatial Gene Expression profiling (right). Annotated clusters were assigned based on anatomical localization and marker gene expression. Dashed lines denote tumor borders. Scale bars, 1 mm. (B) Top 10 marker genes for each spatially resolved cluster identified across 0 h and 48 h datasets. Values are shown as log-normalized expression centered at 0 (Seurat “scale.data”). (C) Spatial expression of canonical human GBM tumor markers ( CD44 , VIM , TOP2A , and NOTCH1 ) delineating tumor and peri-tumor regions before (0 h) (top) and after (bottom) (48 h) AAV9-hIFNβ treatment. (D) Expression maps of the human IFNβ payload and hallmark IFN-response genes ( CXCL10 , IFIT1 , and IFIT2 ), demonstrating tumor-restricted transgene expression and induction of an IFN-specific transcriptional program within 48 h. (E) Spatial expression of host mouse immune-response genes ( Gfap , Ifitm3 , and Irf7 ) showing localized activation of astroglial and innate immune pathways proximal to the tumor. (F) Integrated datasets (0 and 48 h) visualized using canonical correlation analysis (CCA), showing distinct clustering of tumor and peri-tumor regions (left) and enrichment of IFN-response gene module expression (right). (G) Volcano plot depicting differential gene expression between 0- and 48-h tumor clusters. Red, IFN-response genes; gray, other significantly upregulated genes ( p -Adj <0.01); blue, non-significant. (H) Top enriched Gene Ontology (GO) terms among upregulated genes in 48-h tumor cells, highlighting interferon and inflammatory response pathways (∗∗ p -Adj <0.01; ∗ p -Adj <0.05).

Techniques: Spatial Transcriptomics, Staining, Gene Expression, Marker, Expressing, Activation Assay

Humoral and cellular immunogenicity. (A) Timeline of vaccination and sample collection. (B) The RSV pre-F protein-specific IgG antibody levels of modified LNP-mRNA complexes after the boost immunization, as detected by ELISA. (C) The RSV pre-F protein-specific IgG1 and IgG2a antibody levels and the corresponding ratio of LogIgG2a/LogIgG1 of modified LNP-mRNA complexes after the boost immunization, as detected by ELISA. (D) The neutralizing antibody titers against live virus across all treatment groups. (E) The quantification of restimulated IFN-γ-secreting and IL-17 A-secreting splenocytes was verified in the FluoroSpot test. (F) Results of TEM and TCM in splenic CD4 + T cells. (G) The percentage of naïve CD4 + T cells and CD8 + T cells. (H) Results of TEM and TCM in splenic CD8 + T cells. Data are presented as mean ± SEM. Statistical significance was determined using one-way ANOVA followed by Dunnett's multiple comparisons test, with all comparisons made against the Std-LNP group. All data showed no statistically significant differences and were not labeled in the figure.

Journal: International Journal of Pharmaceutics: X

Article Title: Engineering a safe and potent LNP-mRNA delivery system by leveraging the dual activities of α-tocopherol

doi: 10.1016/j.ijpx.2026.100553

Figure Lengend Snippet: Humoral and cellular immunogenicity. (A) Timeline of vaccination and sample collection. (B) The RSV pre-F protein-specific IgG antibody levels of modified LNP-mRNA complexes after the boost immunization, as detected by ELISA. (C) The RSV pre-F protein-specific IgG1 and IgG2a antibody levels and the corresponding ratio of LogIgG2a/LogIgG1 of modified LNP-mRNA complexes after the boost immunization, as detected by ELISA. (D) The neutralizing antibody titers against live virus across all treatment groups. (E) The quantification of restimulated IFN-γ-secreting and IL-17 A-secreting splenocytes was verified in the FluoroSpot test. (F) Results of TEM and TCM in splenic CD4 + T cells. (G) The percentage of naïve CD4 + T cells and CD8 + T cells. (H) Results of TEM and TCM in splenic CD8 + T cells. Data are presented as mean ± SEM. Statistical significance was determined using one-way ANOVA followed by Dunnett's multiple comparisons test, with all comparisons made against the Std-LNP group. All data showed no statistically significant differences and were not labeled in the figure.

