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Introduction Pediatric inspired regimens can achieve good outcomes in adolescents and young adults (AYAs) with acute lymphoblastic leukemia (ALL) by delivering intense non-myelosuppressive chemotherapy and are considered standard. Experience with the implementation of these regimens outside of academic centers in high-income countries is limited. Furthermore, Mexican patients (with "hispanic ethnicity") have increased risk for relapse and treatment related complications. Objectives Primary objective: to determine 2-year overall survival (OS) and event-free survival (EFS). Secondary objectives: to determine the impacts of treatment abandonment and measurable residual disease (MRD) on outcomes, and to compare treatment costs with a widely used standard regimen in the United States. Patients and Methods Consecutive patients 16-45 years diagnosed with Ph-negative B-cell acute lymphoblastic leukemia after 2016 were included. The salient features of our modified-BFM regimen include the use mitoxantrone, E. coli L-asparaginase, without systemic cytarabine or high dose methotrexate designed to be delivered entirely in an outpatient basis for maximum affordability (Table 1), given that we treat an uninsured population that must cover their own treatment costs out of pocket. Genetic risk assessment was limited to BCR/ABL. Thereafter, relapse risk assessment was based exclusively on "next generation" flow cytometry measurable residual disease (MRD) after consolidation, with a planned allogeneic transplant for MRD-positive patients (≥0.001%). Treatment abandonment was defined as a missed ≥14-day period during intensive treatment or ≥1 month during maintenance. Statistical analysis was performed as intent-to-treat. Lastly, drugs included in our protocol were compared to those of CALGB 10403 with current local pricing in USD. Results Ninety-one AYAs have been treated, 47 women and 44 men, with a median age of 21 years (range, 15-45), mostly with a good functional capacity and no comorbidities (ECOG≤2: 92.1%; HCT-Ci 0-1: 97.8%); 43.8% of patients had ≥30x10 9/L white blood cells at diagnosis and 31.7% had grade ≥1 obesity. Notable grade ≥3 adverse events during induction were infections/neutropenic fever (35.6%),hepatotoxicity (11%) and thrombosis/bleeding (8.1%) with 44.3% eventually requiring hospitalization. Induction related mortality was 11%. Only n=3 were refractory to induction and the remainder assessed achieved complete remission (n=63; 95.5%) with a median follow-up of 15 months (range, 0.9-50.1). N=29 received induction and consolidation entirely on an outpatient basis, ulterior hospitalizations during therapy were rare. Treatment abandonment was common (n=24; 26.4%) usually during induction (n=8; 32%) or consolidation (n=12; 48%) and mostly related to unaffordability. For the same reason, transfers to social security healthcare systems were also frequent (n=19; 20.9%). Most patients assessed were MRD negative (n=38; 74.5%) Early relapse incidence was 32.9%; 44.4% in MRD-positive and 27.5% in MRD-negative patients (p=0.43). OS at 24 months was 61.5% (95% CI 47-73%) and EFS 49.8% (95% CI 37-62%) with excellent outcomes for MRD-negative patients (Figure 1, Panels A and B). Treatment abandonment and MRD positivity were the only independent predictors of mortality in a multivariate analysis (HR 7.8 [95% CI 2.8-21.9] and HR 4.5 [95% CI 1.4-14], respectively). Lastly, the total cumulative price for medications included in our regimen was calculated at $16,750 vs. $36,061 USD in CALGB 10403, representing a cost reduction of 53.5%. Conclusions The treatment of Hispanic ALL patients with our regimen has shown promising outcomes at a reduced cost for patients. Genetic risk assessment, induction mortality, treatment abandonment and lack of access to novel therapies for MRD positive patients remain the main barriers for improving outcomes further. Figure 1 Figure 1. Disclosures Gomez-De Leon: Novartis: Honoraria; ASH: Research Funding; Sanofi: Honoraria; Abbvie: Honoraria. González López: JANSSEN: Honoraria; AMGEN: Honoraria. Gomez-Almaguer: Roche: Honoraria, Speakers Bureau; Janssen: Honoraria, Speakers Bureau; Bristol-Myers-Squibb: Honoraria, Speakers Bureau; Takeda: Honoraria, Speakers Bureau.

Background and objective Cytospin conventional cytomorphology (CCC) is the standard method for detecting lymphoblasts in cerebrospinal fluid (CSF) of patients with acute lymphoblastic leukemia (ALL) and for guiding treatment decisions. We evaluated flow cytometry immunophenotyping (FCI) performance for improving detection of central nervous system (CNS) involvement in ALL. Methods This prospective study included analysis of consecutive CSF samples of patients of all ages with ALL at 3 clinical stages: new diagnosis, relapse suspicion, and after relapse treatment. Manual, cytospin, automated, and FCI methods were compared and their performance statistically assessed. Using FCI as the reference method, optimal CSF cutoff cell count that better correlated with presence of lymphoblasts was established by receiver operating characteristic (ROC) curve analysis. Results Seventy-seven CSF samples were investigated, 35 (45.4%) from newly diagnosed cases, 30 (39%) suspicion of relapse, and 12 (15.6%) after treatment for relapse. Median manual WBC count in patients with CNS involvement detected by FCI was 3.75 cells/μL (0.0-1280), and this was also the count that best correlated with CNS infiltration (sensitivity, 50.0%; specificity, 82.2%). Compared with FCI, CCC sensitivity and specificity were 28.6% and 100%. Automated CSF WBC count in patients with CNS involvement detected by FCI was 5 (0.0-1578). For automated count, optimal WBC cutoff was 4.5 cells/μL (sensitivity, 62.5%; specificity, 70.5%). Conclusion Flow cytometry immunophenotyping complements conventional cytospin analysis for detection of lymphoblasts in the CSF of ALL patients at any clinical stage.

