Presented at:
ASHI 31st Annual Meeting, Washington, DC, October 17-21, 2005.
Increased production of the regulatory T-cell transcription factor Fox P-3m-RNA and CD4+CD25+ cells in Campath-1H and donor hematopoetic stem cell treated kidney transplant recipients
Cirocco R, Careno M, Mathew J, Valone T, Esquenazi V, Miller J.
Division of Transplantation, University of Miami; Department of Microbiology and Immunology, University of Miami; Miami Veterans Affairs Medical Center. Miami, Florida.
Campath (C1H) is a potent immunosuppressant in kidney transplant recipients (KR). Donor Hematopoetic Stem Cells (DHSC) introduced after C1H treatment may increase the number of regulatory T-cells and facilitate engraftment. FOX P-3 (FP3) mRNA copy numbers per 5000 CD3+ cells from individuals in five different groups were measured:
- Normal laboratory volunteers and dialysis patients (found to be equivalent-'Normals') (N=21);
- C1H and DHSC treated KR (N=2) (Immune Tolerance Network Protocol);
- C1H treated KR (n=13);
- Thymoglobulin treated KR(Thy) (n=10);
- Zenopax treated KR (n=9).
CD3+ lymphocytes were isolated from whole blood, RNA extracted, C-DNA produced, analyzed for levels of FP3 m-RNA by real time PCR, and GAPDH standardized.
There were statistically significant higher levels of FP3 copies from the C1H + DHSC patients vs the C1H patients (p= .025). There were also higher levels in the C1H + DHSC patients or C1H patients when compared to 'Normals' (p=.009 and p= .002 respectively). C1H patients had significantly higher FP3 levels when compared to those of the Thy patients (p=.038). All other statistical comparisons were not significant. An increase (p=0.001) of T-regs or CD4+CD25+ cells was seen by flow cytometry in the C1H and Thy groups up to 240 days post transplant and more so in the C1H +DHSC patients.
In conclusion the use of C1H and (more so) of C1H + DHSC significantly increases the levels of Fox P-3 transcripts and CD4+ CD25+ cells in kidney transplant recipients.