Immunoreconstitution after human bone marrow transplantation

Immunologic reconstitution was studied in 24 patients who underwent bone marrow transplantation, 17 allogenic and 7 autologous. The GVHD prophylaxis consisted of methotrexate and prednisone. The complete immune evaluation was to be carried out prior to transplantation at 1, 2, 3, 6, 9, 12 months after BMT and subsequently every 6 months up to 4 years. The investigated immunological parameters included total lymphocyte count, B-lymphocytes, T3-, T4-, T8-lymphocytes, T4/T8 ratio, natural killer cell activity, ADCC, lymphocyte blastogenic response and serum-IgG, -IgA, -IgM. Absolute lymphocyte count, B-lymphocytes, T3-lymphocytes recovered to normal levels after 6 months. T4-lymphocytes decreased significantly during the first 180 days posttransplant. T8-lymphocytes increased after 6 months to values higher than normal and the T4/T8 ratio decreased significantly and continued below 0.8 for 48 months. Patients without and with GVHD had low lymphocyte response to PHA and Con A for the first 6 months.     

Genetic markers in human bone marrow transplantation.

Blood cell isozymes, red cell antigens, immunoglobulin allotypes, and marker chromosomes are suitable tools to monitor bone marrow engraftment and marrow graft quality. Data on genetic markers from 26 patients who underwent bone marrow transplantation as a treatment for acute leukemia are presented here.     

Design of human granzyme B variants resistant to serpin B9

Human granzyme B (hGB) is a serine protease involved in immune‐mediated apoptosis. Its cytotoxicity makes it potentially applicable in cancer therapy. However, the effectiveness of hGB can be hampered by the cytosolic expression of a natural protein inhibitor, human Serpin B9 (hSB9). Here, we used computational approaches to identify hGB mutations that can affect its binding to hSB9 without significantly decreasing its catalytic efficiency. Alanine‐scanning calculations allowed us to identify residues of hGB important for the interaction with hSB9. Some variants were selected, and molecular dynamic simulations on the mutated hGB in complex with hSB9 in aqueous solution were carried out to investigate the effect of these variants on the stability of the complex. The R28K, R201A, and R201K mutants significantly destabilized the interaction of the protein with hSB9. Consistently, all of these variants also retained their activity in the presence of the Serpin B9 inhibitor in subsequent in vitro assays of wild‐type and mutated hGB. In particular, the activity of R201K hGB with and without Serpin B9 is very similar to that of the wild‐type protein. Hence, R201K hGB emerges as a promising species for antitumoral therapy applications. Proteins 2012. © 2012 Wiley Periodicals, Inc.     

An overview of bone marrow transplantation for chronic myeloid leukemia.

The use of intensive therapy together with transplantation of marrow from a suitable donor is the only established curative treatment for patients with chronic myeloid leukemia (CML). However, marrow transplantation is hazardous, costly and applicable to relatively few patients. Therefore, we evaluated the results and limitations of marrow transplantation for CML and discussed new treatment strategies. We decided to select a limited number of papers that focused on the relevant issues rather than to undertake an exhaustive comparison of treatment results from different centres. Patients with CML in the chronic phase who receive marrow from a sibling with the same human leukocyte antigen type can expect to have a long-term disease-free survival rate of 50%. However, the procedure is associated with a mortality rate of 30%, mainly because of graft-versus-host disease (GVHD) and interstitial pneumonitis. Moreover, because of the requirements for age and histocompatibility only 10% of patients with chronic-phase CML are currently eligible. Transplantation earlier in the chronic phase (within 1 year after diagnosis), the use of marrow from matched, unrelated donors and the development of improved methods for reducing the incidence of GVHD all hold promise. In addition, the preliminary results of intensive therapy followed by transplantation with cultured autologous marrow have been encouraging. If further progress is to be made, continued optimism coupled with carefully developed and executed studies will be necessary.     

