Journal: Signal Transduction and Targeted Therapy

Article Title: Eosinophils promote CD8 + T cell memory generation to potentiate anti-bacterial immunity

doi: 10.1038/s41392-024-01752-0

Figure Lengend Snippet: IL-4 inhibits the JNK/caspase-3-mediated CD8 + T cell apoptosis in response to L.m . infection. a – c Representative FACS image and statistical analysis of TLR4-MD2 + ( a ), Annexin-V + ( b ) and PD-1 + ( c ) CD8 + T cells with or without LLO treatment for 4–6 h. d – f Representative FACS image and statistical analysis of Annexin-V + ( d ), PD-1 + ( e ) and cytoplasmic p-JNK + ( f ) of CD8 + T cells with or without JNK inhibitor IQ-1S treatment after L.m . infection. g – i Representative FACS image and statistical analysis of Annexin-V + ( g ), PD-1 + ( h ) and cytoplasmic p-JNK + ( i ) of CD8 + T cells with or without exogenous IL-4 treatment after L.m . infection. j Representative FACS image and quantification of MFI of IL-4R expression in CD8 + T cells with or without treatment of exogenous IL-4. k , l Proportions of Annexin-V + and PD-1 + CD8 + T cells with or without Z-VAD-FMK ( k ) and Z-DEVD-FMK ( l ) treatment after L.m . infection as measured by flow cytometry. m Proportions of cleaved Caspase-3 + CD8 + T cells with or without IL-4 treatment after L.m . infection as measured by flow cytometry. n CFSE analysis of proliferative CD8 + T cells after anti-CD3/CD28 stimulation with IL-4 treatment or not. Data are mean ± SD of one representative experiment. Similar results were seen in two or three independent experiments. Unpaired Student’s t tests. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Cell Activation Cocktail (with Brefeldin A), Fixation Buffer, APC-Cy7 anti-mouse CD45 (30-F11), PE-Cy7 anti-mouse CD45.1 (A20), PE-Cy7/ Percp-cy5.5 anti-mouse CD4 (RM4-5), APC anti-mouse IFN-γ (XMG1.2), APC anti-mouse PD-1 (29F.1A12), APC/BV605 anti-mouse KLRG1 (2F1), APC anti-mouse CX3CR1 (SA011F11), APC/AF488 anti-mouse CD11b (M1/70), Percp-cy5.5/BV421 anti-mouse CD8a (53-6.7), BV421 anti-mouse Siglec-F (S17007L), purified anti-mouse IL-4 antibody (11B11), PE-Cy7 anti-mouse TLR4-MD2 (MTS510), PE anti-mouse IL-4 (11B11), rmCXCL3, rmCXCL5 were purchased from Biolegend.

Techniques: Infection, Expressing, Flow Cytometry

Journal: PLoS ONE

Article Title: Augmented TLR2 Expression on Monocytes in both Human Kawasaki Disease and a Mouse Model of Coronary Arteritis

doi: 10.1371/journal.pone.0038635

Figure Lengend Snippet: (A) Representative histograms showed surface expression of TLR2 and TLR4 by mean fluorescence intensity (MFI) on peripheral monocytes, neutrophils, and lymphocytes in KD patients before IVIG treatment and control patients. (B) The representative histogram displayed surface expression of TLR2 by MFI on peripheral CD14 + monocytes in human KD patients before and after IVIG treatment as well as those in control patients. (C) TLR2 expression over CD14 + monocytes are expressed as mean±SEM by MFI. n = 6 in controls; n = 12 Pre-IVIG; n = 8 Post-IVIG. ** P <0.01.

Article Snippet: Each staining reaction was incubated for 30 minutes with the following primary antibodies and their corresponding isotypes (all from eBioscience, San Diego, CA) at 4°C as follows: fluorescein isothiocyanate (FITC)-conjugated anti-human CD14, Phycoerythrin (PE)-conjugated anti-human TLR2, PE-conjugated anti-human TLR4, allophycocyanin-labeled anti-mouse CD14, biotin-conjugated anti-mouse TLR2 (subsequently being labeled with streptavidin-peridinin-chlorophyll protein complex), and PE-conjugated anti-mouse TLR4/MD2.

Techniques: Expressing, Fluorescence

Journal: PLoS ONE

Article Title: Augmented TLR2 Expression on Monocytes in both Human Kawasaki Disease and a Mouse Model of Coronary Arteritis

doi: 10.1371/journal.pone.0038635

Figure Lengend Snippet: (A) Representative histograms showed surface expression of TLR2 and TLR4/MD2 by MFI on monocytes, neutrophils, and lymphocytes in the LCWE-treated mice and PBS-treated control mice on post-injection day 7. The numbers labeled over the right upper quadrants indicated the representative percentage of TLR2 + or TLR4/MD2 + cells, corresponding to each cell subpopulation. (B) The surface expression of TLR2 on circulating CD14 + monocytes were analyzed by relative TLR2 MFI (rMFI = TLR2 MFI/isotype MFI) at the indicated time points post-injection. (C) Circulating CD14 + monocytes were significantly increased in LCWE-treated mice on days 3, 7, and 14 post injections. Values are expressed as mean±SME, n = 7 mice/group per time point; * P <0.05; ** P <0.01 versus PBS controls.

Article Snippet: Each staining reaction was incubated for 30 minutes with the following primary antibodies and their corresponding isotypes (all from eBioscience, San Diego, CA) at 4°C as follows: fluorescein isothiocyanate (FITC)-conjugated anti-human CD14, Phycoerythrin (PE)-conjugated anti-human TLR2, PE-conjugated anti-human TLR4, allophycocyanin-labeled anti-mouse CD14, biotin-conjugated anti-mouse TLR2 (subsequently being labeled with streptavidin-peridinin-chlorophyll protein complex), and PE-conjugated anti-mouse TLR4/MD2.

Techniques: Expressing, Injection, Labeling