Journal:

Article Title: Lipopolysaccharide from enterohemorrhagic Escherichia coli binds to platelets through TLR4 and CD62 and is detected on circulating platelets in patients with hemolytic uremic syndrome

doi: 10.1182/blood-2005-08-3219

Figure Lengend Snippet: LPS-induced platelet activation markers and binding of fibrinogen to human platelets detected by flow cytometry

Article Snippet: Detection of platelet activation was performed in resting platelets with hamster anti-mouse CD40L (10 μg/mL; BD Biosciences), followed by mouse anti-hamster-FITC (1:80; BD Biosciences).

Techniques: Activation Assay, Binding Assay, Flow Cytometry, Expressing

Journal:

Article Title: Lipopolysaccharide from enterohemorrhagic Escherichia coli binds to platelets through TLR4 and CD62 and is detected on circulating platelets in patients with hemolytic uremic syndrome

doi: 10.1182/blood-2005-08-3219

Figure Lengend Snippet: Expression CD40L on mouse platelets detected by flow cytometry

Article Snippet: Detection of platelet activation was performed in resting platelets with hamster anti-mouse CD40L (10 μg/mL; BD Biosciences), followed by mouse anti-hamster-FITC (1:80; BD Biosciences).

Techniques: Expressing, Flow Cytometry, Incubation

Journal: Immunity

Article Title: Low-Avidity CD4 + T Cell Responses to SARS-CoV-2 in Unexposed Individuals and Humans with Severe COVID-19

doi: 10.1016/j.immuni.2020.11.016

Figure Lengend Snippet:

Article Snippet: CD154-APC-Vio770 (clone: 5C8) , Miltenyi Biotec , Cat#130-114-130; RRID: AB_2751205.

Techniques: Functional Assay, Recombinant, Staining, Sequencing, Gene Expression, Software

Journal: International immunology

Article Title: Induction of Th1-biased cytokine production by alpha-carba-GalCer, a neoglycolipid ligand for NKT cells.

doi: 10.1093/intimm/dxq012

Figure Lengend Snippet: Fig. 3. Mechanisms of a-carba-GalCer-induced IFN-c production. (A and B) The levels of serum IL-12p70 and IFN-c after intravenous injection of a-GalCer (2 lg) (A) or a-carba-GalCer (2 lg) (B) were measured by ELISA or the cytometric bead array system at the indicated time points. Indicated antibodies were intraperitoneally injected 24 h before glycolipid injection. Data are means 6 SDs from three mice and repeated three times with similar results. (C and D) IL-12p70 production 6 h after intravenous injection of PBS, a-GalCer (2 lg) or a-carba-GalCer (2 lg) by spleen cells (C) and by splenic DC subsets (D). (C) Spleen cells were collected 1 h after glycolipid injection and cultured for 5 h in vitro. IL-12p70 production was analyzed by intracellular cytokine staining. (D) CD11c+ cells in (C) were further divided into CD8a CDCs, CD8a+ CDCs and B220+ PDCs, and IL-12p70 production was analyzed using intracellular cytokine staining. (E) CD40–CD40L interaction and IL-12 dependent production of IFN-c from NKTcells by a-carba-GalCer-pulsed BMDCs. Glycolipid-loaded or -unloaded BMDCs (2 3 105 cells per 200 ll) were co- cultured with NKT cells (7 3 104 cells per 200 ll) in the presence or absence of indicated mAbs (10 lg ml1). IL-12p70 and IFN-c in the supernatant at the indicated time point were measured by cytometric bead array. Results are representative of those from three independent experiments.

Article Snippet: For in vivo neutralizing experiments, goat polyclonal IgG specific for anti-mouse IL12 and anti-mouse CD40L was used and purchased from R&D systems (Minneapolis, MN, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Cell Culture, In Vitro, Staining

Journal: Inflammation and Regeneration

Article Title: Evaluation of immunosuppression protocols for MHC-matched allogeneic iPS cell-based transplantation using a mouse skin transplantation model

doi: 10.1186/s41232-021-00190-7

Figure Lengend Snippet: Blockade of leukocyte costimulatory molecules permits long-term engraftment of mouse skin grafts from MHC-matched allogeneic donors. ( A ) CB-based immunoregulation protocol. Immunosuppression was performed by using a protocol of administrating a combination of anti-CD40L mAb and CTLA4-Ig daily (CB) with or without rapamycin on every 3 days from POD 1 (CB + rapa). ( B ) Skin graft survival under immunosuppression with CB ( n = 9, C3129F1; n = 9, BALB/c; n = 9, C57BL/6; n = 9, CBA/N). ( C ) Skin graft survival under immunosuppression with CB + rapa ( n = 9, C3129F1; n = 8, BALB/c; n = 9, C57BL/6; n = 9, CBA/N). ( D ) Macroscopic observation (upper panels) and hematoxylin and eosin stained section (lower panels) of autologous (C3129F1), CBA/N, and C57BL/6 skin grafts on day 100. Scale bars: 100 μm. ( E ) Immunohistochemical staining for CD3 of C57BL/6 skin grafts harvested from CB or CB + rapa treated groups on 100 days post-transplantation. Upper and lower panels represent CD3 specific staining and isotype control stained section, respectively. Scale bars: 100 μm. ( F ) Recipient T cell response in MHC-matched but minor antigen-mismatched skin transplantation. The T cell proliferation rates in each treatment group were normalized to that of C3129F1 mice stimulated with autologous irradiated splenocytes. Error bars indicate standard error of technical triplicates. Similar results were obtained in two independent experiments. * p < 0.05, ** p < 0.01 (Tukey’s HSD test). ( G ) De novo anti-donor antibody production in the recipients. Error bars indicate standard error of biological replicates ( n = 8, naive; n = 4, non-treatment; n = 9, CB; n = 9, CB + rapa). tx, transplantation; mAb, monoclonal antibody; CB, co-stimulatory molecule blocking; rapa, rapamycin; POD, post-operative day

Article Snippet: For co-stimulatory molecule blockade (CB) therapy, anti-CD40 ligand (CD40L) (MR-1 for mouse and #BE0292 for human, BioXcell) and CTLA4Ig (Orencia®, Bristol Myers Squibb for mouse and #BE0099, BioXcell for human) were administered at a dose of 500 μg for anti-CD40L, 400 μg for CTLA4Ig on days 0, 2, 4, and 6 after transplantation.

Techniques: Staining, Immunohistochemical staining, Transplantation Assay, Control, Irradiation, Blocking Assay