Journal: Immunity

Article Title: Secondary influenza challenge triggers resident memory B cell migration and rapid relocation to boost antibody secretion at infected sites

doi: 10.1016/j.immuni.2022.03.003

Figure Lengend Snippet: Alveolar plasma cells are derived from memory B cells (A) Experimental outline for (A–C). (B) TPLSM images of CD19 −/− mice transferred with BAT B cells as described in (A) before and after rechallenge. PCs are highlighted using Imaris-created spots (yellow). (C) PCs L values at r = 200. (D) Alveolar PC density in infected or uninfected sites. Right, data represented as the fold-change difference. (E) Left, experimental outline. Right, PCs L values in untreated resting memory mice or rechallenged mice treated with anti-CD40L. Statistical analysis were made using an unpaired t test (C), a paired t test (D), and a Mann-Whitney U test (E). Error bars represent SD. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. See also Figure S4 .

Article Snippet: InVivoMAb anti-mouse CD40L, Clone MR-1 , BioXCell , Cat# BE0017-1; RRID: AB_1107601.

Techniques: Derivative Assay, Infection, MANN-WHITNEY

Journal: Immunity

Article Title: Secondary influenza challenge triggers resident memory B cell migration and rapid relocation to boost antibody secretion at infected sites

doi: 10.1016/j.immuni.2022.03.003

Figure Lengend Snippet:

Article Snippet: InVivoMAb anti-mouse CD40L, Clone MR-1 , BioXCell , Cat# BE0017-1; RRID: AB_1107601.

Techniques: Purification, Clone Assay, Generated, Recombinant, Expressing, Software, Imaging, Light Microscopy

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: JCI Insight

Article Title: Allograft dendritic cell p40 homodimers activate donor-reactive memory CD8 + T cells

doi: 10.1172/jci.insight.96940

Figure Lengend Snippet: Groups of C57BL/6 (n = 4/group) mice received A/J cardiac allografts subjected to 8 hours of CIS. On days 0 and 1 after transplant, all recipients were injected with 100 μg BrdU i.p. and either 250 μg control rat IgG or (A) 250 μg CTLA-4Ig or (B) 250 μg anti-CD154 mAb. After 48 hours, allografts were harvested and digested, and aliquots of single cell suspensions were stained with antibody and analyzed by flow cytometry, with examples of gating as shown for each allograft sample to assess and quantitate the BrdU incorporation of infiltrating memory CD4+ and CD8+ T cells.***P < 0.001, as determined by the Mann-Whitney nonparametric test. (C) Groups of C57BL/6 recipient mice (n = 5–6/group) received A/J cardiac allografts subjected to 8 hours of CIS and, on days 0 and 1 after transplant, were treated with 250 μg control rat IgG, 250 μg CTLA-4Ig, or 250 μg anti-CD154 mAb i.p. Allograft survival was monitored by daily abdominal palpation, and rejection was confirmed by laparotomy. **P < 0.01 versus 8 hours of CIS + IgG or + CTLA-4Ig, as determined using the Log-rank/Mantel-Cox test.

Article Snippet: Anti-CD154 mAb (MR-1, Bio X Cell) was administered at 0.25 mg i.p. daily on days 0 and +1.

Techniques: Injection, Staining, Flow Cytometry, BrdU Incorporation Assay, MANN-WHITNEY

Journal: Vaccine

Article Title: A multi-trimeric fusion of CD40L and gp100 tumor antigen activates dendritic cells and enhances survival in a B16-F10 melanoma DNA vaccine model

doi: 10.1016/j.vaccine.2015.07.081

Figure Lengend Snippet: (a) Model of SPD-gp100-CD40L. Amino acids 25 to 596 (sequence KVPRNQD to EAGLGQV) of human gp100 were inserted between amino acids 105 and 106 of murine SPD within the SPD-CD40L fusion construct. (b) Cartoon of expected SPD-gp100-CD40L 4-trimer structure showing trimers of CD40L (gray circles) and trimers of gp100 (black circles) extending from the SPD collagen-like domain. (c) Western blot analysis. 293T cells were transfected with DNA plasmid encoding gp100 or SPD-gp100-CD40L fusion protein. After 48-hour culture, supernatant was collected and run on an SDS-PAGE gel in the presence of reducing agent. Western blot was performed using a polyclonal antibody to gp100.

