Relationship between abnormal p53 protein and failure to express p21 protein in human breast carcinomas. the estimated 5-year survival rate was 51%. In a multivariate MLN 0905 model, a CD4 count of 100/mm3 and a below normal serum immunoglobulin G level at study entry were associated with a short survival duration ( .0001). CONCLUSIONS These results confirm the potential cure of relapsed/refractory CLL with NST and provide the first evidence that immunoglobulin G and CD4 levels are predictive of overall survival after NST in CLL and that human leukocyte antigen alleles predict response to immunomanipulation. mutation in CLL.18 We retrospectively performed immunohistochemical studies of 200 nuclei using routinely fixed and processed bone marrow trephine biopsy samples and heat-induced epitope retrieval, as described previously.19 We chose bone marrow samples that were collected within 3 months pre-NST. All cases were scored independently by at MLN 0905 least 2 researchers. Neoplasms (PAX 5+) with p53 staining in 20% of tumor cells were considered positive. Neoplasms with p21 staining in 5% of tumor cells were considered negative, as previously suggested.18 Preparative Regimens, Transplantation, and Post-Transplantation Immunomanipulation Details of the preparative regimens, supportive care, and infection and GVHD prophylaxis and early results have been previously published.1,7,12 The predominant preparative regimen used was 30 mg/m2 of fludarabine, 750 mg/m2 of cyclophosphamide given intravenously on Days ?5 to ?3 before transplantation, and 375 mg/m2 of rituximab on Day ?13 and 1000 mg/m2 on Days ?6, + 1, and +8 (n = 78). Eight patients received other preparative regimens (fludarabine, melphalan, and rituximab [n = 6] and rituximab, carmustine, etoposide, cytarabine, and melphalan [n = 2]). Forty-three (50%) MLN 0905 patients received transplants from histocompatible siblings, and 43 (50%) who had no matched sibling donors received transplants from unrelated donors. GVHD prophylaxis consisted of tacrolimus and methotrexate (5 mg/m2 on Days +1, +3, and +6, with an additional dose on Day +11 for nonsibling transplants) as previously described.1,7,12 Post-transplantation immunomanipulation (with-drawal of immunosuppression, infusion rituximab, and step-wise DLI) was performed in patients with MLN 0905 progressive or residual disease using previously established methods.7,12 Rituximab was given at a dose of 375 mg/m2 intravenously, followed by 3 weekly doses of 1000 mg/m2. A DLI of 1 1 106 CD3-positive T cells/kg was administered after the first 2 doses of rituximab if no GVHD occurred. An escalated DLI dose was given at 6-week intervals if there was persistent active disease and no MLN 0905 GVHD. DLIs were not routinely administered in patients with stable mixed chimerism if they remained in remission. Acute and chronic GVHD was graded according to consensus criteria.20 GVHD after DLI was graded according to National Institutes of Health consensus criteria.21 Immune Assays During Immunomanipulation To determine the immune parameters related to response, we monitored natural killer (NK) activity in 13 patients who had undergone immunomanipulation. This prospective analysis was undertaken on heparinized blood samples collected at 5 time points: prestudy entry, before each of the first 2 doses of rituximab, before DLI, and then after the last dose of rituximab given with immunomanipulation. DLIs were provided between the second and third doses of rituximab. The blood samples were obtained according to an institutional review board-approved protocol at The University of Texas MD Anderson Cancer Center. Statistical Analysis Disease and transplantation characteristics were compared by using Fisher exact test for categorical variables and the Mann-Whitney test for continuous variables. Actuarial estimates of overall survival (OS) rates were calculated using the Kaplan-Meier method.22 The incidences of disease progression and GVHD were estimated using the cumulative incidence method, 23 considering death with disease and death without GVHD as competing risks. The current progression-free survival (PFS) rate, accounting for salvage therapy after DLI, was estimated using a linear combination of Kaplan-Meier estimates, as described by Klein et al.24 Risk factors for disease progression and death were evaluated using Cox proportional hazards. Logistic regression was utilized to assess predictors of response to immunomanipulation. Precise logistic regression was found in analyses concerning a small amount of occasions. For proportional risks and logistic regression analyses, univariate versions primarily had been match, and everything factors which were significant had been fit jointly in multivariate analyses statistically. Model selection was performed by backward stepwise regression.25 The utmost statistically E2F1 significant value was .05. Outcomes Research Group Eighty-six individuals (70 males and 16 ladies).