Evaluation of a CD25-specific immunotoxin for prevention of graft-versus-host disease after unrelated marrow transplantation  Paul J. Martin, Ji Pei, Ted Gooley, Claudio Anasetti, Frederick R. Appelbaum, Joachim Deeg, John A. Hansen, Richard A. Nash, Effie W. Petersdorf, Rainer Storb, Victor Ghetie, John Schindler, Ellen S. Vitetta  Biology of Blood and Marrow Transplantation  Volume 10, Issue 8, Pages 552-560 (August 2004) DOI: 10.1016/j.bbmt.2004.04.002

Figure 1 Cyclosporine suppresses the expression of CD25 by responder cells in mixed lymphocyte culture. Responder cells were distinguished from stimulator cells with the use of an informative HLA marker, and dead cells were gated out by staining with 7-amino-actinomycin D. Panels A and B show forward light scatter and CD25 staining for cells cultured in the absence (A) or presence (B) of cyclosporine (400 ng/mL) on day 5. The vertical line indicates the distinction between blasts and nonactivated cells, and the horizontal lines indicate the distinctions among the CD25−, CD25-dull, and CD25-bright populations. The percentages of CD25-bright () and CD25-dull (▴) cells were analyzed on days 4 (C and E) and 5 (D and F) both in the whole population (C and D) and separately in blasts identified by high forward light scatter (E and F). Panels C and D also display the total percentage of CD25+ cells (□), whereas panels E and F display the percentage of blasts (□) in the cultures. Biology of Blood and Marrow Transplantation 2004 10, 552-560DOI: (10.1016/j.bbmt.2004.04.002)

Figure 2 Depletion of CD25+ cells on day 4 from a mixed lymphocyte culture in the presence of cyclosporine increased the secondary proliferative response. Cultures were maintained in the absence (A) or presence (B) of cyclosporine (400 ng/mL) and were depleted of CD25 cells (□) or were sham-depleted (). Secondary proliferative responses were tested in the absence of cyclosporine. Data indicate the mean ± SEM 3H-thymidine incorporation in triplicate cultures. Biology of Blood and Marrow Transplantation 2004 10, 552-560DOI: (10.1016/j.bbmt.2004.04.002)