Journal: Frontiers in Immunology

Article Title: Engineered antibody cytokine chimera synergizes with DNA-launched nanoparticle vaccines to potentiate melanoma suppression in vivo

doi: 10.3389/fimmu.2023.1072810

Figure Lengend Snippet: Design, pharmacokinetic and pharmacodynamic profiles of novel ACC TA99-HL2-KOA1. (A) Model depiction of engineered cytokine mimic HL2-KOA1 binding to IL-2Rα. IL-2 shown in blue, IL-2Rα shown in grey. Positions in contact with F42 shown as grey spheres. F42 and F42V shown as pink and red spheres respectively. Clash scores provided in total Rosetta energy units (REU). (B) Reducing SDS-PAGE analysis comparing the migration patterns of TA99-WT, TA99-Neo2/15, TA99-HL2-KOA1, pVAX backbone control transfection supernatants. (C) Binding of recombinant TA99-WT, TA99-Neo2/15, and TA99-HL2-KOA1 to recombinant human Tyrp1 compared to a murine isotype IgG2a control. (D) Binding of recombinant TA99-WT, TA99-Neo2/15, and TA99-HL2-KOA1 to recombinant human IL2-Rα compared to human IL2-Fc control. (E) Binding of recombinant TA99-WT, TA99-Neo2/15, and TA99-HL2-KOA1 to recombinant human IL2-Rβ compared to human IL2-Fc control. (F) Geometric mean fluorescence intensity of phosphorylated STAT5 in CD25+ CD4+ CD19- T cells from naïve mice (n=6) treated with indicated ACC. (G) Pharmacokinetic profile of recombinant TA99-WT and TA99-Neo-2/15 antibodies; C57BL/6 mice were injected intraperitoneally with 100µg of ACC constructs (n=5 mice per group). Serum at the indicated timepoints was assessed for TYRP1 binding by ELISA. (H) Total serum cytokine level over 14-day period in terms of area under the curve (AUC) in mice treated with ACC. (I) Timecourse of serum TNFα levels in sera of mice administered 100µg ACC. Two sera pools were used for (H, I) . (J) Survival curves of mice (n=10 mice per group) post tumor challenge following treatment with anti-PD1 alone, or anti-PD1 (200µg) + TA99-Neo2/15 (50µg) + TriVax (10µg each of Trp2, Gp100 and Tyrp1), or anti-PD1 (200µg) + TA99-HL2-KOA1 (50µg) + TriVax. 1x10 5 B16F10 cells subcutaneously into the right flank of C57BL/6 mice on Day 0. Green arrows indicate administration of treatment. (K) Survival curves of mice (n=10 mice per group) post tumor challenge following treatment with anti-PD1 alone, or anti-PD1 (200µg) + TA99-HL2-KOA1 (50µg) + TriVax, or anti-PD1 + TA99 (50µg) + Human IL2 (5µg) + TriVax. Challenge and treatment scheme was performed as in (J) Error bars represent standard deviation; non-parametric Mann Whitney T test compared with TA99-WT used in (F) ; non-parametric Kruskal-Wallis ANOVA was used in (I) ; log-rank test was used to compare between differences in all survival curves; *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: In addition, serially diluted recombinant protein variants of antibody-cytokine chimera or recombinant mouse Fc-tagged IL2 (IL2-Fc) (Molecular Innovations, Cat# MIL2-FC-0.05MG) were used in place of serially diluted mouse sera.

Techniques: Binding Assay, SDS Page, Migration, Control, Transfection, Recombinant, Fluorescence, Injection, Construct, Enzyme-linked Immunosorbent Assay, Standard Deviation, MANN-WHITNEY

Journal: BMC Cancer

Article Title: HLA class II-restricted T cell epitopes in public neoantigens of ESR1 and PIK3CA in breast cancer

doi: 10.1186/s12885-025-13992-6

Figure Lengend Snippet: Summary of the ELISPOT assay of healthy donors. A Representative data of the ELISPOT assay for ES#2_E380Q, PIK#5_E545A, PIK#8_H1047L, and PIK#9_H1047Y, measured by IFN-γ (red spots) and IL-2 (blue spots). The numbers denote the spot count for IFN-γ (red). NC, negative control. TNTC: too numerous to count. B , C The ELISPOT profiles for DRB4*01:03 positive donors ( B ) and DP5 positive donors ( C ). The data for peptide pairs that showed penetrance ≥ 0.4 and g15 ratio > 2.0 are presented. HLA alleles of each donor are displayed on the top. Positive ELISPOT responses (IFN-γ) are highlighted in red. Numbers indicate the positive responses/total number of experiments (range of spot counts). The penetrance represents the number of positive donors/numbers of total donors (in red letters). The peptides with %Rank < 10 (NetMHCIIpan-4.1) is highlighted in yellow, and those with a g15 ratio > 2.0 (MHC-density assay) is highlighted in light blue. NA: not analyzed (shadowed in gray). TNTC: too numerous to count

Article Snippet: The ELISPOT assay was conducted using the human interferon-gamma (IFN-γ)/IL-2 double-color ELISPOT kit (Cellular Technology Limited, Cleveland, OH, USA), following the manufacturer’s instructions.

Techniques: Enzyme-linked Immunospot, Negative Control