Flow cytometric monitoring of minimal residual diseases in patients with acute leukemia after allogeneic hemapoietic stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the significance of flow cytometric monitoring minimal residual diseases (MRD) in patients with acute leukemia (AL) after allogeneic hemapoietic stem cell transplantation (HSCT).</p><p><b>METHODS</b>From January 2007 and January 2008 MRD were detected by flow cytometry (FCM) in 402 bone marrow (BM) in 102 AL patients without leukemic gene and chromosomal changes at first diagnosis after HSCT (1, 2, 3, 6,12 months after HSCT; adding detection frequency in part of high risk patients), The relationship between the MRD results and clinical prognosis were observed. Patients with significantly higher MRD were treated and the effectiveness was monitored by FCM (MRD > 0.01% considered as positive).</p><p><b>RESULTS</b>(1) 71 cases were persistently negative for MRD after HSCT and all them were in hematologic complete remission (CR). Only 3 cases had extramedullary relapse. The disease free survival (DFS) and overall survival (OS) were 66.2% and 90.1%, respectively. (2) Of 27 MRD(+) cases 11 converted to MRD negativity after chemotherapy plus donor lymphocyte infusion (DLI), CIK, NK cells. The DFS and OS were 63.6% and 72.7%, respectively. Other 16 cases had hematologic relapse. The DFS and OS were 11.1% and 25.0%, respectively. The median time from MRD increasing to hematologic relapse was 48 days (7-69 day). (3) Four cases had hematologic relapse after HSCT and died in the end.</p><p><b>CONCLUSIONS</b>(1) The DFS and the OS in MRD(-) cases are significantly higher than those of MRD(+) cases. (2)MRD(+) patients after HSCT coveted to MRD(-) after intervention. Therapy, whose DFS and the OS are still significantly higher than those of MRD(+) cases. (3) Patients with hematologic relapse after HSCT have the worst prognosis and the DFS and OS are significantly low. FCM monitoring of MRD in patients after HSCT is a sensitive, specific, quick and simple method. It can indicate recurrent state in time, facilitates early intervention, reduces the hematologic relapse risk and improves DFS.</p>

Veto T cells enhance engraftment allogeneic hematopoietic stem cells / 中国实验血液学杂志

Veto activity was defined as the capacity of specifically downregulating cytotoxic T-precursor cell (CTL-p) directed against antigens of the veto cells themselves but not against third-party antigens. Many studies have shown that the most potent veto cells are CD8(+) cytotoxic T lymphocytes (CD8(+)CTLs). Effectively, CD8(+)CTLs of donor origin can facilitate engraftment of donor's stem cells by eliminating host-alloreactive T lymphocytes. In this article, effect mechanisms, depletion of GVH ex vivo, application in vivo, synergistic enhancement with rapamycin and regulatory T cells, and anti-tumor effect in the hematopoietic stem cell transplantation are summarized.

Cytomegalovirus enteritis after allogeneic hematopoietic stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyse the clinical features, diagnostic methods and risk factors of cytomegalovirus (CMV) enteritis after allogeneic hematopoietic stem cell transplantation (allo-HSCT).</p><p><b>METHODS</b>Analysis was made on 24 cases of CMV enteritis after allo-HSCT in Beijing Daopei Hospital from Aug. 2007 to Jul. 2009, including clinical data, endoscopic diagnosis, histopathological and virological results, and the association between CMV enteritis with viremia and graft-versus-host disease(GVHD).</p><p><b>RESULTS</b>87.5% of the patients were over 18 years old. The median time to diagnosis of CMV enteritis was 63 days after HSCT. The mucosal lesions in enteroscopic examination had no significant differences between CMV enteritis and gastrointestinal GVHD complicated with the enteritis. The methods used in diagnosis included histopathology (32.1%) and virology (92.9%). The copies of CMVDNA in mucosal samples greater than 10(5)/10(6) PBNC was better diagnosis. A number of risk factors were compared between the survival and death groups: type of transplant, conditioning regimen, the time span of ganciclovir prophylaxis therapy, grade II-IV GVHD before enteritis, the time of diagnosis as GVHD, using MP > or = 1 mg/kg to treat GVHD, the time between GVHD and enteritis, CMV viremia before enteritis, the time of diagnosis as enteritis, CMVDNA quantitation, and there were no any statistic differences.</p><p><b>CONCLUSION</b>Cytomegalovirus enteritis should be carefully diagnosed by histopathology and virology through endoscopic examination. It is better to undertake pan-colon endoscopy as well as terminal ileum examination for more accurate diagnosis. PCR can significantly improve the detection rate. CMVDNA detection in patients' stool may be helpful to diagnosis, especially for those patients who can not stand the endoscopy examination.</p>

