Xue M

Clinical analysis of the treatment of spinal cord injury with umbilical cord mesenchymal stem cells.

BACKGROUND AIMS The purpose of this study was to observe the clinical effect and safety of umbilical cord mesenchymal stem cells (UC-MSCs) in treating spinal cord injury (SCI) by intrathecal injection. METHODS From January 2008 to October 2010, we treated 22 patients with SCI with UC-MSCs by intrathecal injection; dosage was 1 × 10(6) cells/kg body weight once a week given four times as a course. Four patients received two courses, one patient received three courses and all other patients received one course. American Spinal Injury Association scoring system and International Association of Neurorestoratology Spinal Cord Injury Functional Rating Scale were used to evaluate neural function and ability to perform activities of daily living. RESULTS Treatment was effective in 13 of 22 patients; nine patients had no response. Among patients with incomplete SCI, the response to treatment was 81.25%; there was no response to treatment among six patients with complete SCI. Five patients with a response to treatment received two to three courses of therapy, and effects in these patients were further enhanced. In most patients in whom treatment was effective, motor or sensory functions, or both, were improved, and bowel and bladder control ability was improved. In 22 patients 1 month after therapy, algesia, tactile sensation, motion and activity of daily living scale were significantly improved (P < 0.01). During therapy, common adverse effects were headache (one case) and low back pain (one cases); these disappeared within 1-3 days. No treatment-related adverse events occurred during a follow-up period ranging from 3 months to 3 years. CONCLUSIONS UC-MSC therapy by intrathecal injection is safe and can improve neurologic function and quality of life in most patients with incomplete SCI.

Humanized anti-CD25 monoclonal antibody for prophylaxis of graft-vs-host disease (GVHD) in haploidentical bone marrow transplantation without ex vivo T-cell depletion

OBJECTIVE To investigate the effects of a novel anti-IL-2 receptor (CD25) monoclonal antibody, basiliximab, on graft-vs-host disease (GVHD) and engraftment in haploidentical bone marrow transplantation (BMT). MATERIALS AND METHODS Thirteen consecutive high-risk leukemia patients (age 9-41) underwent haploidentical BMT with G-CSF-primed marrow as stem cells without ex vivo T-cell depletion. Basiliximab, along with a combination of cyclosporine (CSA), methotrexate (MTX), and mycophenolate mofetil (MMF), was used for GVHD prophylaxis. Immunophenotyping, limited-dilution assay, and colony-forming assays were used to measure the effect of basiliximab on the subsets of lymphocytes, cytotoxic T-lymphocyte precursors (CTLp), and hematopoietic cells. RESULTS All patients established successful trilineage engraftment with full donor chimerism. No patients developed grade II-IV acute GVHD. Patients who survived more than 12 months and were free of relapse showed limited chronic skin GVHD. Ten of 13 patients are currently alive with a Karnofsky performance score of 100% at median follow-up of 17 months (range 12-24 months). Basiliximab significantly decreased alloreactive CTLp by 10-fold to 100-fold in limiting-dilution assays. It had no effect on hematopoietic stem and progenitor cells as determined by in vitro colony-forming assays. CONCLUSION The addition of basiliximab to CSA, MMF, and MTX as GVHD prophylaxis effectively reduced severe lethal GVHD in haploidentical BMT. It is possible to selectively eliminate or reduce the number of alloreactive T cells with anti-CD25 antibody, which results in prevention of or a reduction in the severity of GVHD.

HAPLOIDENTICAL HEMATOPOIETIC STEM CELL TRANSPLANTATION IN CHILD HEMATOLOGIC MALIGNANCIES WITH G-CSF-MOBILIZED MARROW GRAFTS WITHOUT T-CELL DEPLETION: A Single-Center Report of 45 Cases

In this report, the authors describe a protocol for haploidentical bone marrow transplantation in children who received G-CSF-mobilized bone marrow grafts without T-cell depletion from HLA-mismatched parents. Forty-two of 45 patients achieved complete donor hematopoietic engraftment; the medium time for neutrophil and platelet recovery was 17 and 19 days, respectively. Three died of early transplantation-associated complications; other causes of death included relapse (11 cases), fungal pneumonia (5), and acute graft-versus-host disease (2). The total disease-free survival rate longer than 2 years was 53.3%. These data suggest that haploidentical hematopoietic transplantation is an alterative strategy for children who lack immediate access to HLA-matched sources.

