Human granulocyte colony-stimulating factor (G-CSF) is a regulatory glycoprotein that stimulates the production of neutrophilic granulocytes from committed hematopoietic progenitor cells both in vitro and in vivo. In this report, we show that biosynthetic (recombinant) human G-CSF enhances colony formation by normal human bone marrow and the human myeloid leukemic cell lines, HL-60 and KG-1, as well as nonhematopoietic small cell lung cancer lines, H128 and H69. G-CSF also modulates multiple differentiated functions of human neutrophils, including enhanced oxidative metabolism in response to f- Met-Leu-Phe (f-MLP), increased antibody-dependent cell-mediated cytotoxicity (ADCC), and augmented arachidonic acid release in response to ionophore and chemotactic agents. These effects are all maximal at a concentration of 100 to 500 pmol/L. Using 125I-labeled recombinant human G-CSF, high affinity binding sites were identified on human neutrophils, the myeloid leukemia cell lines KG-1 and HL-60, and the small cell carcinoma cell lines, H128 and H69. G-CSF receptor numbers ranged between 138 and 285 sites per cell with a kd of 77 to 140 pmol/L, consistent with the concentrations of G-CSF that elicit biologic responses in vitro. Decreased specific binding of 125l-G-CSF by human neutrophils was consistently observed in the presence of excess unlabeled human granulocyte-macrophage colony-stimulating factor (GM-CSF), suggesting competition or down modulation by GM-CSF of the G- CSF receptor.

One hundred and thirty-six patients autografted for relapsed or refractory non-Hodgkin's lymphoma (NHL) were evaluated to assess long-term event-free survival and to identify important prognostic factors. High-dose therapy consisted primarily of carmustine (BCNU), etoposide, cytarabine, and cyclophosphamide (BEAC) followed by unpurged autologous stem cell rescue. The 5-year Kaplan-Meier event-free survival (EFS) for the entire cohort was 34% (95% confidence interval: 24-44%) with a median follow-up of approximately 3 years (range 0-7.5 years). For patients entering with minimal disease (defined as all areas < or = 2 cm), the 5-year EFS was 40 vs 26% for those entering with bulky disease (P = 0.0004). In the multivariate analysis, minimal disease on entry and administration of involved-field XRT post-transplant were significantly associated with improved EFS; the latter association was observed mainly in the cohort of patients with bulky disease. The overall 100-day treatment-related mortality rate was 4.4% (3% for the last 71 patients). New strategies are needed to reduce the high rate of relapse (50-60%) following auto-transplantation for relapsed or refractory NHL.

Seventy consecutive patients with refractory or relapsed Hodgkin's disease who received high-dose chemotherapy followed by autologous stem cell rescue were analyzed to identify clinically relevant predictors of long-term event-free survival. High-dose therapy consisted primarily of carmustine (BCNU), etoposide, cytarabine and cyclophosphamide (BEAC). The 5-year Kaplan-Meier event-free survival (EFS) for the entire cohort was 32% (95% confidence interval; 18-45%) with a median follow-up of 3.6 years (range 7 months-7.6 years). The most significant predictor of improved survival was the presence of minimal disease (defined as all areas < or =2 cm) at the time of transplant: the 5 years EFS was 46 vs 10% for patients with bulky disease (P = 0.0002). Other independent predictors identified by step-wise regression analysis included the presence of non-refractory disease and the administration of post-transplant involved-field radiotherapy (XRT). Treatment-related mortality occurred in 13 of 70 patients: nine patients (13%) died within the first 100 days, mainly from cardiopulmonary toxicity. However, only one of 24 patients (4%) transplanted during the last 4.5 years died from early treatment-related complications. While high-dose therapy followed by autotransplantation led to long-term EFS of 50% for patients with favorable prognostic factors, a substantial proportion of patients relapsed, indicating that new therapeutic strategies are needed.

Summary:We report two cases of severe alloimmune hemolysis after hematopoietic stem cell (HSC) transplant resulting from an anti-Jk a . The time course of hemolysis and Jk phenotypes of the donor and recipient in the cases reported suggest that the antibody was produced by donor-derived passenger lymphocytes. Retrospective analysis of the blood bank records of all allogeneic HSC transplant patients at Barnes-Jewish Hospital from 1994 to 1999 suggests that the incidence of alloimmune hemolysis due to incompatibilities involving non-ABO or RhD red cell antigens is very low, approximately 1%. In one patient, the duration of hemolysis was shortened significantly by performing red cell exchange at the first sign of intravascular hemolysis. Bone Marrow Transplantation (2001) 27, 1305-1310. Keywords: allotransplantation; hemolysis; alloantibodies; stem cells Delayed immune hemolysis is an uncommon, but serious complication of HSC transplantation. Pretransplant clinical consideration is usually given only to ABO and Rh compatibility between the donor and recipient. [1][2][3][4] The occurrence of immune hemolyis in HSC transplant recipients who receive ABO or RhD-mismatched marrow has been estimated to be approximately 10% to 15%. 3 For example, in cases of major ABO mismatch, care is taken to red cell deplete the stem cell product prior to transplant. In the absence of pre-existing alloantibodies to non-ABO antigens in the donor or recipient, no further consideration is given prior to transplant to a potential non-ABO/RhD antigen mismatch.There are only a handful of reports in the literature reporting occurrences of non-ABO alloimmunization after allogeneic transplantation and only a few of these cases occurred following transplantation of HSCs derived either from bone marrow (BM) or peripheral blood mononuclear cells (PBMC) isolated by apheresis. 5-9 These reports suggest that, in contrast to patients with posttransplant hemolysis due to mismatches in the ABO system in whom hemolysis is usually moderate, non-ABO alloimmune hemolysis in the post HSC transplant setting was associated with episodes of severe hemolytic anemia presenting difficult clinical management issues. 8 No estimates of the frequency of non-ABO/RhD red cell alloimmunization in the post-transplant setting are available in the published literature.We present two case reports of severe immune hemolytic anemia after allogeneic HSC transplant resulting from donor-derived anti-Jk a . We also retrospectively reviewed the blood bank records of all patients at Barnes-Jewish Hospital who underwent allogeneic HSC transplantation (either BM or PBMC) between January 1994 and December 1999 to determine the incidence of non-ABO alloantibodymediated hemolysis in this population. Case 1The first patient was a 37-year-old white male with Philadelphia chromosome positive chronic myelogenous leukemia who presented with a WBC of 31 000. He was managed with hydroxyurea with adequate control for 2 years. Upon progression, a decision was made to proceed with a 6/6 HLA-matche...