Clinical research on reconstitution of CD4+CD25+ T cells after haploidentical bone marrow transplantation / 中国实验血液学杂志

The aim of this study was to investigate the reconstitution of CD4(+)CD25(+) T cells after haplo-identical bone marrow transplantation (hiBMT) and its correlation with graft versus host disease (GVHD) and relapse. Peripheral blood samples from 27 patients after hiBMT were harvested and the percentage and absolute counts of CD4(+)CD25(+) T cells were detected by flow cytometry. The correlations of GVHD occurrence and disease relapse with the reconstitution of CD4(+)CD25(+) T cells were analyzed. The results showed that the percentage of CD4(+)CD25(+) T cells of peripheral blood samples increased significantly after G-CSF priming. At day 30 after hiBMT, CD4(+)CD25(+) T cells were recovered to the 20% of normal level, followed by a slowly process in 3 months, and up to one half of the normal level at 180 days. There was no evidence to prove relationship between CD4(+)CD25(+) T cells and acute GVHD, while CD4(+)CD25(+) T cells were increased significantly in the chronic GVHD group. The absolute count of CD4(+)CD25(+) T cells showed no relations with relapse of leukemia during the first year after hiBMT. In conclusions, chronic but not acute GVHD was in relation to the reconstitution of CD4(+)CD25(+) T cells based on the anti-CD25 antibody therapy model for the prevention of GVHD after hiBMT. Further investigation is needed to clarify whether the relapse of leukemia after hiBMT is related to the reconstitution of CD4(+)CD25(+) T cells.

Application of fibrotic bronchoscopy in the diagnosis of pulmonary diffuse infiltration following bone marrow transplantation / 中国实验血液学杂志

In order to evaluate the diagnostic value of fibrotic bronchoscopy (FB) in the pulmonary infiltration following bone marrow transplantation (BMT), 18 patients with pulmonary complications after BMT from November 2003 to March 2006 were performed with FB. Bronchoalveolar lavage (BAL) and brushing were performed in patients who had received short-term empirical therapy without good response, and transbronchial lung biopsy (TBLB) was carried out in 3 cases. The results showed that 9 out of 10 cases with pulmonary infection, including bacterial pneumonia (n = 3), aspergillosis (n = 2), pneumocystis carinii pneumonia (n = 3) and viral infection (n = 1) were diagnosed by using FB. One case was diagnosed as tuberculosis after open lung biopsy following negative results from twice BAL. 2 out of 8 cases were diagnosed by TBLB as noninfectious pulmonary complications. In conclusion, FB, especially with BAL, is a safe and useful procedure for the evaluation of pulmonary complications, which is particularly suitable for diagnosis of pulmonary infection after BMT. Furthermore, TBLB should be recommended in order to avoid open lung biopsy, if the patients tolerate the operation.

Correlation between CD34+CD61+ megakaryocyte precursors and platelet engraftment in allogeneic hematopoietic stem cell transplantation / 中国实验血液学杂志

