Journal: Communications Biology

Article Title: Tick saliva reprograms macrophages into immunosuppressive hubs that regulate T-cell immunity in Rhipicephalus microplus infestation

doi: 10.1038/s42003-026-09981-5

Figure Lengend Snippet: A Schematic overview of the experimental design for T cell response analysis. Concentrations of TNF ( B ) and IFN-γ ( C ) in culture supernatants of PBMCs or CD14⁻ PBMCs stimulated with anti-CD3/CD28 antibodies or ConA stimulation in the presence or absence of Rm-saliva, measured by ELISA. D Representative flow cytometry plots showing intracellular TNF and IFN-γ expression in CD4⁺ T cells under the same stimulation conditions. Frequencies of TNF⁺ ( E ), IFN-γ⁺ ( F ), and TNF⁺IFN-γ⁺ ( G ) T cells in PBMCs or CD14⁻ PBMCs following anti-CD3/CD28 and Rm-saliva stimulation. H Representative plots for CD8⁺ T cells under identical conditions. Corresponding frequencies of TNF⁺ ( I ), IFN-γ⁺ ( J ), and TNF⁺IFN-γ⁺ ( K ) CD8⁺ T cells. L Representative flow cytometry plots showing intracellular granzyme B and perforin expression in CD8⁺ T cells under the same stimulation conditions. Frequencies of granzyme B⁺ ( M ), perforin⁺ ( N ), and granzyme B⁺perforin⁺ ( O ) CD8⁺ T cells in PBMCs or CD14⁻ PBMCs following anti-CD3/CD28 and Rm-saliva stimulation. Data represent paired samples from nine individual cattle ( n = 9). * P < 0.05 (Friedman test followed by the Wilcoxon signed-rank test). Illustration elements in this figure are from the NIAID NIH BioArt Source (bioart.niaid.nih.gov/bioart/509).

Article Snippet: IFN-γ and TNF were quantified using the Bovine IFN-γ ELISA Development Kit (Mabtech, Nacka Strand, Sweden) and the ELISA Flex: Bovine TNF-α (Mabtech).

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing

Journal: Communications Biology

Article Title: Tick saliva reprograms macrophages into immunosuppressive hubs that regulate T-cell immunity in Rhipicephalus microplus infestation

doi: 10.1038/s42003-026-09981-5

Figure Lengend Snippet: A IL-10 concentration in culture supernatants of PBMCs or CD14⁻ PBMCs stimulated with anti-CD3/CD28 antibodies in the presence or absence of Rm-saliva, measured by ELISA. B Representative flow cytometry plots showing intracellular IL-10 and TGF-β expression in CD4⁺ T cells under the same stimulation conditions. Frequencies of IL-10⁺ ( C ), TGF-β⁺ ( D ), and IL-10⁺TGF-β⁺ double-positive ( E ) CD4⁺ T cells in PBMCs or CD14⁻ PBMCs following anti-CD3/CD28 and Rm-saliva stimulation. F Summary of the proportions of IL-10⁺, TGF-β⁺, and IL-10⁺TGF-β⁺ CD4⁺ T cells. Data represent paired samples from nine individual cattle ( n = 9). * P < 0.05 (Friedman test followed by the Wilcoxon signed-rank test). G Representative flow cytometry plots showing the proportion of CD25⁺Foxp3⁺ cells among CD4⁺ T cells in the presence or absence of Rm-saliva, with or without CD14⁺ cells. H Summary of the frequency of CD25⁺Foxp3⁺ Tregs among CD4⁺ T cells under the same conditions. I Comparison of IL-10⁺ cell distribution between conventional CD4⁺ T cells (Tconv) and Tregs, with or without CD14⁺ cells. J Representative flow cytometry plots showing intracellular IL-10 and TGF-β expression in Tregs. Frequencies of IL-10⁺ ( K ), TGF-β⁺ ( L ), and IL-10⁺TGF-β⁺ ( M ) cells within Tregs. N Summary of the proportions of IL-10⁺, TGF-β⁺, and IL-10⁺TGF-β⁺ Tregs. Data represent paired samples from nine individual cattle ( n = 9). * P < 0.05 (Friedman test followed by the Wilcoxon signed-rank test).