Haploidentical hematopoietic stem cell transplantation in hematologic malignancies with G-CSF mobilized bone marrow plus peripheral blood stem cells grafts without T cell depletion: a single center report of 29 cases

Abstract Haploidentical Hematopoietic stem cell transplantation (Haplo-HSCT) has provided an alternative option since virtually all patients have an immediately available donor. Here, we report the results of Haplo-HSCT with granulocyte-colony-stimulating factor (G-CSF) mobilized bone marrow grafts plus peripheral blood stem cells as the grafts without T-cell depletion. Twenty-nine patients with the mean age of 27.27 years (ranging from 15 to 51 years) were enrolled in this study, and 10 cases were in high risk status. The patients received myeloablative preconditioning with or without total body irradiation and acute graft-versus-host disease (GVHD) prophylaxis consisting of basiliximab, cyclosporine A, methotrexate, mycophenolate mofetil and a rabbit anti-thymocyte globulin. All the patients attained successful neutrophil and platelet recovery. The mean times for neutrophil and platelet recovery were 17.1 and 20.9 days, respectively. During the follow-up at a median time of 30.69 months (ranging from 3 to 76 months), nine patients developed aGVHD grade II–IV, including two developed grade III–IV GVHD after donor lymphocyte infusion. The incidence of cGVHD was 48.3%. 13 patients died within the first two years after transplantation, and the total disease-free survival rate longer than 2 years was 55.2%. These results suggest that G-CSF-primed bone marrow plus peripheral blood stem cell grafts are an appropriate stem cell source for Haplo-HSCT and large scale investigations are needed to confirm this protocol.

Co-transfusion of haplo-identical hematopoietic and mesenchymal stromal cells to treat a patient with severe aplastic.

A 3-year-old girl with severe aplastic anemia (SAA) that was unresponsive to steroid, cyclosporine and filgrastim treatments received bone marrow (BM) mesenchymal stromal cells (MSC; 1.25 x 10(6)/kg), granulocyte colony-stimulating factor (G-CSF)-mobilized BM and peripheral blood stem cell grafts from her father. Prior to stem cell transplantation, she had experienced repeated bacterial infections and received 44 blood transfusions during 8 months after diagnosis. The conditioning regimen consisted of fludarabine, cyclophosphamide and busulfan, and prophylaxis of acute graft-versus-host disease (GvHD) was performed by administration of anti-CD25 monoclonal antibody, cyclosporine A, methotrexate, mycophenolate mofetil and anti-thymocyte globulin. The patient achieved rapid hematopoietic engraftment of donor origin and no acute or chronic GvHD was observed. She is now alive with a good performance status, and the dose of cyclosporine A is being tapered. The novel regimen described here might be a suitable option for children with SAA who lack immediate access to HLA-matched sources.

Haploidentical hematopoietic stem-cell transplantation for non-Hodgkin lymphoma with bone marrow involvement

Here, we report the preliminary results of haploidentical hematopoietic stem cell transplantation (Haplo-HSCT) with granulocyte-colony-stimulating factor (G-CSF) mobilized bone marrow grafts without T-cell depletion for 10 patients with refractory non-Hodgkin lymphoma accompanied by bone marrow involvement. Eight patients received a conditioning regimen consisting of high-doses of cytarabine and cyclophosphamide with total body irradiation, whereas two cases were preconditioned with busulfan, thiotepa, and cyclophosphamide. All patients had rapid hematopoietic engraftment with the mean time for neutrophil and platelet recovery being 16.6 days and 19.2 days, respectively. Three cases died within 6 months after transplantation from severe acute graft-versus-host disease, fungal infection, or relapse. The others are currently alive in complete remission at a median follow-up of 60.71 months (range: 44–81months). The results here suggest that haplo-HSCT might provide an opportunity of myeloablative therapy for refractory lymphoma with marrow infiltration.

Functional mesenchymal stem cells remain present in bone marrow microenvironment of patients with leukemia post-allogeneic hematopoietic stem cell transplant

Abstract Mesenchymal stem cells (MSCs) and their progenies are important supporting cells in the bone marrow (BM) microenvironment. However, the function and kinetics of MSCs post-hematopoietic stem cell transplant (HSCT) remain unknown. In the present study, MSCs were cultured from a total of 76 BM samples from 15 patients receiving HSCT. Colony-forming unit fibroblasts in BM before pre-conditioning and 1, 3, 6 and 9 months post-HSCT were cultured and counted to quantify MSCs. Hematopoiesis-supporting activity of MSCs was observed with long-term culture of hematopoietic progenitors. An inhibitory effect of MSCs on in vitro lymphocyte proliferation was also observed. Results showed that post-HSCT MSCs supported in vitro hematopoiesis and inhibited lymphocyte growth. Moreover, the quantity of MSCs was reduced at an early stage and restored to baseline level 9 months post-transplant. The results indicate that functional MSCs remain present in the BM microenvironment, and these findings shed light on the understanding of BM microenvironment reconstitution post-HSCT.