This study was purposed to investigate the correlation between the dose infused megakaryocytic precursors (CD34+, CD34+CD61+) and recovery time of platelet count following an allogeneic PBSCT and/or BMT through quantitative detection of CD34+ and its subpopulation in peripheral blood and BM mobilized by G-CSF. 24 patients with various hematologic malignancies received PBSCT/BMT from their HLA matched or unrelated donors and haploidentical siblings in April-December 2007. 20 evaluated patients were divided into 2 groups according to different transplant schemes. HLA matched group received PBSCT regime and haploidentical group received PBSCT combined with BMT. CD34+CD61+ subpopulations in sample from patients receiving PBSCT/BMT were measured by flow cytometry immediately or storage over night. The results showed that the median number of infused CD34+, CD34+CD61+ and CD34-CD61+ cells in haploidentical group were 6.24x10(6)/kg (1.53-20.48), 66.19x10(4)/kg (8.16-493.83), and 34.38x10(6)/kg (14.71-109.16) respectively, in HLA matched group those were 4.88x10(6)/kg (1.00-8.24), 14.16x10(4)/kg (11.63-96.87), and 13.50x10(6)/kg (1.74-35.61), respectively. Median days of ANCs>0.5x10(9)/L and platelets>20x10(9)/L were 18.5 (11.0-29.0) days and 16.5 (9.0-35.0) days in haploidentical group respectively; in HLA matched group those were 14.5 (9.0-24.0) and 10.5 (6.0-37.0) respectively. A significance difference of median days for ANC engraftment presented between two groups (p=0.048). There was no significant difference of time for platelet engraftment between 2 groups. For patients with CD34+ cell dose>2x10(6)/kg there was significant difference of time of platelet engraftment between HLA matched and haploidentical groups (p=0.006). The number of CD34+CD61+ cells infused in 12 haploidentical patients or in 8 HLA matched patients were much better correlated with the time of platelet recovery up to 20x10(9)/L than that of number of CD34+ cells infused in total 20 patients (r=-0.768 and p=0.004 for haploidentical CD34+CD61+ cells, r=-0.747 and p=0.033 for HLA matched CD34+ CD61+ cells, r=-0.449 and p=0.047 for CD34+ cells). There was an inverse correlation between the number of infused CD34+ CD61+ cells and time of platelet engraftment. Therefore, as the number of CD34+ CD61+ cells increased, duration of platelet engraftment (time to reach platelet count of 20x10(9)/L) shortened significantly. It is concluded that the determining the number of megakaryocytic precursor by flow cytometry may predict the platelet reconstitutive capacity of the allogeneic hematopoietic stem cell transplantation, which is in haploidentical PBSCT and in BMT.

Clinical investigation on treatment of children leukemia with non-T-cell depleted haploidentical bone marrow transplantation / 中国实验血液学杂志

To investigate the efficacy and feasibility of parent non-T cell depleted haploidentical bone marrow transplants (haploidentical BMT) for children with leukemia, the efficacy of haploidentical BMT was evaluated in 8 leukemia children (1.9-9 years) received hematopoietic stem cell transplantation, donors were their parents with HLA-mismatched for two or three loci. Five children were pre-conditioned with a myeloablative regimen consisting of high-dose cytarabine (Ara-C), cyclophosphamide (CY) and total body irradiation. Busulfan (BU), Ara-C and CY were used for preconditioning regimen in other three children. The donors were given G-CSF prior to marrow harvest and the non-T-cell depleted grafts were used. A combination of CsA, MTX, ATG, mycophenolate mofetil (MMF) and CD25 monoclonal antibody were used for GVHD prophylaxis. The results showed that rapid engraftment was observed in all cases after transplantation by cytogenetic evidence. The mean time of neutrophil count exceeded 0.5 x 10(9)/L and the mean time of platelet count exceeded 20 x 10(9)/L were 16 and 17 days after transplantation respectively. Incidence of lethal aGVHD was lower, II-III acute aGVHD was found only in one out of eight patients. Chronic GVHD was observed in five patients, 4 from which showed local cGVHD, one developed extensive cGVHD. During the follow-up of 33 months (range 7-56 months), two patients died from relapsed leukemia, including one relapsed as donor-origin leukemia. Disease-free survival was achieved in the remaining six patients. No death occurred during the follow-up of 6 months. It is concluded that above-mentioned preconditioning regimen and GVHD prophylaxis procedure in non-T-cell depleted bone marrow transplantation from HLA-mismatched parents are effective approaches and safe strategy for the treatment of children leukemia.