Article Snippet: IFN-γ and TNF were quantified using the Bovine IFN-γ ELISA Development Kit (Mabtech, Nacka Strand, Sweden) and the ELISA Flex: Bovine TNF-α (Mabtech).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Comparison

Journal: Communications Biology

Article Title: Tick saliva reprograms macrophages into immunosuppressive hubs that regulate T-cell immunity in Rhipicephalus microplus infestation

doi: 10.1038/s42003-026-09981-5

Figure Lengend Snippet: Concentrations of TNF ( A ), IL-1β ( B ), and IL-10 ( C ) in culture supernatants of macrophages stimulated with LPS and IFN-γ in the presence or absence of Rm-saliva, measured by ELISA. D – I Flow cytometry analysis of M1 and M2 markers in macrophages. D Representative plots of CD80 and TNF co-expression. E Summary of the frequency of CD80⁺TNF⁺ macrophages. F Representative plots of CD206 and IL-10 expression. G Summary of the frequency of CD206⁺IL-10⁺ macrophages. H Representative plots of CD206 and TGF-β expression. I Summary of the frequency of CD206⁺TGF-β⁺ macrophages. Ratio of M1-like (CD80⁺TNF⁺) to M2-like (CD206⁺IL-10⁺ or CD206⁺TGF-β⁺) macrophages in individual samples ( J ) and the overall proportion of M1/M2 phenotypes ( K ). Data represent paired samples from nine individual cattle ( n = 9). * P < 0.05 (Friedman test followed by the Wilcoxon signed-rank test).

Article Snippet: IFN-γ and TNF were quantified using the Bovine IFN-γ ELISA Development Kit (Mabtech, Nacka Strand, Sweden) and the ELISA Flex: Bovine TNF-α (Mabtech).

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing

Journal: Communications Biology

Article Title: Tick saliva reprograms macrophages into immunosuppressive hubs that regulate T-cell immunity in Rhipicephalus microplus infestation

doi: 10.1038/s42003-026-09981-5

Figure Lengend Snippet: A Representative histograms showing the expression intensity of MHC class II on macrophages stimulated with LPS and IFN-γ in the presence or absence of Rm-saliva. B Summary of the MFI of MHC class II on macrophages. C Summary of the proportions of MHC class II⁺ on macrophages. D Representative flow cytometry plots showing co-expression of MHC class II and CD80. E Summary of the frequencies of MHC class II⁺CD80⁺ macrophages. F Representative flow cytometry plots showing co-expression of MHC class II and CD86. G Summary of the frequencies of MHC class II⁺CD86⁺ macrophages. Concentrations of CCL2 ( H ), CCL3 ( I ), CCL4 ( J ), CCL5 ( K ), CCL8 ( L ), CXCL9 (M) in culture supernatants of macrophages stimulated with LPS and IFN-γ in the presence or absence of Rm-saliva. Chemokine concentrations were generally low under unstimulated conditions, with some values falling below the assay’s detection limit. Data represent paired samples from nine individual cattle ( n = 9). * P < 0.05 (Friedman test followed by the Wilcoxon signed-rank test).

Article Snippet: IFN-γ and TNF were quantified using the Bovine IFN-γ ELISA Development Kit (Mabtech, Nacka Strand, Sweden) and the ELISA Flex: Bovine TNF-α (Mabtech).

Techniques: Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: Multifaceted immune resistance landscapes in human oligodendrocytes protect against cytotoxic T cells and are dysregulated in MS brain cell subsets

doi: 10.64898/2026.04.21.719872

Figure Lengend Snippet: (A) FACS analysis of antigen specificity of FluTC generated from HLA-A2 + healthy donors by repetitive expansions. All samples were gated on lymphocytes, single cells, and live cells. CD8 and Flu-Pentamer stainings were performed on days 7 and 14 of antigen-specific expansion (ASE) and day 14 of rapid expansion protocol (REP). During ASE, some CD8 + T cells were expanded in the presence of unpulsed feeder cells (w/o flu-peptide) as a negative control. (B) ELISA to determine cytokine secretion by FluTC. MO3.13-A2-Luc cells were pulsed with diluting concentrations of flu peptide and co-cultured with FluTC. After 20 h of co-culture, the supernatant of co-culture was analyzed by ELISA to detect IFNγ (left) and Granzyme B (right) secretion by FluTC.

Article Snippet: Supernatants of FluTC-MO3.13-A2-Luc co-cultures and anti-CD3/CD28 activated FluTC were further analyzed for the detection of IFNγ (Human IFN-γ ELISA Set, BD OptEIA, #555142), TNFα (Human TNF ELISA Set, BD OptEIA, #555212), Granzyme B (Human Granzyme B ELISA development kit, Mabtech, #3485-1H-20) and TRAIL (human TRAIL/TNFSF10 DuoSet ELISA, R&D Systems, #DY375-05) ( , ).