Article Snippet: IFN-γ and IL-17 A FluoroSpot assay: To quantify antigen-specific T cells, splenocytes (2.5 × 10 5 cells/well) were stimulated with an overlapping peptide pool (length = 15 aa, overlap = 10 aa) spanning the RSV pre-F protein (2 μg/mL per peptide) in pre-coated IFN-γ and IL-17 A FluoroSpot plates (Mabtech, Sweden).

Techniques: Immunopeptidomics, Modification, Enzyme-linked Immunosorbent Assay, Virus, Labeling

IFN-γ suppresses tumor growth and invasion. (A) Cytokine profiling of co-culture supernatants via ELISAs: IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α. (B-D) Spatial expression patterns of IFN-γ. (B) Immunofluorescence imaging of the invasive front in SSIT, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green), and grayscale intensity distribution. (C) Immunofluorescence imaging of TIM and NIM, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green). (D) Quantification of relative IFN-γ expression in TIM and NIM. (E) Representative Ki-67 immunohistochemistry images of SSIT cases stratified into IFN-γ-high and IFN-γ-low groups (n=5 each; median split). (F) Quantification of Ki-67 index comparing the two groups. (G) EdU staining demonstrating dose-dependent suppression of TtT/GF pituitary adenoma cell proliferation by IFN-γ (0–100 ng/ml; 48 h). (H) Representative flow cytometry histograms for cell cycle analysis of cells treated with IFN-γ (0–100 ng/ml) in the absence (0 µM) or presence (5 µM) of ruxolitinib. (I) Stacked bar plot showing the percentages of cells in the G 1 , S and G 2 /M phases under the same treatment conditions. (A) One-way ANOVA with Tukey's post hoc multiple comparisons test. (D and F) Unpaired two-tailed Student's t-test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. CTRL, control; DMC, digested mucosal culture; EdU, 5-ethynyl-2′-deoxyuridine; IBA-1, ionised calcium binding adaptor molecule 1; MTC, mucosal tissue culture; NIM, non-invaded mucosa; ns, not significant; PE-A, phycoerythrin-area; SSIT, sphenoid sinus-invasive tumor; TIM, tumor-invaded mucosa.

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: IFN-γ suppresses tumor growth and invasion. (A) Cytokine profiling of co-culture supernatants via ELISAs: IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α. (B-D) Spatial expression patterns of IFN-γ. (B) Immunofluorescence imaging of the invasive front in SSIT, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green), and grayscale intensity distribution. (C) Immunofluorescence imaging of TIM and NIM, showing DAPI (blue), IBA-1 + macrophages (red) and IFN-γ + signals (green). (D) Quantification of relative IFN-γ expression in TIM and NIM. (E) Representative Ki-67 immunohistochemistry images of SSIT cases stratified into IFN-γ-high and IFN-γ-low groups (n=5 each; median split). (F) Quantification of Ki-67 index comparing the two groups. (G) EdU staining demonstrating dose-dependent suppression of TtT/GF pituitary adenoma cell proliferation by IFN-γ (0–100 ng/ml; 48 h). (H) Representative flow cytometry histograms for cell cycle analysis of cells treated with IFN-γ (0–100 ng/ml) in the absence (0 µM) or presence (5 µM) of ruxolitinib. (I) Stacked bar plot showing the percentages of cells in the G 1 , S and G 2 /M phases under the same treatment conditions. (A) One-way ANOVA with Tukey's post hoc multiple comparisons test. (D and F) Unpaired two-tailed Student's t-test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. CTRL, control; DMC, digested mucosal culture; EdU, 5-ethynyl-2′-deoxyuridine; IBA-1, ionised calcium binding adaptor molecule 1; MTC, mucosal tissue culture; NIM, non-invaded mucosa; ns, not significant; PE-A, phycoerythrin-area; SSIT, sphenoid sinus-invasive tumor; TIM, tumor-invaded mucosa.