Impact of incompatible killer cell immunoglobulin-like receptor and its ligand on the outcome of haploidentical bone marrow transplantation / 中国实验血液学杂志

The purpose of study was to investigate the impact of killer immunoglobulin-like receptor (KIR) and its ligand on haploidentical bone marrow transplantation. 74 cases were analyzed for the distribution frequencies and characteristics of KIR and its ligand as well as the impact of KIR ligand for the haploidentical bone marrow transplantation in terms of the overall survival, disease-free survival (DFS), GVHD and relapse. The results showed that among the 19 KIR genotypes currently nominated KIR2DL1, KIR2DL4 and KIR3DL2-3 could be detected in all the cases. Other high frequency genotypes included KIR3DP1 (98.6%), KIR2DP1 (98.6%), KIR3DL1 (97.3%) and KIR2DL3 (97.3%). Inhibitory receptor genotypes were 1.37-fold of activating receptor genotypes. KIR2DL1, KIR3DL2, KIR3DL3 and KIR2DL4 were found in all haplotypes and at least one genotype of KIR2DL2 and/or KIR2DL3 existed in all haplotypes. Among the 14 genotypes found in the test, the HLA-Cw7 was the most popular (37.8%) and the group 2 (HLA-Cw1, 3, 7, 8, 13, 14) recognized by KIR2DL2/2DL3 counted for 43.2%. The incompatibility of KIR for 32 cases of haploidentical BMT was 43.8%, of which 9/14 were KIR2DL incompatible, 5/14 were KIR2DL2 or KIR3DL1 incompatible. Among the 46 cases of haploidentical BMT, 29 cases were HLA-Cw matched and 14 cases were mismatched. The completed mismatch ratio of HLA-Cw was 30.4% and the match ratio was 63.4%. The survival rate was higher for the 14 cases of KIR genotype compatible group than the 13 cases of KIR genotype incompatible group (p = 0.032). The disease-free survival was significantly higher for the 17 cases of mismatched KIR ligands (HLA-Cw) group than the matched group (p = 0.024). The survival rate was higher in GVHD group than that in non-GVHD group when the KIR ligand was missing. The acute and severe GVHD was related to the existence of activating receptor of KIR2DS1/2DS2. The incompatibility group was accompanied with frequent acute and severe GVHD and less relapse and vice versa for the compatibility group. One patient died after BMT among the 14 mismatched KIR ligand group suffering from myelogenous leukemia while 4 patients out of 12 patients died in the matched group. It is concluded that the haploidentical BMT is characterized by mismatch between donor and recipient and its immunological reactions also features by the incompatibility of KIR genotype and missing ligand. The missing ligand for the donor KIR has strong effect on the outcome of BMT and it means a lot to analyze the KIR genotype and its ligand for the selection of best donor and prognostic evaluation in haploidentical BMT.

Antithymocyte globulin used for treatment of severe acute graft versus host disease after haploidentical bone marrow transplantation / 中国实验血液学杂志

The objective of study was to investigate the effect of low-dose antithymocyte globulin (ATG) on steroid-resistant severe acute graft versus host disease (aGVHD). Six patients with steroid-resistant severe aGVHD after haploidentical bone marrow transplantation (BMT) received the treatment with ATG at a low dose of 1.25 mg/kg for 3 - 5 doses every other day. The results showed that 3 out of 6 patients got completely remission (CR), among them 2 patients have still been in disease-free survival, 1 patient died from leukemia relapse. 1 out of the other 3 patients got partial remissin (PR), 2 patients were aggravated. The other 3 patients all died from GVHD. The major complications observed in these patients were infections. In conclusion, low-dose ATG is effective for some patients with steroid-resistant severe aGVHD, and has not severe side effect. To strengthen environmental protection should be considered as important for prevention of infection.

