Rabbit atg

Autologous Treg expanded from cryopreserved 2^nd round stock were labeled with 2 μM CFSE or VPD450. Approximately 2×10⁷/kg Treg were injected intravenously (i.v.) into normal untreated or IS monkeys (on day 0) that had received 2 consecutive intravenous (i.v.) infusions of rabbit ATG (Genzyme, Boston, MA) 2 days apart (day −20 and day −18) at doses of 10 and 5 mg/kg and Tacrolimus from day −20 to −16 (target whole blood trough levels between 10 and 15 ng/ml) followed by rapamycin (LC Laboratories) monotherapy from day −15 (target trough levels between 5 and 15 ng/ml) through the duration of the experiment. On day 66 post-infusion for one normal untreated monkey and d50 post-infusion for one IS monkey, PBMC (3ml), inguinal lymph node (LN) (2–3), mesenteric LN (2–3) and spleen cells were stained for CD3, CD4, CD25, CCR7, CD62L, and CXCR3. Data were analyzed with FlowJo software (Tree Star) and graphed with GraphPad Prism (Graph Pad Software, San Diego).

Rabbit ATG (Genzyme; Boston, MA) was given intravenously (i.v.); slowly over 4 hours on days -3 and -1 before transplant and on days 6 and 13 post-transplant at doses of 10, 5, 5 and 5 mg/kg, respectively. Methylprednisolone was given immediately before each ATG infusion at doses of 5, 2.5, 2.5 and 2.5 mg/kg, respectively. Anti-IL-6R mAb (Actemra; Genetech, CA) was administered i.v. over 1 hour at 10 mg/kg on days -1, 6, 13 and 20, and then once every 4 weeks. Tacrolimus was given by intramuscular (i.m.) injection from day -3 to 14 (target whole blood trough levels: 10–15 ng/ml), followed by rapamycin (i.m.; LC Laboratories, Woburn, MA) from days 14 to 54 (target whole blood trough levels: 10–15 ng/ml), after which rapamycin was weaned slowly and discontinued completely on day 84. The immunosuppressive protocol is summarized in Figure 1.

The Wayne State University institutional review board approved this study. We retrospectively reviewed charts of patients who underwent allo-SCT at our institution from January 2005 to December 2011. The eligibility criteria included development of signs or symptoms of acute or chronic GI GVHD at any time after transplantation. These included development of otherwise unexplained nausea, vomiting, anorexia, abdominal pain or cramps, diarrhea, failure to thrive, and weight loss [9 (link),10 (link)]. Grade of GVHD was determined clinically based on established criteria [9 (link),10 (link)]. Variables assessed for risk factors of the development of CMV viremia and CMV gastroenteritis were age, gender, race, number of transplantations, donor (related or unrelated), HLA mismatch, graft source (peripheral blood or bone marrow), recipient/donor CMV IgG serostatus, underlying diagnosis, disease status at the time of transplantation, conditioning regimen, GVHD prophylaxis regimen, antithymocyte globulin (rabbit-ATG, Genzyme) use, acute GVHD grade and peak CMV qPCR.