HLA compatibility for patients with thalassemia: implications for bone marrow transplantation

In order to evaluate the possibility of a patient with thalassemia finding an HLA-identical sibling donor, we performed an analysis of HLA antigens in families of thalassemic patients. The pattern distribution was not significantly different from the expected ratio 25:50:25. When the siblings were subdivided according to the age of the patients (under or over 5 years), the above pattern remained unmodified for both the age groups. The average size of the 129 thalassemic families was 2.4. Thus, taking into account that thalassemic patients have an average of 1.4 siblings and that the HLA genotype distribution is expected as such, HLA-matched siblings are available for 33% of the patients. Because an additional 8.5% of the patients were found HLA-phenotypically identical to one parent, the chance for a patient with thalassemia to find a suitable donor for bone marrow transplantation would be increased to 41.5%. Our preliminary data cannot be extrapolated to the overall Italian thalassemic population; however, it can be inferred that for a patient with thalassemia, the chance of finding a suitable donor for bone marrow transplantation is not reduced.     

Advantages of peripheral blood stem cells from unrelated donors versus bone marrow transplants in outcomes of adult acute myeloid leukemia patients

Background aimsIn allogeneic stem cell transplantation, unrelated donors are chosen in cases where appropriate related donors are not available. Peripheral blood stem cells (PBSCs) are more often selected as a graft source than bone marrow (BM). However, the prognostic benefits of PBSCs versus BM transplants from unrelated donors have not been carefully examined in patients with acute myeloid leukemia (AML). This study compared outcomes of adult AML patients who underwent unrelated PBSC and BM transplantation, evaluating post-transplant complications, including engraftment, graft-versus-host disease (GVHD) and infections, and determined subgroups of patients who are most likely to benefit from unrelated PBSCs compared with BM transplants.MethodsThe authors analyzed 2962 adult AML patients who underwent unrelated PBSC or BM transplants between 2011 and 2018 (221 PBSC and 2741 BM) using the Japanese nationwide registry database, in which graft source selection is not skewed toward PBSCs.ResultsIn 49.7% of patients, disease status at transplantation was first complete remission (CR1). In 57.1% of cases, HLA-matched donors were selected. Myeloablative conditioning was performed in 75.1% of cases, and anti-thymocyte globulin (ATG) was added to conditioning in 10.5%. Multivariate analyses showed a trend toward favorable non-relapse mortality (NRM) in PBSC recipients compared with BM recipients (hazard ratio [HR], 0.731, P = 0.096), whereas overall survival (OS) (HR, 0.959, P = 0.230) and disease-free survival (DFS) (HR, 0.868, P = 0.221) were comparable between PBSC and BM recipients. Although the rate of chronic GVHD (cGVHD) was significantly higher in PBSC patients (HR, 1.367, P = 0.016), NRM was not increased, mainly as a result of significantly reduced risk of bacterial infections (HR, 0.618, P = 0.010), reflecting more prompt engraftments in PBSC recipients. Subgroup analyses revealed that PBSC transplantation was advantageous in patients transplanted at CR1 and in those without ATG use. PBSC recipients experienced significantly better OS and/or DFS compared with BM recipients in this patient group.ConclusionsThe authors' results confirmed the overall safety of unrelated PBSC transplantation for adult AML patients and suggested an advantage of PBSCs, especially for those in CR1. Further optimization of the prophylactic strategy for cGVHD is required to improve the overall outcome in transplantation from unrelated PBSC donors.     

Immunoperoxidase detection of myeloid antigens in glycolmethacrylate-embedded human bone marrow

Different methods for fixation and exposure of antigenic determinants were tested for detection of a granulocytic differentiation antigen by the monoclonal antibody L12-2, using an indirect immunoperoxidase method on semi-thin sections of undecalcified, glycolmethacrylate-embedded human bone marrow biopsies. Fixation in Bouin's solution for 3 hr gave a more intense and more homogeneous immunological staining than fixation in absolute methanol, 4% formalin, B5, or Michel's medium, and the morphological detail was excellent. Digestion by pronase or trypsin was required. Coating the glass slides with Alcian blue prevented loss of sections from the slides during the staining procedure. Bouin fixation also made possible detection of two other differentiation antigens expressed in the granulocytic series, using the monoclonal antibodies 1G10 and R1B19. Furthermore, the same technique also permitted detection of factor VIII-RAg in the megakaryocytes, as well as recognition of cells of the erythroid series by use of polyclonal rabbit antisera.     