Neurological complications following haploidentical bone marrow transplantation / 中国实验血液学杂志

<p><b>UNLABELLED</b>To explore the occurrence patterns of neurological complications following haploidentical bone marrow transplantation, a series of clinical data as the onset time, etiology, choices of therapies and prognosis in 10 patients with neurological disorders were summarized. The results showed that complications occurred in 10 out of 74 patients after bone marrow transplantation, which include 3 with encephalorrhagia, 3 infection, 1 epilepsy, 1 Guillian-Barr's syndrome, 1 encephalopathy and 1 schizophrenia. 4 patients died of neurological complications.</p><p><b>IN CONCLUSION</b>neurological complications commonly occurred in haploidentical bone marrow transplantation, and encephalorrhagia might be the main indication that needs intensive care. Moreover, central nerve system infection and peripheral nerve diseases associated with slow immune reconstitution should have clinical interests.</p>

Pulmonary complications in haploidentical bone marrow transplantation / 中国实验血液学杂志

<p><b>UNLABELLED</b>To explore the occurrence patterns of pulmonary complications at different stages in haploidentical bone marrow transplantation, a series of clinical data as the onset time, etiology, management choices and prognosis in 18 patients with pulmonary disorders were summarized. The results showed that in 18 out of 70 patients after bone marrow transplantation occurred pulmonary complications which included pneumonia affected by bacteria (7 cases), fungus (5 cases) and cytomegalovirus (CMV, 4 cases), bronchiolitis obliterans organizing pneumonia (BOOP, 1 case), and idiopathic pneumonia syndrome (1 case), out of which 8 cases died. Fungal and CMV pneumonia occurred predominantly 2 to 3 months after transplantation, whereas bacterial pneumonia was observed in the duration of 3 to 12 months and 4 cases of them suffered from secondary fungal infections during treatment. BOOP and idiopathic pneumonia syndrome were diagnosed 12 months and 50 days after transplantation respectively.</p><p><b>IN CONCLUSION</b>pulmonary complications were commonly seen in haploidentcal bone marrow transplantation, and fungal pneumonia might be the main cause that needs intensive management.</p>

CD25 monoclonal antibody for GVHD prophylaxis in non-T-cell depleted haploidentical bone marrow transplantation for treatment of childhood leukemia / 中华儿科杂志

<p><b>OBJECTIVE</b>Allogeneic bone marrow transplantation (Allo-BMT) has improved long-term survival in children patients with refractory leukemia. For patients who do not have an HLA-identical sibling, the treatment option is limited. Using related mismatch donors or parental donors for Allo-BMT was at high risk for acute severe GVHD. The purpose of this study was to explore the effects of CD25 monoclonal antibody on reducing the incidence of severe acute GVHD in haploidentical bone marrow transplantation for the treatment of childhood leukemia.</p><p><b>METHODS</b>Ten children with leukemia received haplotype Allo-BMT from HLA two or three loci mismatched related donors. Most patients were classified as in high risk category. The donors of patients were given G-CSF (Lenograstim Chugai) 250 microg/day for seven doses prior to marrow harvest. The non-T-cell depleted haploidentical bone marrow was infused. In addition to combination of CSA, MTX, ATG (Fresenius Hemocare, Germany) and mycophenolate mofetil (MMF) for GVHD prophylaxis, CD(25) monoclonal antibody (Simulect, Novartis Pharma Switzerland) was administered to prevent acute severe GVHD.A total of 40 mg Simulect was given in two doses of 20 mg each by 30 min intravenous infusion 2 h before transplantation and day 4 after transplantation. The outcomes were compared with those of 8 children patients with leukemia who received haploidentical bone marrow transplantation without CD(25) antibody for GVHD prophylaxis.</p><p><b>RESULTS</b>All patients were engrafted and sustained full donor-type engraftment. 100% donors hematopoietic cells after transplantation was determined by cytogenetic evidence analysis. The median days of granulocyte exceeding 0.5 x 10(9)/L and pallets exceeding 20 x 10(9)/L were 19 and 22 days, respectively. Patients were monitored till up to days 100 for the sign of aGVHD. None developed the grade II-IV acute GVHD. However, the incidence of the grade II-IV acute GVHD in control group was 50% (P = 0.0147). Eight patients could be evaluated for chronic GVHD. All experienced chronic GVHD confined to the skin. The median follow-up duration was 12 months (range 9 - 24 months). Two patients died from transplant related mortality, one had patient relapsed disease and died. The remaining seven patients were alive in disease-free situation.</p><p><b>CONCLUSION</b>The use of Simulect in haploidentical bone marrow transplantation for the treatment of children patients with leukemia is effective for preventing acute severe GVHD and may reduce transplant-related mortality.</p>