Article Snippet: Human IFN-γ, IL-1β, IL-6, IL-10, TGF-β and TNF-α levels were quantified using commercial ELISA kits (Human IFN-γ ELISA kit, cat. no. E-EL-H0108; Human IL-1β ELISA kit, cat. no. E-EL-H0149; Human IL-6 ELISA kit, cat. no. E-EL-H0102; Human IL-10 ELISA kit, cat. no. E-EL-H0103; Human TGF-β ELISA kit, cat. no. E-EL-H0110; Human TNF-α ELISA kit, cat. no. E-EL-H0109; Wuhan Elabscience Biotechnology Co., Ltd.).

Techniques: Co-Culture Assay, Expressing, Immunofluorescence, Imaging, Immunohistochemistry, Staining, Flow Cytometry, Cell Cycle Assay, Two Tailed Test, Control, Binding Assay

Elevated IgG levels drive macrophage M2-to-M1 reprogramming. (A) Sphenoid sinus-invasive tumor cases stratified into CD19-high (n=5) and CD19-low (n=5) groups based on the cohort median of CD19 + B cell density, with (B) quantitative analyses of macrophage polarization (M1-like versus M2-like). (C) Dural-invasive tumor and non-invasive tumor cases stratified into IgG-high (n=27) and IgG-low (n=26) groups based on the cohort median of relative IgG immunohistochemistry staining intensity, with (D) quantitative analyses of M1-like/M2-like macrophage proportions. (E and F) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by IgG (10 µg/ml) exposure. Relative (E) IL-6 and (F) TNF-α mRNA expression in RAW264.7 macrophages pre-polarized to M0, M1 or M2 states. (G) Representative flow cytometric cell-cycle profiles of TtT/GF cells following the indicated treatments. (H) Stacked bar plot summarizing the percentages of cells from (G) in G 1 , S and G 2 /M phases. (I) Representative images from the scratch wound assay at 0, 24, 48 and 72 h under the indicated treatments. (J) Quantification of scratch wound closure. (K) Representative western blot images showing total STAT1, p-STAT1, total STAT3, p-STAT3 and β-actin levels in cells treated with IFN-γ (100 ng/ml), IL-6 (100 ng/ml), IFN-γ + IL-6 (50 ng/ml each), ruxolitinib (5 µM) or IFN-γ + IL-6 (50 ng/ml each) plus ruxolitinib (5 µM), as indicated. (L) Densitometric semi-quantification of p-STAT1/STAT1 (ratio). (B and D) Unpaired two-tailed Student's t-test. (E, F, J and L) One-way ANOVA with Tukey's post hoc multiple comparisons test. *P<0.05, ***P<0.001, ****P<0.0001. CTRL, control; IBA-1, ionised calcium binding adaptor molecule 1; ns, not significant; p-, phosphorylated; PE-A, phycoerythrin-area.

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Elevated IgG levels drive macrophage M2-to-M1 reprogramming. (A) Sphenoid sinus-invasive tumor cases stratified into CD19-high (n=5) and CD19-low (n=5) groups based on the cohort median of CD19 + B cell density, with (B) quantitative analyses of macrophage polarization (M1-like versus M2-like). (C) Dural-invasive tumor and non-invasive tumor cases stratified into IgG-high (n=27) and IgG-low (n=26) groups based on the cohort median of relative IgG immunohistochemistry staining intensity, with (D) quantitative analyses of M1-like/M2-like macrophage proportions. (E and F) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by IgG (10 µg/ml) exposure. Relative (E) IL-6 and (F) TNF-α mRNA expression in RAW264.7 macrophages pre-polarized to M0, M1 or M2 states. (G) Representative flow cytometric cell-cycle profiles of TtT/GF cells following the indicated treatments. (H) Stacked bar plot summarizing the percentages of cells from (G) in G 1 , S and G 2 /M phases. (I) Representative images from the scratch wound assay at 0, 24, 48 and 72 h under the indicated treatments. (J) Quantification of scratch wound closure. (K) Representative western blot images showing total STAT1, p-STAT1, total STAT3, p-STAT3 and β-actin levels in cells treated with IFN-γ (100 ng/ml), IL-6 (100 ng/ml), IFN-γ + IL-6 (50 ng/ml each), ruxolitinib (5 µM) or IFN-γ + IL-6 (50 ng/ml each) plus ruxolitinib (5 µM), as indicated. (L) Densitometric semi-quantification of p-STAT1/STAT1 (ratio). (B and D) Unpaired two-tailed Student's t-test. (E, F, J and L) One-way ANOVA with Tukey's post hoc multiple comparisons test. *P<0.05, ***P<0.001, ****P<0.0001. CTRL, control; IBA-1, ionised calcium binding adaptor molecule 1; ns, not significant; p-, phosphorylated; PE-A, phycoerythrin-area.