This article provides flow cytometry information regarding levels of expression for hematopoietic stem cell markers CD34 and CD133 obtained simultaneously of the bone marrow and peripheral blood from recipients of allogeneic and autologous transplants of PB hematoprogenitors for treating hematological malignancies and who were clinically healthy after ≥100 days following the procedure. CD34 and CD133 expression is compared regarding type of transplant (autologous vs. allogeneic) and sample cell source (bone marrow vs. peripheral blood). Patients were conditioned with a reduced-intensity conditioning regimen. Also shown is the flow cytometry analysis of mononuclear cell and lymphocyte populations in the peripheral blood of both types of recipients, as well as the characterization of immune cells, including T lymphocyte antigenic make up markers CD3, CD4 and CD8, B lymphocytes and NK cells, including total NK, bright and dim subtypes in the peripheral blood of both types of recipients. For further information and discussion regarding interpretation and meaning of post-transplant flow cytometry analysis, please refer to the article “Assessment of immune reconstitution status in recipients of a successful hematopoietic stem cell transplant from peripheral blood after reduced intensity conditioning” [1]. Keywords: Bone Marrow, CD34+ cells, CD133+ hematoprogenitors, Flow cytometry, Hematopoietic transplant, Peripheral blood, T, B and NK cells

Objective: By applying receiver operating characteristic curve analysis, the objective of this study was to see whether hemoglobin levels reflect body iron stores in a group of pregnant women at term who, by using serum ferritin as the reference test, had a high pre-test prob- ability of having iron deficiency anemia. Likewise, we evaluated the ability of hemoglobin and maternal serum ferritin levels to predict iron deficiency anemia in newborns. Methods: Hemoglobin and serum ferritin were measured in 187 pregnant women at term belonging to a group with a high pre-test probability of iron deficiency anemia and their newborns. Women with Hb

Objective To document immune reconstitution status after hematopoietic stem cell transplantation (HSCT) for malignant hematologic diseases. Methods Hematology patients who received a reduced intensity conditioning (RIC) were followed after successful allogeneic or autologous HSCT. Patients had at least 100 days post-transplant. T, B and NK cells in peripheral blood (PB), and CD34 +, CD133 + progenitor cells in bone marrow (BM) and peripheral blood (PB) were determined by flow cytometry. Results Twenty-seven HSCT recipients, 19 allogeneic and 8 autologous, were studied at a median 155 (100–721) days post-transplant. In the whole group the median value of CD34 + cells was 1.03% in the bone marrow and 0.04% in PB, whereas values for CD133 + cells were 0.39% and 0.13%, respectively, without statistical differences between autologous and allogeneic recipients. Significantly more B cells (CD3 −/CD56 −/CD19 +) were found in the autologous compared to the allogeneic group, 12.6 vs. 5.01, p = 0.04. An increased number of CD8 + lymphocytes with a 0.63 CD4:CD8 relationship was documented in PB. Conclusion In clinically recovered autologous and allogeneic HSCT recipients BM and PB CD34 +/CD133 + hematoprogenitor homeostasis is maintained within normal ranges, with better B-cell reconstitution in the autologous group.

Objective and background There is little information regarding the serologic status of umbilical cord blood (UCB) donors. Cytomegalovirus (CMV) is the most frequent agent transmitted by blood products and studies have reported that CMV can inhibit myelopoiesis, however, its effects on the cellular content of UCB have not been documented. Study design and methods We investigated, retrospectively, the prevalence of serological evidence of infection in 857 women donating their UCB at a public university hospital and studied the influence of acute CMV exposure on UCB content of CD34+ cells. The biological characteristics of UCB from serology positive-donors were compared with those of women with negative tests. Results We found that 51 of 857 (6%) UCB units were positive for infectious disease markers; anti-CMV IgM was the most prevalent marker, 43 of 51 (86%) of cases with infectious markers. UCB collected from anti‐CMV IgM-positive donors more frequently met rejection criteria for use as a transplanation product. The CD34+ cell count was the most often affected, 2.48 × 10 6 in anti‐CMV IgM-positive donors compared to 1.48 × 10 6 in unaffecetd donors( p = 0.006). The probability of a UCB meeting a CD34+ cell content ≥ 2 × 10 6 was significantly lower in units from IgM anti-CMV+ women compared to unaffecetd donors [Odds ratio (OR) = 0.428 (95% CI 0.182–0.632; p = 0.015]; the total nucleated cell count (TNC) was lower but not statistically significant [p = 0.068]. Conclusion UCB donated by anti-CMV IgM-positive women has a high probability of not meeting the criteria required for cryopreservation for future use as a transplantation product, because of the low number of CD34+ cells.