Clinical study on hematopoietic reconstitution in patients with leukemia by haploidentical bone marrow transplantation / 中国实验血液学杂志

To investigate the properties of haploidentical donor-derived bone marrow engraftment and hematopoietic reconstitution in patients received bone marrow transplantation, 15 patients with leukemia received bone marrow grafts without T cell depletion from their family donors of those with 2-3 mismatched loci of HLA antigens. The donors were given G-CSF 250 micro g/day for 7 days prior to marrow harvest. All patients were treated with conditioning regimens consisting of high-dose of Ara-C, cyclophosphamide, and total body irradiation. A four-agent based GVHD prophylaxis was used as cyclosporine A, MTX, ATG and mycophenolate mofetile (MMF). Donor engraftment was evaluated as identification of HLA locus, chromosome karyotype, DNA fingerprinting, blood type and other parameters such as occurrence of GVHD, recovery of peripheral blood cell counts as well as normal myelogram. The results showed that successful and stable engraftment was established in all patients. The median time of granulocyte counts > 0.5 x 10(9)/L and platelet > 20 x 10(9)/L was 18 (13-23) and 22 (16-32) days, respectively. One of the patients relapsed despite the bone marrow chimerism appearing after transplantation. The grade I acute GVHD occurred in 8 and grade II-IV in 5 of the 15 patients. Of the patients, 7 received marrow grafts from donors of opposite sex were identified for donor engraftment by chromosome analysis, 4 by blood typing, 7 with HLA locus analysis and 1 with DNA fingerprinting. In conclusion, HLA haploidentical bone marrow transplantation is feasible with a series of management including mobilization with G-CSF in donors, intensive conditioning and proper immunosuppressants, which enable the allo-transplants to stride across the immunological barrier.

Clinical study of a new protocol for acute graft-versus host disease prophylaxis in HLA-haploidentical bone marrow transplantation (BMT) / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the feasibility of a new protocol for acute graft versus host disease (aGVHD) prophylaxis in haploidentical bone marrow transplantation.</p><p><b>METHODS</b>Thirty-eight high-risk leukemia patients underwent haploidentical G-CSF primed bone marrow transplantation without ex-vivo T cell depletion, the donors were given G-CSF 250 microg/d for 7 days prior to marrow harvest. All patients received a same chemo-radiation conditioning regimen, including cytarabin, cyclophosphamide and total body irradiation (TBI). GVHD prophylaxis regimen consisted of ATG, CsA, MTX, Mycophenolate mofetil (MMF) and anti-CD(25) monoclonal antibody (MoAb).</p><p><b>RESULTS</b>All patients achieved engraftment of a median of 19 and 22 days for neutrophil and platelet, respectively. Cytogenetic analysis showed 100% donor hematopoietic cells in all recipients after transplantation. One of the twenty patients (5%) experienced grades II - IV acute GVHD. The recovery of CD(3)(+)CD(8)(+) T cells, CD(19)(+) cells and CD(56)(+) cells after transplantation was within 3 approximately 12 months. Of the 20 patients, 16 were alive with minimal and limited chronic GVHD and karnofsky score over 90% in a median follow-up of 9 months. Disease free survival (DFS) rates was (80 +/- 9)%.</p><p><b>CONCLUSION</b>G-CSF priming marrow graft along with sequential immunosuppressants, especially the addition of anti-CD(25) MoAb for aGVHD prophylaxis could achieve excellent engraftment, proper immune reconstitution and very low incidence of grade II - IV GVHD. The new protocol is effective and feasible in preventing severe acute GVHD and improving DFS.</p>

Successful engraftment of T-cell undepleted haploidentical transplants by donor primed with G-CSF and additional use of ATG and MMF for recipients / 中国实验血液学杂志