Techniques: Immunohistochemistry, Staining, Expressing, Scratch Wound Assay Assay, Western Blot, Two Tailed Test, Control, Binding Assay

Anti-CD47 mAb enhances ADCP to suppress tumor cell proliferation. (A) Immunofluorescence staining of CD47 (red) and DAPI (blue) in a representative subset of non-invasive tumor, dural-invasive tumor and sphenoid sinus-invasive tumor cases (n=10 per group). (B) Paired comparison of CD47 fluorescence intensity at the IF versus the TC. (C) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by anti-CD47 mAb (10 µg/ml) treatment for 12 h. Quantitative PCR was used to analyze polarization/activation markers. (D) Schematic illustrating anti-CD47 mAb-mediated blockade of the CD47-SIRPα axis and enhancement of ADCP. (E) EdU assay of TtT/GF cell proliferation in a Transwell co-culture with anti-CD47 mAb-treated polarized macrophages. (F) Quantification of EdU-positive cells. (G) Representative microscopy images and flow cytometry plots showing macrophage phagocytosis of pHrodo™ Red-labeled GFP-TtT/GF cells. (H) Quantification of phagocytosis (%). (B) Paired two-tailed Student's t-test. (C, F and H) One-way ANOVA with Tukey's post hoc multiple comparisons test. **P<0.01, ***P<0.001, ****P<0.0001. ADCP, antibody-dependent cellular phagocytosis; Arg-1, arginase 1; EdU, 5-ethynyl-2′-deoxyuridine; FcγR, Fcγ receptor; GFP, green fluorescent protein; IF, invasive front; mAb, monoclonal antibody; NOS2, nitric oxide synthase 2; ns, not significant; PE, phycoerythrin; SIRPα, signal regulatory protein-α; SSCA, side scatter area; TC, tumor core.

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Anti-CD47 mAb enhances ADCP to suppress tumor cell proliferation. (A) Immunofluorescence staining of CD47 (red) and DAPI (blue) in a representative subset of non-invasive tumor, dural-invasive tumor and sphenoid sinus-invasive tumor cases (n=10 per group). (B) Paired comparison of CD47 fluorescence intensity at the IF versus the TC. (C) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by anti-CD47 mAb (10 µg/ml) treatment for 12 h. Quantitative PCR was used to analyze polarization/activation markers. (D) Schematic illustrating anti-CD47 mAb-mediated blockade of the CD47-SIRPα axis and enhancement of ADCP. (E) EdU assay of TtT/GF cell proliferation in a Transwell co-culture with anti-CD47 mAb-treated polarized macrophages. (F) Quantification of EdU-positive cells. (G) Representative microscopy images and flow cytometry plots showing macrophage phagocytosis of pHrodo™ Red-labeled GFP-TtT/GF cells. (H) Quantification of phagocytosis (%). (B) Paired two-tailed Student's t-test. (C, F and H) One-way ANOVA with Tukey's post hoc multiple comparisons test. **P<0.01, ***P<0.001, ****P<0.0001. ADCP, antibody-dependent cellular phagocytosis; Arg-1, arginase 1; EdU, 5-ethynyl-2′-deoxyuridine; FcγR, Fcγ receptor; GFP, green fluorescent protein; IF, invasive front; mAb, monoclonal antibody; NOS2, nitric oxide synthase 2; ns, not significant; PE, phycoerythrin; SIRPα, signal regulatory protein-α; SSCA, side scatter area; TC, tumor core.

