Journal: The Journal of Experimental Medicine

Article Title: Several Different Cell Surface Molecules Control Negative Selection of Medullary Thymocytes

doi:

Figure Lengend Snippet: For CD28, CD5, and CD43, costimulation for Fas-independent apoptosis of HSA hi CD4 + 8 − cells in vitro fails with strong TCR ligation. HSA hi CD4 + 8 − cells from normal B6 or B6 lpr/lpr mice were cultured overnight with either a low (0.1 μg/ml; left) or high (10 μg/ml; right) concentration of anti-TCR mAb plus/minus anti-CD5, anti-CD28, or anti-CD43 mAbs (20 μg/ml). The data show mean percent differences in (Δ) apoptosis for triplicate cultures.

Article Snippet: The following mAbs were purchased from PharMingen: anti–TCR-β (H57-597, hamster IgG), anti-CD2 (RM2-5, rat IgG), anti-CD5 (53-7.3, rat IgG), anti–LFA-1 (CD11) (M17/4, rat IgG), anti-CD27 (LG.3A10, hamster IgG), anti-CD28 (37.51, hamster IgG), anti-CD40L (CD154, MR1, hamster IgG), anti-CD43 (S7, rat IgG), anti-CD48 (HM48-1, hamster IgG), anti-CD49d (very late antigen [VLA]-4; R1-2, rat IgG), anti-CD81 (2F7, hamster IgG), anti–thymic shared antigen (TSA)-1 (MTS35, rat IgG), anti–CTL-associated antigen (CTLA)-4 (CD152; UC10-4F10-11, hamster IgG), anti-CD95 (Fas; Jo2, hamster IgG), and Cy-Chrome–conjugated anti-CD4 (H129.19, rat IgG).

Techniques: In Vitro, Ligation, Cell Culture, Concentration Assay

Journal: The Journal of Experimental Medicine

Article Title: Several Different Cell Surface Molecules Control Negative Selection of Medullary Thymocytes

doi:

Figure Lengend Snippet: Costimulation-dependent apoptosis induced by TCR/CD28, TCR/CD5, or TCR/CD43 ligation in vitro is abolished by IL-7. Purified HSA hi CD4 + 8 − thymocytes from B6 lpr/lpr mice were cultured with either a low or high concentration of anti-TCR mAb plus/minus mAbs specific for CD28, CD5, or CD43; where indicated, IL-7 was added to the cultures at 100 U/ml. The data show mean levels of difference in (Δ) apoptosis for triplicate cultures. Gray bar, anti-TCR (0.1 μg/ml); dotted bar, anti-TCR (0.1 μg/ml) plus IL-7; black bar, anti-TCR (10 μg/ml).

Article Snippet: The following mAbs were purchased from PharMingen: anti–TCR-β (H57-597, hamster IgG), anti-CD2 (RM2-5, rat IgG), anti-CD5 (53-7.3, rat IgG), anti–LFA-1 (CD11) (M17/4, rat IgG), anti-CD27 (LG.3A10, hamster IgG), anti-CD28 (37.51, hamster IgG), anti-CD40L (CD154, MR1, hamster IgG), anti-CD43 (S7, rat IgG), anti-CD48 (HM48-1, hamster IgG), anti-CD49d (very late antigen [VLA]-4; R1-2, rat IgG), anti-CD81 (2F7, hamster IgG), anti–thymic shared antigen (TSA)-1 (MTS35, rat IgG), anti–CTL-associated antigen (CTLA)-4 (CD152; UC10-4F10-11, hamster IgG), anti-CD95 (Fas; Jo2, hamster IgG), and Cy-Chrome–conjugated anti-CD4 (H129.19, rat IgG).

Techniques: Ligation, In Vitro, Purification, Cell Culture, Concentration Assay

Journal: PLoS ONE

Article Title: CD73-A2a adenosine receptor axis promotes innate B cell antibody responses to pneumococcal polysaccharide vaccination

doi: 10.1371/journal.pone.0191973

Figure Lengend Snippet: 8 to 12-week-old WT and CD73 KO mice were immunized i.p. with Pneumovax 23 and sera were analyzed by ELISA for anti-PPS3 specific Abs at day 0, 7 and 28 after vaccination. On day 28, mice were challenged by i.p. injection of S . pneumoniae (10 5 CFU). (A) Anti-PPS3 IgM and IgG3 OD (450–570) readings are shown (means ± standard errors are shown; *: p<0.05; **: p<0.01 by Student’s T test). (B) Combined results from 2 experiments pooling 20 WT and 19 CD73 KO mice showing IgM and IgG3 mean reciprocal titers at day 7 and 28 (means ± standard errors are shown; **: p<0.01; by One-way ANOVA; brackets are shown when significantly different from day 7 and 28 for a given group). (C) Survival of infected mice was monitored (*: p<0.05 by log-rank test; survival was compared to non-vaccinated mice, unless indicated by brackets; n.s: not significant).