Seventeen patients and their family donors HLA 2 - 3 antigen mismatched of 2 - 3 loci were enrolled in the study of haploidentical transplants from February 1999 to March 2001. Among patients with leukemia, most patients were classified as high risk. Eleven patients with ALL were all in more than second remission but one was in relapse. Patients with AML were one in CR1, one in CR2 while 4 patients with CML were two in CP and two in AP. The male-to-female ratio was 14:3 and the median age was 15 (range from 8 to 35). Conditioning regimens included Ara-C 3.0 g/m(2), 2 times per day x 3 d, on day 7, 6 and 5 pre-transplantation, CTX 45 mg/(kg per d) x 2 d on day 5 and 4 pretransplantation. TBI with 1000 cGy by 2 fractions on day 2 and 1 pretransplantation. The fresh and unmanipulated marrow was infused on day 0. Donors were received G-CSF (Lenograstim) at 3 - 4 microg/(kg per d) x 7 d. The BM cells were collected on eighth day. In GVHD prophylaxis, CSA, MTX, ATG (Antithymocyte globulin, Rabbit Fresenius S) and MMF (mycophenolate mofetic) were used in different periods. The dose of CSA was 1.5 mg/(kg per d) on day 7 to 1 pretransplantation, then 3 mg/(kg per d) from day 1 pretransplantation. MTX was 15 mg/m(2) on day 1 and 10 mg/m(2) on day 3, 6 and 11 posttransplantation. ATG was administered day 4 to 1 pretransplantation at 5 mg/(kg per d) and MMF dose was 1.0 g/d from day 7 posttransplantation. All patients established successful engraftment after initial transplantation. The median days of neutrophil exceeding 0.5 x 10(9)/L and platelet exceeding 20 x 10(9)/L were 18 (range 13 - 23) and 20 (range 16 - 32) days, respectively. Patients were monitored up to day 100 for the sign of aGVHD. The established grades II to IV aGVHD occurred in 5 out of 17 patients (29.4%). Eleven patients were surviving at a median follow-up of 13 months (range 3 - 27 months). Six out of the 17 patients died those 3 of them died of severe aGVHD, 2 of infection and 1 of leukemia relapse. Severe regimen-related toxicities were not experienced in all patients. The median follow-up period was 13 (range 3 - 27) months. Eleven patients were alive in disease-free situation with a Karnofsky performance status of 100%. This could be caused by the low overall incidence of aGVHD as a result of BM primed with G-CSF. The four-agent of immunosuppressive combined prophylaxis against GVHD in different periods may be highly effective.

A clinical study of haploidentical and G-CSF primed bone marrow transplantation by using CD25 for aGVHD prophylaxis / 中国实验血液学杂志

To explore the feasibility of using CD25 monoclonal antibody (Basiliximab) in T-cell undepleted allo-BMT with graft from haplotype-matched related donor for acute GVHD prophylaxis. Twenty-eight patients with leukemia received allo-BMT from HLA two or three loci mismatched related donors. The donors were given G-CSF (Lenograstim) 250 micro g/d for 7 doses prior to marrow harvest. CSA, MTX, ATG and mycophenolate mofetil (MMF) were combined for GVHD prophylaxis. ATG 5 mg/(kg.d) was infused from day 4 to day 1 before transplantation and MMF was administered from day 7. In the study group, the patients received additional CD25 monoclonal antibody for aGVHD prophylaxis. CD25 20 mg each by 30 min intravenous infusion on 2 hours before transplantation and day 4 after transplantation was administered while no application of CD25 in the controls. The outcomes of transplantation were compared between the stud y and control groups. The results showed that the median number of CD34(+) cell in graft was 5.9 x 10(6)/kg in the control group and 7.9 x 10(6)/kg in the study group. The median number of CD3(+) cell was 48 x 10(6)/kg and 52 x 10(6)/kg respectively (P > 0.05). All patients showed 100% donor-typed hematopoietic cells after transplantation by cytogenetic evidence. Five out of fifteen patients in the control group experienced II - IV acute GVHD. While none of thirteen in the study group developed the II - IV acute GVHD. However, none in both groups developed extensive cGVHD. The median follow-up duration was 8 (3-15) months in the study group and 26 (15-36) months in control. In the study group, one patient died from transplant related mortality (CMV infection); no one relapsed; and 12/13 patients survived in disease-free situation within the period of follow-up. In the control group, six patients died from transplant related mortality (3 GVHD, 2 infection and 1 relapsed) and 9/15 patients survived in disease-free situation. The one-year probabilities of disease-free survival (DFS) in two groups were significantly different (P < 0.05). It is concluded that the transplant from haploidentical donors used CD25 antibody is effective and feasible in preventing acute severe GVHD and improving DFS. The major histocompatibility barrier in the haploidentical related allo-BMT could be crossed by donors primed with G-CSF and GVHD prophylaxis with CD25 antibody in the hosts.