Co-infusion of mesenchymal stromal cells has no effect on relapse and infection in patients with leukemia undergoing haploidentical hematopoietic stem cell transplant

We observed the outcome of co-transplant of umbilical cord-derived, ex vivo-expanded mesenchymal stromal cells (MSCs) in patients with leukemia undergoing haploidentical hematopoietic stem cell transplant (haplo-HSCT), and compared the relapse and infection rates with those in patients receiving traditional haplo-HSCT. The procedure was intended to assess the influence of MSCs on the relapse and infection rates of patients with leukemia after HSCT. From February 2010 to December 2012, 41 patients with leukemia received co-transplant of haplo-HSCT with MSCs (MSC  HSCT group). They were followed up for 21 months (2–50 months). From June 2008 to March 2010, 42 patients with leukemia received traditional haplo-HSCT (HSCT group). They were followed up for 35 months (2–70 months). This study was approved by the ethics and technological committees of the General Hospital of the Air Force, and all patients provided informed consent before MSC use. We found that the mean time for neutrophil engraftment in the MSC  HSCT group was significantly shorter than in the HSCT group (16.90  0.63 days vs. 18.83  0.63 days, p  0.03). There was no significant difference in the cumulative incidence of II–IV acute graft versus host disease (aGVHD) or chronic GVHD (cGVHD) between MSC  HSCT and HSCT groups (p  0.109 and p  0.556, respectively). In addition, no notable effect of MSC co-infusion was found on overall survival (p  0.573), relapse-free survival (p  0.424) and event-free survival (p  0.222) or non-relapse mortality (p  0.853) in patients who underwent traditional haplo-HSCT. Also, there was no significant difference in the rate of leukemia relapse (p  0.736), pulmonary infection (p  0.182), cytomegalovirus (CMV) infection (p  0.652) or pneumonia-associated death (p  0.396) within 1 year after haplo-HSCT between MSC  HSCT and HSCT groups. Our results indicate that co-infusion of MSCs can shorten the time of neutrophil engraftment, but does not increase the rates of relapse and infection in haplo-HSCT. Eighty-three patients received a regimen consisting of busulfan at 4 mg/kg/day orally or 0.8 mg/kg/day intravenously (IV) (days  6,  7 and  8), cytarabine at 4 g/m2/ day (days  4 and  5) and cyclophosphamide at 55 mg/ kg/day (days  2 and  3). Stem cell collection and MSC preparation were performed as previously reported [1,2]. The MSCs were passaged, and cells at passage 3 were used for therapy after phenotypic analysis with fluorescence activated cell sorting (FACS) (FACSCalibur; BD, Franklin Lakes, NJ) (CD44, CD73, CD166, CD45, HLA-DR, CD14, CD31). At day 0, under monitoring of vital signs, patients were given MSCs IV via a central venous catheter before hematopoietic graft infusion. GVHD prophylaxis was conducted using a combination of immunosuppressive drugs including cyclosporine A (CSA), methotrexate (MTX), mycophenolate mofetil (MMF) and rabbit anti-thymocyte globulin (ATG; Fresenius AG, Oberursel, Germany). Briefly, before transplant, CSA was administered IV at 1.5 mg/kg/day from day  7 to day  2, and the dose was increased to 2.5 mg/kg/day from then until recovery of intestinal function, when it was given orally at 5 mg/kg/day. ATG was administered at a dose of 5 mg/kg/day from day  4 to day  1. MTX was administered IV at 15 mg/m2 on day  1 and at 10 mg/m2 on days  3,  6 and  11. MMF was administered orally at 0.25–0.5 g/day from day  7 to day  30 (Supplementary Figure 1, to be found at online http://informahealthcare. com/doi/abs/10.3109/10428194.2015.1020061). All patients were monitored for engraftment and post-transplant adverse events, including GVHD, relapse and infection. Hematopoietic stem cell engraftment was defined as maintenance of an absolute peripheral neutrophil count above 0.5  109/L or an unsupported platelet count above 20  109/L for 3 consecutive days. Hematopoietic chimerism was identified by blood type analysis (blood type mismatch), sex chromosome determination (sex mismatch) and/or polymerase chain reaction (PCR)-DNA fingerprinting (short tandem repeat) to confirm Leukemia & Lymphoma, October 2015; 56(10): 2965–2968