Article Snippet: Fluorochrome conjugated antibodies to rat anti-mouse CD19 (1D3), CD5 (53–7.3), CD23 (B3B4), CD73 (TY/11.8), CD43 (S7), CD21/35 (7G6), rat IgG1,κ isotype control unconjugated mouse CD16/32 (24G2), and rat IgG1,κ anti-mouse IgA (C10-3) were obtained from BD Biosciences.

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Infection

Journal: PLoS ONE

Article Title: CD73-A2a adenosine receptor axis promotes innate B cell antibody responses to pneumococcal polysaccharide vaccination

doi: 10.1371/journal.pone.0191973

Figure Lengend Snippet: 8 to 12-week-old WT and A2a KO mice were immunized i.p. with Pneumovax 23 and sera were analyzed by ELISA for anti-PPS3 specific Abs on days 0, 7 and 28 after vaccination. On day 28, mice were challenged by i.p. injection of S . pneumoniae (10 5 CFU). (A) Anti-PPS3 IgM and IgG3 OD (450–570) readings are shown (means ± standard errors are shown; *: p<0.05; **: p<0.01 by Student’s T test). (B) Combined results from 2 experiments pooling 26 WT and 27 A2a KO mice showing IgM and IgG3 mean reciprocal titers at day 7 and 28 (means ± standard errors are shown; *: p<0.05; by One-way ANOVA; brackets are shown when significantly different from day 7 and 28 for a given group). (C) Survival of infected mice was monitored (*: p<0.05 by log-rank test; survival was compared to non-vaccinated mice, unless indicated by a bracket; n.s: not significant).

Article Snippet: Fluorochrome conjugated antibodies to rat anti-mouse CD19 (1D3), CD5 (53–7.3), CD23 (B3B4), CD73 (TY/11.8), CD43 (S7), CD21/35 (7G6), rat IgG1,κ isotype control unconjugated mouse CD16/32 (24G2), and rat IgG1,κ anti-mouse IgA (C10-3) were obtained from BD Biosciences.

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Infection

Journal: PLoS ONE

Article Title: CD73-A2a adenosine receptor axis promotes innate B cell antibody responses to pneumococcal polysaccharide vaccination

doi: 10.1371/journal.pone.0191973

Figure Lengend Snippet: Young (3-week-old) WT mice were immunized i.p. with 10 μg of Pneumovax 23 and were subsequently treated daily with CGS 21680 or vehicle only DMSO (control) for 4 weeks as described in methods. Sera were analyzed by ELISA for anti-PPS3 specific Abs at day 0, 7, 14 and 28 after vaccination. On day 28, mice were challenged by i.p. injection of S . pneumoniae (5x10 3 CFU). (A) Anti-PPS3 IgM and IgG3 OD (450–570) readings are shown (means ± standard errors are shown; *: p<0.05; **: p<0.01 by Student’s T test). (B) IgG3 mean reciprocal titers at day 7, 24 and 28 showing individual mice (means ± standard errors are shown; *: p<0.05; by One-way ANOVA). (C) Survival of infected mice (*: p≤0.05 by log-rank test; n.s.: p>0.05; survival was compared to non-vaccinated mice unless indicated by a bracket).

Article Snippet: Fluorochrome conjugated antibodies to rat anti-mouse CD19 (1D3), CD5 (53–7.3), CD23 (B3B4), CD73 (TY/11.8), CD43 (S7), CD21/35 (7G6), rat IgG1,κ isotype control unconjugated mouse CD16/32 (24G2), and rat IgG1,κ anti-mouse IgA (C10-3) were obtained from BD Biosciences.

Techniques: Enzyme-linked Immunosorbent Assay, Injection, Infection