Techniques: Generated, Negative Control, Enzyme-linked Immunosorbent Assay, Cell Culture, Co-Culture Assay

Journal: Parasite Immunology

Article Title: The Regulatory Effect of Histone Deacetylase ( HDAC ) 1 and 2 on iNOS , IL ‐6, TNF ‐α and IL ‐10 Expression in Canine Macrophages Infected With Leishmania infantum

doi: 10.1111/pim.70078

Figure Lengend Snippet: Inhibition of HDAC1 in cells infected with Leishmania infantum and treated with sodium butyrate on IL‐6, TNF‐α and IL‐10 expression. DH82 cells were treated with the pharmacological HDAC1 inhibitor sodium butyrate NaB (10 mM) and (20 mM). After 6 h of infection, the culture supernatant cells were harvested, and (A) IL‐6, (B) TNF‐α and (C) IL‐10 were measured by capture ELISA. The statistical test used was the Friedman test, followed by Dunn's multiple‐comparison test ( p < 0.05).

Article Snippet: Cytokines IL‐6, TNF‐α and IL‐10 were quantified in culture supernatant from L. infantum infected macrophages after HDAC1 and HDAC2 suppression with NaB (10 mM) and (20 mM) (Sigma‐Aldrich, MO, USA) by using the DuoSet ELISA Development Systems canine kits (R&D Systems, Minneapolis, MN, USA), following the manufacturer's instructions.

Techniques: Inhibition, Infection, Expressing, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Veterinary Sciences

Article Title: In Vitro Characterization of an Equinized Anti-PD-L1 Antibody for Cancer Immunotherapy in Horses

doi: 10.3390/vetsci13040343

Figure Lengend Snippet: Binding inhibition of equine PD-1/PD-L1 and enhancement of cytokine production and cell proliferation by Eq6C11 treatment. ( a ) The blocking activity of anti-PD-L1 antibodies on the binding of EqPD-L1-Ig to plate-coated EqPD-1-Ig. Equine IgG was used as negative control antibody. The relative OD (%OD) was calculated in relation to the control incubated without blocking antibody. Each point indicates the average value of three independent experiments. Error bars indicate SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s HSD test for multiple comparison. An asterisk (*) indicates p < 0.01 at all antibody concentrations tested (0.1, 1, 2.5, 5, 7.5, and 10 μg/mL). n.s. , not significant. ( b , c ) Enhancement of cytokines production in equine PBMC cultures. Equine PBMCs ( n = 30) were obtained from healthy horse donors and stimulated with 0.1 μg/mL SEB in the presence of 10 μg/mL anti-PD-L1 antibody (Eqch6C11 or Eq6C11). Equine IgG was used as a control antibody. Culture supernatants were harvested on day 3, and concentration of ( b ) IFN-γ or ( c ) IL-2 was measured by ELISA. ( d , e ) Enhancement of cell proliferation in equine PBMC cultures. Equine PBMCs ( n = 12) collected from clinically healthy horses were cultured in the same condition as the cytokines production assay for 3 days. To evaluate cell proliferation, PBMCs were incubated with EdU during the last 2 h before the harvest. The lymphocyte population was gated by forward scatter and side scatter, and the incorporation of EdU in ( d ) CD4 + or ( e ) CD8 + cells was evaluated by a flow cytometer. ( b – e ) The gray bar represents the median value of each treatment group. Statistical analysis was performed using the Wilcoxon signed-rank test with Holm’s adjustment for multiple comparison. A single asterisk (*) or a double asterisk (**) indicate p < 0.05 or p < 0.01 respectively. n.s. , not significant.

Article Snippet: To evaluate cytokine production from cultured PBMCs, the culture supernatant was harvested on day 3, and the concentrations of interferon gamma (IFN-γ) and interleukin 2 (IL-2) were measured by enzyme-linked immunosorbent assay (ELISA) using the Equine IFN-γ ELISA Development Kit (HRP) (Mabtech, Nacka Strand, Sweden) and the Equine IL-2 DuoSet ELISA (R&D Systems, Minneapolis, MN, USA), respectively.

Techniques: Binding Assay, Inhibition, Blocking Assay, Activity Assay, Negative Control, Control, Incubation, Comparison, Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Flow Cytometry