Journal: PLoS Pathogens

Article Title: Human plasmacytoid dendritic cells at the crossroad of type I interferon-regulated B cell differentiation and antiviral response to tick-borne encephalitis virus

doi: 10.1371/journal.ppat.1009505

Figure Lengend Snippet: Total PBMC or isolated pDC were left untreated (NS) or stimulated with I-TBEV (dilution 1:12.5) alone or in combination either with two single-chain Ab (scAb) blocking TBEV E glycoprotein, B7 and G7 or with a scAb non-related to TBEV, A10 clone. (A) The release of IFN-α was measured by ELISA in culture supernatants of total PBMC left untreated (NS) or stimulated for 24 hours with I-TBEV alone or in combination either with B7 and G7 or with a non-related scAb A10 clone at the concentration of 0.15 μg/ml, 0.3 μg/ml and 0.6 μg/ml. The results shown were mean values ± standard error of the mean (SEM) of 3 independent experiments. ANOVA p value for IFN-α: 0.006. Based on LSD (equivalent to no adjustments). (B-D) Isolated pDC were left untreated (NS) or stimulated with I-TBEV alone or in combination either with G7 clone (0.6 μg/ml) or with A10 clone (0.6 μg/ml) for 24 hours. The production of IFN-α (B) was measured by ELISA in culture supernatants. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for IFN-α: 0.000. Based on LSD (equivalent to no adjustments). (C) pDC were stained with PD-L1, HLA-DR, CD86, CD80, ILT7, BDCA2 and BDCA4. A total of 50.000 cells were analyzed per sample by flow cytometry to evaluate the percentage of pDC sub-populations. A representative pDC sub-population profile out of 3 different experiments conducted separately is shown: P1-pDC (PD-L1 + CD80 - ) population is indicated in red while P2-pDC (PD-L1 + CD80 + ) in blue. TNF-α production (D) was tested by cytometric bead assay in 24 hour-collected supernatants. The results shown were mean values ± SEM of 3 independent experiments. ANOVA p value for TNF-α: 0.002. Based on LSD (equivalent to no adjustments). (E-F) IgM and IgG production was measured by ELISA in supernatants collected from PBMC after 10 days of stimulation. The results are mean values ± SEM of 3 independent experiments for IgM and 2 independent experiments for IgG. ANOVA p value for IgM: 0.036. Based on LSD (equivalent to no adjustments).

Article Snippet: Single-chain Fragment variable–Fragment crystallizable antibodies (scAb) specific for TBEV E glycoprotein, B7 and G7 clones, or non-specific as A10 clone were generated by Yumab (Braunschweig, Germany) and provided by GSK.

Techniques: Isolation, Blocking Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Staining, Flow Cytometry

Journal: Diagnostics

Article Title: Reference Intervals for Common Biochemistry and Hematology Parameters in Early Pregnancy—A Prospective Study

doi: 10.3390/diagnostics15040415

Figure Lengend Snippet: The analytical methods used in the study and their analytical performance assessed according to the specifications of the Biological variation database [ 9 ], with the exception of the erythrocyte sedimentation rate (ESR), which was assessed according to Penev M et al. [ 10 ]. A complete blood count was performed using the Beckman Coulter DxH800 (Beckman Coulter, Brea, CA, USA), the erythrocyte sedimentation rate was assessed using the iSED (Alcor Scientific, Smithfield, RI, USA), and all biochemistry tests were determined on the Roche Cobas 6000 c501 (Roche diagnostics, Mannheim, Germany), except for ferritin and HDL cholesterol which were analyzed using the Beckman Coulter AU480 (Beckman Coulter, Brea, CA, USA).

Article Snippet: Alpha amylase (AMY) (U/L) , IFCC (enzymatic with E-G 7 PNP) , 1.7 , 4.9 , 3.3 , 1.6.