BACKGROUND: Infusion of an adequate dose of CD34+ mononuclear hematopoietic stem cells (HSCs) is the single most important variable to assure success in hematopoietic grafting. CD133+ HSCs constitute the CD34+ subgroup with higher differentiation potential. The number of granulocyte–colony-stimulating factor (G-CSF)-mobilized CD133+ HSCs administered during hematopoietic grafting and its relationship with the number of days needed to regain hematopoiesis was determined. STUDY DESIGN AND METHODS: Thirty-eight patients with malignant hematologic diseases who received an autologous (n = 15) or allogeneic (n = 23) HSC transplant were prospectively evaluated. G-CSF was administered for 5 days at 10 µg/kg/day. Hematopoietic progenitors were recovered from peripheral blood on day 5 by leukopheresis. CD34+ and CD133+/CD34+ cell populations were quantified by flow cytometry; the number of days to hematologic recovery was documented. RESULTS: A median dose of 4.56 × 106/kg CD34+ HSCs (range, 1.35 × 106-14.6 × 106) was recovered and transplanted; of these grafted cells, a median 3.25 × 106 were also CD133+ (range, 1.25 × 106-14.3 × 106). In the autologous group, the median number of days to reach a platelet (PLT) count of 20 × 109/L or greater was 12, and 15 days to obtain a neutrophil count of 0.5 × 109/L or greater; in the allogeneic group 13 and 16 days, respectively, were required (p > 0.05). A median 76.5% of G-CSF–mobilized CD34+ HSCs coexpressed the CD133+ antigen (range, 23.1-97.9). CONCLUSIONS: A higher number of CD133+/CD34+ HSCs in the graft was not clearly associated with a shorter neutrophil or PLT recovery time in either allogeneic or autologous recipients.

The regulatory proteins CD55 and CD59 are glycolsylphosphatidylinositol-anchored, type I cell surface proteins, which inhibit formation of the C3 convertases and prevent the terminal polymerization of the membrane attack complexes, respectively. Paroxysmal nocturnal hemoglobinuria (PNH) is a genetic disorder due to the impaired conformation of the glycolsylphosphatidylinositol anchor, which results in the deficient expression of CD55 and CD59 leading to excessive destruction of red cells and leukocytes. We have studied the expression of these two molecules in red blood cells, granulocytes and platelets in patients with PNH (two patients), autoimmune hemolytic anemia (AIHA) (seven patients), autoimmune thrombocytopenia (ATP) (22 patients), systemic lupus erythematosus (SLE) (19 patients), aplastic anemia (AA) (eight patients), and Evans syndrome (ES) (two patients). A diminished expression of CD55 and CD59 was found in the three cell lines of the two patients with PNH. In the seven patients with AIHA two were found with CD59 diminished expression in red blood cells and one with CD59 diminished expression in granulocytes. In the patients with ATP one was found with CD55 diminished expression in red blood cells, one with CD59 diminished expression granulocytes and one with a CD59 diminished expression in the platelets. In the subset of patients with SLE only one was found with a CD55 diminished expression in the red blood cells. In the patients with AA, a diminished expression in red blood cells was not found; however, one patient was found with a diminished expression of CD59 in granulocytes, and one patient with a diminished expression of CD55 in the platelets. In the two patients with ES we did not found changes in the expression of CD55 and CD59. We conclude that the diminished expression of the glycolsylphosphatidylinositol-anchored type I cell surface proteins CD55 and CD59 is not specific to PNH and that it can be found in patients with a variety of autoimmune disorders. Additional studies are needed to define the role of the deficiencies of CD55 and CD59 in the pathogenesis of autoimmune hemocytopenias.

BACKGROUND: Quantification of CD34+ mononuclear cells is the most important quality control measure for hematopoietic stem cell (HSC) transplantation. A fraction of CD34+ cells also express the CD133 antigen. These cells constitute a group of earlier, less-differentiated HSCs with a potentially higher capacity for engraftment. The correlation between total CD34+ peripheral HSCs and the fraction of these cells that coexpress CD133 was determined before and after automated collection by leukapheresis, as well as the effect of HSC CD133+ dose on hematopoiesis recovery. STUDY DESIGN AND METHODS: Granulocyte–colony-stimulating factor mobilization of HSCs from the marrow to the peripheral blood (PB) of allogeneic and autologous donors was followed by automated collection through leukapheresis on the fifth day. Quantification of CD34+ and CD133+ cells was performed on PB before collection and in the hematopoietic graft (HG) by flow cytometry. RESULTS: There was a significant correlation between CD133+ and CD34+ HSCs in the PB before collection and in the final product for grafting (r = 0.62 and 0.64; p