Journal: PLoS ONE

Article Title: Induction of Foxp3-Expressing Regulatory T-Cells by Donor Blood Transfusion Is Required for Tolerance to Rat Liver Allografts

doi: 10.1371/journal.pone.0007840

Figure Lengend Snippet: (A) Representative flow cytometric analysis of splenocytes harvested from either naïve, DSTx4 alone or DSTx4+LTx rats 100days after LTx. Cells are gated on CD3 + CD4 + T-cells. Isotype-matched control antibodies were used to set all gates. (B) Percentages of Foxp3 + , CD25 + or CD45RC − T-cells within the CD4 + T-cell population in spleens from naive, DST treated or DSTx4+LTx rats. (C) Percentages of Foxp3 + CD25 − , Foxp3 + CD25 + or Foxp3 − CD25 + T-cells within the CD4 + T-cell population in spleens from naïve, DST treated or DSTx4+LTx rats. The results are expressed as mean±SE from at least three individual experiments for each group. * # ¶ P<0.05 compared with corresponding population in naïve group.

Article Snippet: PerCP conjugated mouse anti-rat CD8a (OX-8), FITC conjugated mouse anti-rat CD25 (OX-39), FITC conjugated mouse anti-rat CD45RC (OX-22), PE conjugated mouse anti-rat CD45RC (OX-22) and APC conjugated mouse anti-rat CD3 (1F4) antibodies were purchased from BD Bioscience (San Jose, CA).

Techniques:

Journal: PLoS ONE

Article Title: Induction of Foxp3-Expressing Regulatory T-Cells by Donor Blood Transfusion Is Required for Tolerance to Rat Liver Allografts

doi: 10.1371/journal.pone.0007840

Figure Lengend Snippet: Message levels of TGFβ, IL-10 and IFNγ in CD4 + Foxp3 + CD25 + T-cells obtained from spleens of long term survivors (DSTx4+LTx rats; survival>100 days) were quantified and compared to those expressed by CD4 + Foxp3 − CD25 + T-cells obtained from these same animals. Quantitative Real-time (RT)-PCR was used to quantify cytokine mRNA expression in each population. HRPT was used as an endogenous control to normalize each sample. Data represent mean±SD. * P<0.05 compared with Foxp3 − CD25 + T-cells. N = 5 for each group.

Article Snippet: PerCP conjugated mouse anti-rat CD8a (OX-8), FITC conjugated mouse anti-rat CD25 (OX-39), FITC conjugated mouse anti-rat CD45RC (OX-22), PE conjugated mouse anti-rat CD45RC (OX-22) and APC conjugated mouse anti-rat CD3 (1F4) antibodies were purchased from BD Bioscience (San Jose, CA).

Techniques: Quantitative RT-PCR, Expressing

Journal: PLoS ONE

Article Title: Induction of Foxp3-Expressing Regulatory T-Cells by Donor Blood Transfusion Is Required for Tolerance to Rat Liver Allografts

doi: 10.1371/journal.pone.0007840

Figure Lengend Snippet: (A) Adoptive transfer of whole splenocytes from DSTx4+LTx rats induces tolerance to liver allografts in naïve LEW recipients in a time-dependent manner. Splenocytes (250×10 6 cells) obtained from naïve (black diamonds, n = 7), DSTx4 (white diamonds,n = 9), DSTx4+LTx at 7 d post LTx (white circles, n = 4), DSTx4+LTx at 14 d post LTx (black circles, n = 5), DSTx4+LTx at 30 d post LTx (white squares, n = 4), DSTx4+LTx at 60 d post LTx (hash squares, n = 4) or DSTx4+LTx at 100 d post LTx (black squares, n = 9) were injected into lightly-irradiated (100rad) LEW rats one day before LTx. Control rats received 100 rad irradiation and received LTx but no cell transfer (black triangles, n = 14). *P<0.05 compared with no cell transfer. (B) Adoptive transfer of CD4 + but not CD8 + T-cells obtained from DST-primed LEW rats induces tolerance to liver allografts in naive LEW recipients. CD4 + (white squares, n = 6) or CD8 + (white diamonds, n = 6) T-cells (50×10 6 cells) obtained from spleens of naïve rats and CD4 + (black squares, n = 6) or CD8 + (black diamonds, n = 6) T-cells (50×10 6 cells) obtained from spleens of DSTx4 rats were injected into lightly-irradiated (100 rad) LEW rats one day before LTx. # P = 0.05 compared with naïve CD4 + (white squares). (C) Adoptive transfer of CD4 + CD45RC − , CD4 + CD25 + or CD4 + CD25 − T-cells induces tolerance to liver allografts in naïve LEW recipients. CD4 + T-cells (50×10 6 cells; (black squares, n = 7); CD4 + CD45RC − T-cells(20×10 6 cells; black diamonds, n = 5); CD4 + CD45RC + T-cells (20×10 6 cells; white diamonds,n = 5); CD4 + CD25 − T-cells (40×10 6 cells; black circles, n = 5); and CD4 + CD25 + T-cells (10×10 6 cells, white circles, n = 5) from DSTx4+LTx at 100days post LTx were transferred into lightly irradiated LEW rats one day prior to LTx. *P<0.05 compared with no cell transfer (black triangles).

Article Snippet: PerCP conjugated mouse anti-rat CD8a (OX-8), FITC conjugated mouse anti-rat CD25 (OX-39), FITC conjugated mouse anti-rat CD45RC (OX-22), PE conjugated mouse anti-rat CD45RC (OX-22) and APC conjugated mouse anti-rat CD3 (1F4) antibodies were purchased from BD Bioscience (San Jose, CA).

Techniques: Adoptive Transfer Assay, Injection, Irradiation