Article Snippet: Following electrophoresis, protein was blotted onto PVDF membrane (Pierce) by electrophoretic transfer, blocked, and probed with goat anti-mouse CD40L antibody (R&D Systems).

Techniques: Sequencing, Construct, Western Blot, Transfection, Plasmid Preparation, SDS Page

Journal: Vaccine

Article Title: A multi-trimeric fusion of CD40L and gp100 tumor antigen activates dendritic cells and enhances survival in a B16-F10 melanoma DNA vaccine model

doi: 10.1016/j.vaccine.2015.07.081

Figure Lengend Snippet: (a) Representative SEC elution profile of SPD-gp100-CD40L as monitored by the scattering intensity, expressed in terms of the excess Rayleigh ratio (R(θ)), at three scattering angles of 42° (tall gray line), 90° (black line) and 138° (short gray line) as a function of elution volume (V). Note that SPD-gp100-CD40L apparently elutes as a major species (megamer) and as a minor species (polymer, evident as a shoulder of the major elution peak). (b) Representative partial Zimm plot obtained from analytical SLS measurements at a specific protein concentration for the SPD-gp100-CD40L complex. The line through the data points—collected at three scattering angles of 42°, 90° and 138°—represents a linear fit. (c) Representative autocorrelation function plot obtained from analytical DLS measurements at a specific protein concentration for the SPD-gp100-CD40L complex. The line through the data points—collected at a scattering angle of 90°—represents non-linear least squares fit.

Article Snippet: Following electrophoresis, protein was blotted onto PVDF membrane (Pierce) by electrophoretic transfer, blocked, and probed with goat anti-mouse CD40L antibody (R&D Systems).

Techniques: Polymer, Protein Concentration

Journal: Vaccine

Article Title: A multi-trimeric fusion of CD40L and gp100 tumor antigen activates dendritic cells and enhances survival in a B16-F10 melanoma DNA vaccine model

doi: 10.1016/j.vaccine.2015.07.081

Figure Lengend Snippet: (a) In vitro activity of SPD-CD40L and SPD-gp100-CD40L was determined using a cell-based CD40 NF-κB enzymatic reporter system. An equivalent amount of 293T supernatant from pcDNA3.1, pSPD-CD40L or pSPD-gp100-CD40L transfected cells was incubated with 293-CD40-SEAP NF-κB reporter cells at various dilutions. (b) In vitro activity of supernatant from SPD-gp100-CD40L transfected 293T cells was evaluated on mouse bone marrow derived mouse DC and compared to supernatant from 293T cells transfected with empty vector pcDNA3.1. Error bars represent mean + SEM (n=3). * p<0.05, ** p<0.01 by Student’s t test compared to pcDNA3.1 supernatant.

Article Snippet: Following electrophoresis, protein was blotted onto PVDF membrane (Pierce) by electrophoretic transfer, blocked, and probed with goat anti-mouse CD40L antibody (R&D Systems).

Techniques: In Vitro, Activity Assay, Transfection, Incubation, Derivative Assay, Plasmid Preparation

Journal: Vaccine

Article Title: A multi-trimeric fusion of CD40L and gp100 tumor antigen activates dendritic cells and enhances survival in a B16-F10 melanoma DNA vaccine model

doi: 10.1016/j.vaccine.2015.07.081

Figure Lengend Snippet: (a) Immunization schedule for B16-F10 tumor challenge and DNA/GVAX therapeutic vaccination, as indicated by arrows. B16-F10 cells (50,000) were injected i.d. into the left flank of mice on day 0. Mice were then immunized by i.m. injection of PBS or pSPD-gp100-CD40L on day 3, 10, and 17. GVAX, B16-F10 tumor cells expressing GM-CSF, were irradiated at 5,000 rad and 1 × 106 cells injected subcutaneously on day 3, 6, and 9. (b) Tumor growth analysis. Each point represents the mean tumor volume per group (n=5). (c) Survival analysis was based on the date of death or when tumor size reached >1500 cm2.

