Journal: Infection and Immunity

Article Title: Comparison of Pathogenesis and Host Immune Responses to Candida glabrata and Candida albicans in Systemically Infected Immunocompetent Mice

doi: 10.1128/iai.69.8.5046-5055.2001

Figure Lengend Snippet: FIG. 4. Temporal expression of TNF-a, IL-12, IFN-g, and IL-10 proteins in kidneys during C. glabrata and C. albicans infection. Crl:CF-1 mice were infected with virulent C. glabrata or C. albicans as described for Fig. 3. At specific time points p.i., the mice were euthanized, and kidneys were excised and homogenized. Levels of the immunoreactive cytokines TNF-a (a), IL-12 p70 (b), IFN-g (c), and IL-10 (d) were quantified in kidney homogenates. Results represent the mean 6 SEM of two separate experiments, 8 to 14 mice per treatment group. p, significantly greater than control mice, P , 0.05.

Article Snippet: Protein levels of TNF-a, IL-12, IL-10, and IFN-g were subsequently measured using commercially available cytokinespecific murine ELISA kits (Quantikine mouse TNF-a, mouse IL-12 p70, mouse IFN-g, and mouse IL-10; R&D systems, Minneapolis, Minn.) according to the manufacturer’s directions.

Techniques: Expressing, Infection, Control

Journal: Frontiers in Immunology

Article Title: NECA alleviates inflammatory responses in diabetic retinopathy through dendritic cell toll-like receptor signaling pathway

doi: 10.3389/fimmu.2024.1415004

Figure Lengend Snippet: NECA influenced the presence of DC surface markers, release of inflammatory cytokines and mRNA/protein expression in TLR pathway in healthy individuals. (A) Histograms were overlapped to compare NECA treated DCs (n = 10) with controls (n = 10) from the representative experiment, and the variation in MFI for surface markers of CD40, CD80, CD86, HLA-DR. (B) IL-6, IL-10, IL-12/p70 and TNF-2 were measured by ELISA in NECA-treated DCs (n = 10) vs. vehicles (n = 10). (C, D) MRNA relative transcription level and protein expression level of TLR-4, MYD88, and NF-88 in DCs of NECA treated (n = 10) vs. vehicles (n = 10) by PCR and WB. Statistical analysis involved the utilization of the paired samples t-test. (**P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: The levels of IL-12 p70 was measured using mouse IL-12 p70 multi-set enzymic immunosorbent assay kit (R&D Systems) and human IL-12 p70 high-sensitivity ELISA kit (eBioscience).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: NECA alleviates inflammatory responses in diabetic retinopathy through dendritic cell toll-like receptor signaling pathway

doi: 10.3389/fimmu.2024.1415004

Figure Lengend Snippet: NECA influenced the presence of DC surface markers, release of inflammatory cytokines and mRNA/protein expression in TLR pathway in patients with DM. (A) Histograms were overlapped to compare NECA treated DCs (n = 10) with controls (n = 10) from the representative experiment, and the variation in MFI for surface markers of CD40, CD80, CD86, HLA-DR. (B) IL-6, IL-10, IL-12/p70 and TNF-2 were measured by ELISA in NECA-treated DCs (n = 10) vs. vehicles (n = 10). (C, D) MRNA relative transcription level and protein expression level of TLR-4, MYD88, and NF-88 in DCs of NECA treated (n = 10) vs. vehicles (n = 10) by PCR and WB. Statistical analysis involved the utilization of the paired samples t-test. (ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: The levels of IL-12 p70 was measured using mouse IL-12 p70 multi-set enzymic immunosorbent assay kit (R&D Systems) and human IL-12 p70 high-sensitivity ELISA kit (eBioscience).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: NECA alleviates inflammatory responses in diabetic retinopathy through dendritic cell toll-like receptor signaling pathway

doi: 10.3389/fimmu.2024.1415004

Figure Lengend Snippet: NECA influenced the presence of BMDC surface markers, release of inflammatory cytokines and mRNA/protein expression in TLR pathway in mice. (A) Histograms were overlapped to compare NECA treated BMDCs (n = 4) with controls (n = 4) from the representative experiment, and the variation in MFI for surface markers of CD40, CD80, CD86, MHCII. (B) IL-6, IL-10, IL-12/p70 and TNF-2 were measured by ELISA in NECA-treated BMDCs (n = 4) vs. vehicles (n = 4). (C, D) MRNA relative transcription level and protein expression level of TLR-4, MYD88, and NF-88 in BMDCs of NECA-treated (n = 4) vs. vehicles (n = 4) by PCR and WB. Statistical analysis involved the utilization of the paired samples t-test and Wilcoxon. (ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: The levels of IL-12 p70 was measured using mouse IL-12 p70 multi-set enzymic immunosorbent assay kit (R&D Systems) and human IL-12 p70 high-sensitivity ELISA kit (eBioscience).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: NECA alleviates inflammatory responses in diabetic retinopathy through dendritic cell toll-like receptor signaling pathway

doi: 10.3389/fimmu.2024.1415004

Figure Lengend Snippet: NECA influenced the presence of DC surface markers, release of inflammatory cytokines and mRNA/protein expression in TLR pathway in patients with DR. (A) Histograms were overlapped to compare NECA treated DCs (n = 10) with controls (n = 10) from the representative experiment, and the variation in MFI for surface markers of CD40, CD80, CD86, HLA-DR. (B) IL-6, IL-10, IL-12/p70 and TNF-2 were measured by ELISA in NECA-treated DCs (n = 10) vs. vehicles (n = 10). (C, D) MRNA relative transcription level and protein expression level of TLR-4, MYD88, and NF-88 in DCs of NECA-treated (n = 10) vs. vehicles (n = 10) by PCR and WB. Statistical analysis involved the utilization of the paired samples t-test and Wilcoxon test. (ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001,****P < 0.0001).