Regression of syndesmophyte after bone marrow transplantation for acute myeloid leukemia in a patient with ankylosing spondylitis: a case report

ABSTRACT: INTRODUCTION: Disease progression of ankylosing spondylitis has been considered irreversible. However, wereport a case of spontaneous regression of syndesmophyte development following allogeneicperipheral blood stem cell transplantation in a patient with acute myeloid leukemia, who wasalso diagnosed as having ankylosing spondylitis. To the best of our knowledge, this is thefirst case report presenting the partial radiologic regression of syndesmophytes. CASE PRESENTATION: A 39-year-old man with active ankylosing spondylitis achieved clinical remission and partialradiological regression of cervical spine syndesmophytes following a peripheral blood stemcell transplantation for acute myeloid leukemia. Our patient received an allogeneic peripheralblood stem cell transplantation following a pre-transplantation conditioning regimen of totalbody irradiation and cyclophosphamide. The donor was a human leukocyte antigen-matched29-year-old man. Our patient has remained asymptomatic and has received no medication forankylosing spondylitis for nearly three years. CONCLUSIONS: Several explanations are proposed for the regression of syndesmophytes and clinicalimprovement in active ankylosing spondylitis observed in our patient, including changes inbone remodeling and immune reconstitution following stem cell transplantation, the effect ofimmunosuppressive agents, or fluctuation in the natural course of ankylosing spondylitisalthough further studies are required. The regression of syndesmophytes in ankylosingspondylitis in this case raises the possibility that stem cell transplantation might contribute tothe development of a novel therapeutic strategy for treatment of the disease.     

HSP10 selective preference for myeloid and megakaryocytic precursors in normal human bone marrow

Heat shock proteins (HSPs) constitute a heterogeneous family of proteins involved in cell homeostasis. During cell life they are involved in harmful insults, as well as in immune and inflammatory reactions. It is known that they regulate gene expression, and cell proliferation, differentiation and death. HSP60 is a mitochondrial chaperonin, highly preserved during evolution, responsible of protein folding. Its function is strictly dependent on HSP10 in both prokaryotic and eukaryotic elements. We investigated the presence and the expression of HSP60 and HSP10 in a series of 20 normal human bone marrow specimens (NHBM) by the means of immunohistochemistry. NHBM showed no expression of HSP60, probably due to its being below the detectable threshold, as already demonstrated in other normal human tissues. By contrast, HSP10 showed a selective positivity for myeloid and megakaryocytic lineages. The positivity was restricted to precursor cells, while mature elements were constantly negative. We postulate that HSP10 plays a role in bone marrow cell differentiation other than being a mitochondrial co-chaperonin. The present data emphasize the role of HSP10 during cellular homeostasis and encourage further investigations in this field.     

针对骨髓移植病患中铁过载的去铁治疗研究进展

骨髓移植是临床上治疗恶性造血系统疾病的常见手段。而铁过载是临床上常见的并发症之一,对病患的造血功能和治疗后恢复有极大的抑制作用。了解铁过载产生的分子或遗传机制能帮助优化去铁化方案,提高去铁化治疗的效率。本文总结了骨髓移植前后铁过载现象发生机制的最新研究进展,并阐述了临床上多种去铁治疗的方案,以期为该类病患铁过载的预防和治疗提供参考。     

Cytokine activity after human bone marrow transplantation. Production of interferons by peripheral blood mononuclear cells from recipients of HLA-identical sibling bone marrow transplants

Peripheral blood mononuclear cells (PBMC) from 43 recipients of HLA-identical sibling bone marrow transplants were tested for their capacity to produce IFN after stimulation in vitro with PHA. Total IFN production, as measured in a bioassay, and production of IFN-gamma, measured by radioimmunoassay, was within or above the normal range (mean production by PBMC from normal persons +/- 2 SD) for 29 of 39 and 31 of 43 recipients, respectively. Production did not correlate with time after transplant, or with the presence or type of immunosuppressive chemotherapy used, although there was a tendency for lower production in recipients with chronic graft-vs-host disease. This near normal production of IFN contrasts with our previous observation that IL 2 production in bone marrow transplant recipients is severely impaired for at least 6 mo post-transplant.