Article Snippet: Following electrophoresis, protein was blotted onto PVDF membrane (Pierce) by electrophoretic transfer, blocked, and probed with goat anti-mouse CD40L antibody (R&D Systems).

Techniques: Injection, Expressing, Irradiation

Journal: Vaccine

Article Title: A multi-trimeric fusion of CD40L and gp100 tumor antigen activates dendritic cells and enhances survival in a B16-F10 melanoma DNA vaccine model

doi: 10.1016/j.vaccine.2015.07.081

Figure Lengend Snippet: (a) Immunization schedule for B16-F10 tumor challenge and DNA or GVAX vaccination, as indicated by arrows. B16-F10 cells (50,000) were injected i.d. into the left flank of the mice on day 0. Mice were immunized i.m. with PBS, pSPD-gp100-CD40L + pIL-12, pSPD-gp100-CD40L + pGM-CSF, or pSPD-gp100-CD40L + pIL-12 + pGM-CSF on day 3, 10, and 17. For GVAX therapy, B16-F10 tumor cells expressing GM-CSF (GVAX), were irradiated at 5,000 rad and 1 × 106 cells were injected subcutaneously on day 3, 6, and 9. (b) Tumor growth analysis. Each point represents the mean tumor volume of animals in each group (n=5). (c) Survival analysis of mice. (d) Tumor growth kinetics of individual mice from each treatment arm.

Article Snippet: Following electrophoresis, protein was blotted onto PVDF membrane (Pierce) by electrophoretic transfer, blocked, and probed with goat anti-mouse CD40L antibody (R&D Systems).

Techniques: Injection, Expressing, Irradiation

Journal: Vaccine

Article Title: A multi-trimeric fusion of CD40L and gp100 tumor antigen activates dendritic cells and enhances survival in a B16-F10 melanoma DNA vaccine model

doi: 10.1016/j.vaccine.2015.07.081

Figure Lengend Snippet: (a) Immunization schedule for B16-F10 tumor challenge and DNA vaccination, as indicated by arrows. B16-F10 cells (50,000) were injected into the left flank of the mice on day 0. Mice were then immunized i.m. with PBS, pgp100, pgp100 + pIL-12, pgp100 + pGM-CSF, pgp100 + pIL-12 + pGM-CSF, pgp100-IRES-SPD-CD40L + pIL-12 + pGM-CSF, or pSPD-gp100-CD40L + pIL-12 + pGM-CSF on day 3, 10, and 17. (b) Tumor growth analysis. Each point represents the mean tumor volume of animals in each group (n=5). (c) Survival analysis of mice.

Article Snippet: Following electrophoresis, protein was blotted onto PVDF membrane (Pierce) by electrophoretic transfer, blocked, and probed with goat anti-mouse CD40L antibody (R&D Systems).

Techniques: Injection

Journal: Cell reports

Article Title: In Vivo Chimeric Alzheimer’s Disease Modeling of Apolipoprotein E4 Toxicity in Human Neurons

doi: 10.1016/j.celrep.2020.107962

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Immunosuppressants were comprised of a cocktail of anti-mouse CD40L (CD154) (BioXCell), anti-mouse CTLA-4 (CD152) (BioXCell), and anti-mouse LFA-1a (CD11a) (BioXcell), and all were used at a concentration of 20 mg/kg.

Techniques: Recombinant, Blocking Assay, Plasmid Preparation, Software