Techniques: Immunofluorescence, Staining, Comparison, Fluorescence, Real-time Polymerase Chain Reaction, Activation Assay, EdU Assay, Co-Culture Assay, Microscopy, Flow Cytometry, Labeling, Two Tailed Test

Summary graphic illustration. This illustration summarizes the proposed model during pituitary adenoma invasion. The tumor invasive front abuts an intact sphenoid sinus mucosa, forming a distinct boundary. The mucosal compartment is enriched for ionised calcium binding adaptor molecule 1-positive macrophages with an M1-like predominance and IgG-high B cells. B cell-derived IgG promotes M2-to-M1 macrophage reprogramming, while coordinated IFN-γ and IL-6 production establishes a tumor-suppressive cytokine gradient that decreases from mucosa toward the tumor core, constraining proliferation and migration via JAK-STAT1 activation. Therapeutically, anti-CD47 monoclonal antibody blocks the CD47-SIRPα ‘don't-eat-me’ axis and augments antibody-dependent cellular phagocytosis, highlighting a strategy for immune checkpoint-targeted therapy that may complement surgical management. FcR, Fc receptor; JAK, Janus kinase; mAb, monoclonal antibody; p-, phosphorylated; SIRPα, signal regulatory protein-α.

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Summary graphic illustration. This illustration summarizes the proposed model during pituitary adenoma invasion. The tumor invasive front abuts an intact sphenoid sinus mucosa, forming a distinct boundary. The mucosal compartment is enriched for ionised calcium binding adaptor molecule 1-positive macrophages with an M1-like predominance and IgG-high B cells. B cell-derived IgG promotes M2-to-M1 macrophage reprogramming, while coordinated IFN-γ and IL-6 production establishes a tumor-suppressive cytokine gradient that decreases from mucosa toward the tumor core, constraining proliferation and migration via JAK-STAT1 activation. Therapeutically, anti-CD47 monoclonal antibody blocks the CD47-SIRPα ‘don't-eat-me’ axis and augments antibody-dependent cellular phagocytosis, highlighting a strategy for immune checkpoint-targeted therapy that may complement surgical management. FcR, Fc receptor; JAK, Janus kinase; mAb, monoclonal antibody; p-, phosphorylated; SIRPα, signal regulatory protein-α.

Techniques: Binding Assay, Derivative Assay, Migration, Activation Assay

Cellular immune responses mapped by peptide pools. A IFN-γ ELISpot responses of splenocytes collected at Day 45 post-prime from BALB/c mice immunized with Arm-Vax, Cl13-Vax, the bivalent Arm+Cl13-Vax, or mock control. Cells were restimulated ex vivo with the indicated GPC peptide pools (Shared, Arm-full, Cl13-full, Arm-only, and Cl13-only), and responses are presented as spot-forming units (SFUs) per 10 6 splenocytes. B Representative ELISpot well images showing responses to a positive control stimulus (PHA), the Shared peptide pool, and a negative control for each immunization group.

Journal: Virus Research

Article Title: Development and protective evaluation of mRNA vaccines encoding the glycoprotein precursor of lymphocytic choriomeningitis virus

doi: 10.1016/j.virusres.2026.199735

Figure Lengend Snippet: Cellular immune responses mapped by peptide pools. A IFN-γ ELISpot responses of splenocytes collected at Day 45 post-prime from BALB/c mice immunized with Arm-Vax, Cl13-Vax, the bivalent Arm+Cl13-Vax, or mock control. Cells were restimulated ex vivo with the indicated GPC peptide pools (Shared, Arm-full, Cl13-full, Arm-only, and Cl13-only), and responses are presented as spot-forming units (SFUs) per 10 6 splenocytes. B Representative ELISpot well images showing responses to a positive control stimulus (PHA), the Shared peptide pool, and a negative control for each immunization group.

Article Snippet: The frequency of antigen-specific IFN-γ-secreting splenocytes was determined using the Mouse IFN-γ Precoated ELISpot Kit (Mabtech, Sweden; Cat# 3321–4HST) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunospot, Control, Ex Vivo, Positive Control, Negative Control

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1) Product Images from "A novel self-amplified RNA vaccine co-expressing NA and HA1 delivered by Salmonella confers potent protection against H9N2 influenza in chickens"

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