Journal: Frontiers in Public Health

Article Title: Proton beam therapy induces protective immunity via HMGB1-dependent signaling

doi: 10.3389/fpubh.2025.1686678

Figure Lengend Snippet: Proton radiation stimulates immunogenic cell death. (A) Experimental schedule for proton-induced ICD evaluation. Primary tumor growth (B) and distal tumor growth (C) after the transplantation of different doses (3, 6, 12 Gy) proton-irradiated cells. Four weeks after tumor cell transplantation, spleens were harvested and analysed: immunohistochemical staining of T lymphocytes (D) and IFN- γ (G) infiltration in the spleen, and quantification by flow cytometry (E,F) . (H) The expression of HMGB1 in serum after the transplantation of proton-irradiated cells. * means p < 0.05, ** means p < 0.01,*** means p < 0.001, **** means p < 0.0001, ns means not significant.

Article Snippet: HMGB1 and Interferon-gamma (IFN-γ) in the mouse serum samples were measured by using HMGB1 kit (H257-1-2, Mouse HMGB1 ELISA Kit, NJJC Bioscience, Nanjing, China) and IFN-γ kit (#EK0375, Boster Biological Technology, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Transplantation Assay, Irradiation, Immunohistochemical staining, Staining, Flow Cytometry, Expressing

Journal: Frontiers in tropical diseases

Article Title: Adjuvants Differentially Modulate the Immunogenicity of Lassa Virus Glycoprotein Subunits in Mice

doi: 10.3389/fitd.2022.847598

Figure Lengend Snippet: Cytokine secretion measured from mouse splenocytes using a FluoroSpot assay. Geometric mean (GMT) with 95% confidence intervals of (A) IL-2, (B) TNF-α, (C) IFN-γ and (D) polyfunctional cytokine secretion from splenocytes harvested from mouse groups one-week post-dose 2 and stimulated with a LASV GP peptide pool for 24 hours. (n=3 for the ISA 720 groups, n=1-2 for LiteVax groups, and n=3-4 for GPI-0100 groups) A two-way ANOVA, followed by a Tukey’s multiple comparison was used to compare the number of cytokine-secreting cells from immunized groups to the group receiving LASV GP alone, and between different adjuvant formulations of the same antigen concentration, respectively. (*p < 0.05, **p < 0.01).

Article Snippet: The FluoroSpot assay was performed using mouse IFN-γ/TNF-α/IL-2 Triple-Color FluoroSpot kits (Cellular Technology Limited (CTL), Cleveland, OH).

Techniques: Flurospot, Comparison, Adjuvant, Concentration Assay

Journal: Frontiers in Immunology

Article Title: IL-15 Harnesses Pro-inflammatory Function of TEMRA CD8 in Kidney-Transplant Recipients

doi: 10.3389/fimmu.2017.00778

Figure Lengend Snippet: IL-15-stimulated TEMRA CD8 from Kidney-Transplant (KT) Recipients induces the activation of endothelium through IFN-γ and TNF-α secretion. (A) Expression of CD25 or CD69 by CD8 T cell subsets purified from KT recipients stimulated for 24 h with plate-bound anti-CD3 (2 µg/mL) and IL-15 (10 ng/mL). Representative flow data are shown, and bars indicate mean ± SEM of pooled data ( n = 10–12). (B) Schematic overview of the strategy to assess the inflammation of human umbilical vein endothelial cell (HUVEC) by IL-15-activated CD8 T cell subsets from KT. (C) Expression of CX3CL1 by HUVEC after 6 h of culture in the presence of medium control or selective inhibitors [anti-IFN-γ (10 µg/mL) or anti-TNF-γ (10 µg/mL) mAb] and with supernatant (dilution of supernatant of 1/6) from CD8 T cell subsets from KT stimulated overnight with plate-bound anti-CD3 (2 µg/mL) and IL-15 (10 ng/mL). Bars indicate mean ± SEM of pooled data ( n = 4–6). Mann–Whitney test (A) or Kruskal–Wallis test followed by a Dunn’s Multiple Comparison Test (C) was performed (* p < 0.05, ** p < 0.01, **** p < 0.0001).

Article Snippet: Antibodies against the following proteins were purchased from BD Biosciences: CD3 (HIT3a and UCHT1), CD69 (SK7), CD25 (M-A251), CD28 (CD28.2), pSTAT5 pY694 (clone 47), p38MAPK pT180/pY182 (38/p38), TNFα (Mab11), IFN-γ (B27), and Annexin V. Antibodies against the following proteins were purchased from Miltenyi Biotech: CD8 (BW135/80), CD45RA (T6D11), CCR7 (REA108), pAKT pS473 (REA359), and pERK1/2 pT202/pY204 (REA152).

Techniques: Activation Assay, Expressing, Purification, Control, MANN-WHITNEY, Comparison