Article Snippet: The levels of IL-12 p70 was measured using mouse IL-12 p70 multi-set enzymic immunosorbent assay kit (R&D Systems) and human IL-12 p70 high-sensitivity ELISA kit (eBioscience).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: NECA alleviates inflammatory responses in diabetic retinopathy through dendritic cell toll-like receptor signaling pathway

doi: 10.3389/fimmu.2024.1415004

Figure Lengend Snippet: MRS1754 inhibited the effect of NECA on DC surface markers and release of inflammatory cytokines in healthy individuals. (A) Histograms were overlapped to compare NECA treated DCs (n = 10) with NECA+MRS1754-treated DCs (n = 10) from the representative experiment, and the variation in MFI for surface markers of CD40, CD80, CD86, HLA-DR. (B) IL-6, IL-10, IL-12/p70 and TNF-2 were measured by ELISA in NECA-treated DCs (n = 8) vs. NECA+MRS1754-treated DCs (n = 8). Statistical analysis involved the utilization of the paired samples t-test and Wilcoxon test. (ns: not significant, *P < 0.05, **P < 0.01).

Article Snippet: The levels of IL-12 p70 was measured using mouse IL-12 p70 multi-set enzymic immunosorbent assay kit (R&D Systems) and human IL-12 p70 high-sensitivity ELISA kit (eBioscience).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: NECA alleviates inflammatory responses in diabetic retinopathy through dendritic cell toll-like receptor signaling pathway

doi: 10.3389/fimmu.2024.1415004

Figure Lengend Snippet: Effect of NECA on BMDCs function in vivo in mice. (A) Histograms were overlapped to compare NECA treated BMDCs (n = 4) with controls (n = 4) from the representative experiment, and the variation in MFI for surface markers of CD40, CD80, CD86, MHCII in EDR mice. (B) IL-6, IL-10, IL-12/p70 and TNF-2 were measured by ELISA in NECA-treated BMDCs (n = 4) vs. vehicles (n = 4). (C, D) MRNA relative transcription level and protein expression level of TLR-4, MYD88, and NF-88 in BMDCs of NECA-treated (n = 4) vs. vehicles (n = 4) by PCR and WB. Statistical analysis involved the utilization of the paired samples t-test. (ns: not significant, **P < 0.01, ***P < 0.001, ****P < 0.0001).

Article Snippet: The levels of IL-12 p70 was measured using mouse IL-12 p70 multi-set enzymic immunosorbent assay kit (R&D Systems) and human IL-12 p70 high-sensitivity ELISA kit (eBioscience).

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Expressing

Journal: International Journal of Nanomedicine

Article Title: IL-12-Overexpressed Nanoparticles Suppress the Proliferation of Melanoma Through Inducing ICD and Activating DC, CD8 + T, and CD4 + T Cells

doi: 10.2147/IJN.S442446

Figure Lengend Snippet: CPP transferred IL-12 into B16F10 cells and CPP-IL-12 induced immunogenic cell death. ( A ) The structure of IL-12 vector. ( B ) Western blot detection of IL-12 expression. ** P < 0.01, Student’s t -test. ( C ) ELISA. CPP-IL-12 increased IL-12 levels in the supernatant of B16-F10 cells, and 2.0 W/cm 2 of laser further increased IL-12 levels. ** P < 0.01, ANOVA. ( D ) Live/dead staining assay. Scale bars=100 μm. ( E ) The number of dead cells. CPP-IL-12 increased and 2.0 W/cm 2 of laser further increased the number of dead cells. ** P < 0.01, ANOVA. ( F ) Cell apoptosis analysis. CPP-IL-12 increased and laser treatment further increased the number of apoptotic cells. ( G ) Immunofluorescence. Green color, CALR expression. Scale bars=2 μm. ( H, I ) ELISA analysis of extracellular and intracellular HMGB1 levels, respectively. * P < 0.05, ** P < 0.01, ANOVA. (J) ATP levels. The ATP levels decreased in CPP-IL-12-treated cells, and laser treatment further reduced ATP content. ** P < 0.01, ANOVA. ( K ) CPP increased CD80 + /CD86 + expression of DC.

Article Snippet: IL-12 and HMGB1 levels were estimated using ELISA kits (CSB-E04600m and CSB-E08225M, CUSABIO, Wuhan, China) in accordance with the manufacturer′s instructions.

Techniques: Plasmid Preparation, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Staining, Immunofluorescence