Techniques: Sedimentation, Cell Counting, Derivative Assay, Concentration Assay, Immunoturbidimetry Assay, Colorimetric Assay, Binding Assay, Iron Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: HFB301001, an OX40-based immunotherapy, drives Treg clearance and CTL activation through optimized OX40 receptor clustering

doi: 10.1136/jitc-2025-014185

Figure Lengend Snippet: High-affinity mutants of HFB301001 exhibited inferior antitumor efficacy than HFB301001 in vivo. ( A ) Schematic representation of HFB301001 and HFB301001 hi antitumor efficacy evaluation. MC38 or E.G7 tumor-bearing mice were treated with HFB301001 or HFB301001 hi for four doses, and tumor volumes were measured every 3 days. ( B, E ) Individual tumor growth curves, ( C, D ) survival curves, and ( F, G ) body weights were monitored (n=10). Statistical significance of survival was determined using the log-rank test. Data in (C) and (D) are presented as mean±SEM from one representative experiment of two independent replicates. ***p<0.001, ****p<0.0001.

Article Snippet: The mouse colon cancer MC38 and E.G7 cell lines were purchased from Shanghai Model Organisms Center.

Techniques: In Vivo

Journal: Journal for Immunotherapy of Cancer

Article Title: HFB301001, an OX40-based immunotherapy, drives Treg clearance and CTL activation through optimized OX40 receptor clustering

doi: 10.1136/jitc-2025-014185

Figure Lengend Snippet: HFB301001 promotes robust CD8 + T cell activation and establishes immunological memory. ( A ) The antitumor efficacy of HFB301001 at different doses. MC38 tumor-bearing hOX40 mice were treated with HFB301001 at various doses (n=10). Survival curve was plotted. Statistical significance of survival was determined using the log-rank test. ( B ) Schematic illustration of the rechallenge assay. MC38 tumor-bearing mice cured by HFB301001 and age-matched naïve controls were rechallenged with MC38 cells 2 months after the initial tumor clearance. Survival curve was subsequently monitored. The rechallenged mice (n=6) represent animals accumulated from multiple independent experimental cohorts, rather than a single experiment. ( C ) CD4 + and CD8 + T cell depletion assay. MC38 tumor-bearing mice were treated with HFB301001 or HFB301001 hi for three doses. Depleting antibodies against CD4 + or CD8 + T cells were administered intraperitoneally (i.p.) 1 day before treatment. Tumor volume was monitored. ( D ) UMAP plot of lymphoid cells. MC38 tumor-bearing mice were treated with PBS or HFB301001 for three doses. Tumor-infiltrating immune cells were analyzed by scRNA-seq and colored by T-cell subclusters. ( E ) Proportions of T-cell subclusters in control (PBS) and HFB301001-treated groups. ( F ) Bubble diagram depicting the expression patterns of activation and exhaustion across distinct T cell clusters. ( G ) GO-enriched functional pathways of tumor-infiltrating Treg, CD4 + T, and CD8 + T cells in PBS (Control) vs HFB301001-treated mice. Colors: Treg (purple), CD4 + (green), CD8 + (orange). ( H–J ) Schematic illustration of immunofluorescence analysis of mouse tumor tissues. MC38 tumor-bearing mice were treated with HFB301001 or HFB301001 hi for three doses. Tumor tissues were harvested and subjected to multicolor immunofluorescence staining. Blue indicates nuclei, red indicates CD4 or CD8, and green indicates Foxp3. ( K ) Schematic illustration of T cell-specific activation. E.G7-OVA tumor-bearing mice were treated with HFB301001 for three doses. Splenocytes were harvested and incubated with E.G7-OVA cells for 36 hours, after which IFN-γ production was measured. ( L ) The representative wells (left) and the number of IFN-γ + spots per 2×10⁴ cells (right) are shown. Data are presented as means±SD. n=3 for (L), n=6–10 for A–C), p values were calculated using the t-test in (L) and two-way ANOVA in (C). *p<0.05, ****p<0.0001. ANOVA, analysis of variance; ns, not significant; PBS, Phosphate-buffered saline; scRNA-seq, single-cell RNA sequencing; UMAP, Uniform Manifold Approximation and Projection.

Article Snippet: The mouse colon cancer MC38 and E.G7 cell lines were purchased from Shanghai Model Organisms Center.

Techniques: Activation Assay, Depletion Assay, Control, Expressing, Functional Assay, Immunofluorescence, Multicolor Immunofluorescence Staining, Incubation, Saline, Single Cell, RNA Sequencing