Journal: PLoS ONE

Article Title: Fibrinogen-Like Protein 2/Fibroleukin Induces Long-Term Allograft Survival in a Rat Model through Regulatory B Cells

doi: 10.1371/journal.pone.0119686

Figure Lengend Snippet: (A) Cardiac graft recipients received intravenously 3x10 12 vector genomes/kg of AAV-FGL2 (▲ n = 8), or non coding AAV (▼ n = 5, 2 different experiments), and received a heterotopic transplant 30 days later. Graft survival was evaluated by palpation through the abdominal wall. Log-rank (Mantel-Cox) test *** p<0 . 001 for AAV-FGL2 vs. AAV null controls. (B) Left: Relative proportion of dividing CD4 + CD25 − T cells at day 6 in the presence of different concentrations of recombinant human FGL2-GST was evaluated by CFSE dilution by gating first on DAPI - live cells and then on TCR + CD4 + cells . The negative control was purified rat IgG at 10 μg/ml (n = 4, ** p <0.01). Right: Representative histogram of relative proportion of dividing CD4 + T cell in the presence of 10μg/ml FGL2-GST protein (black line) or IgG control (grey).

Article Snippet: Serum complement was inactivated by heating prior to incubation with donor splenocytes at a 1/8 dilution and revelation with either anti-rat IgG1 (MCA 194, Serotec), anti-rat IgG2a (MCA 278, Serotec), or anti-rat IgG2b (MCA195, Serotec) and anti-Ms Ig-488 as an isotype control (ref 11029, Serotec).

Techniques: Plasmid Preparation, Recombinant, Negative Control, Purification, Control

Journal: PLoS ONE

Article Title: Fibrinogen-Like Protein 2/Fibroleukin Induces Long-Term Allograft Survival in a Rat Model through Regulatory B Cells

doi: 10.1371/journal.pone.0119686

Figure Lengend Snippet: Sera were collected from naive rats, or at the moment of rejection from rats treated with AAV-control or AAV-FGL2 (rejecting at < 30 days or > 120 days after transplantation) or receiving adoptive transfers (> 120 days after transplantation). Levels of donor-specific IgG1, IgG2a, and IgG2b antibodies were evaluated by cytofluorimetry and normalized to serum from naive rats (MFI / MFI syngeneic). Two way Anova, Bonferroni post test p value * <0.5; ** <0.01; ***<0.001.

Article Snippet: Serum complement was inactivated by heating prior to incubation with donor splenocytes at a 1/8 dilution and revelation with either anti-rat IgG1 (MCA 194, Serotec), anti-rat IgG2a (MCA 278, Serotec), or anti-rat IgG2b (MCA195, Serotec) and anti-Ms Ig-488 as an isotype control (ref 11029, Serotec).

Techniques: Control, Transplantation Assay