Allogeneic bone marrow transplantation for chronic myeloid leukemia using HLA identical sibling donors primed with G-CSF / 中国实验血液学杂志

Many studies have shown that G-CSF mobilized peripheral blood progenitor cell transplants (PBPCT) manifests faster recovery kinetics than conventional bone marrow transplants. This potential advantage of PBPCT still needs to be balanced against the risk of acute and chronic GVHD associating with the infusion of 10 - 15 fold higher donor lymphocyte number in unmanipulated allogeneic PBPCT than the marrow graft. To evaluate the effect of G-CSF primed bone marrow as a source of stem cells in the HLA-matched sibling transplantation, G-CSF primed with non-primed donor marrow in engraftment and incidence of GVHD for a homogenous group of patients with chronic myeloid leukemia (CML) were compared. Fifty patients with CML underwent bone marrow transplant, thirty-two donors (study group) were given G-CSF 3 - 4 micro g/kg per day for seven days prior to marrow harvest and eighteen donors (control group) had marrow harvest without G-CSF stimulation. Conditioning regimen consisted of total body irradiation and cyclophosphamide (CY), busulfan and CY, or busulfan, total body irradiation and CY. Both groups received same post-grafting GVHD prophylaxis and postgrafting G-CSF treatment. It was found that G-CSF primed donor marrow yielded with significantly higher number of total nucleated cells as well as CD34(+) cells and CFU-GM compared to non-G-CSF primed marrow (P = 0.001). The median engraftment time for absolute neutrophil (ANC > 0.5 x 10(9)/L) was day 15 (range 10 - 22) in the group of G-CSF primed vs day 21 in the non-primed donor group (P = 0.001). The median time for platelets (> 20 x 10(9)/L) was day 17.5 (range 13 - 28) in the group of G-CSF primed vs day 24 in non-primed group (P = 0.001). The incidence of acute GVHD grade II - IV in G-CSF primed donor group was surprisingly as low,as only two cases of thirty-two transplants (6.3%) with acute GVHD grade II limited to the skin. Whereas, five of eighteen patients (27.8%) in the control group developed acute GVHD grade II - IV (P = 0.032). G-CSF primed donors showed reduced CD4(+) and increased CD8(+) cells, resulting in a significant reduction of CD4(+)/CD8(+) ratio as compared with non-primed marrow. The total CD3(+) cell count kept unchanged in G-CSF primed donors. There were not significant differences in the incidence of the chronic GVHD (24% vs 33.3%), relapse rate (12.5% vs 11.1%) and overall survival rate (78.1% vs 66.7%, P = 0.32) during 6 - 50 months of follow-up. In conclusion, G-CSF primed donor marrow accelerates engraftment. Although G-CSF did not change the total CD3(+) cells in bone marrow, it altered the ratio of CD4(+) and CD8(+) cells and significantly reduced the incidence of acute GVHD.

Impact on IL-2 expression of lymphocytes in donors after G-CSF administration and its clinical significance / 中国实验血液学杂志

In this study, to investigate the effect on expression of IL-2 in lymphocytes from bone marrow and peripheral blood of normal donors after they were mobilized by G-CSF in allo-BMT, 7 normal donors bone marrow and peripheral blood were harvested before and after G-CSF administration. The separated lymphocytes were measured by FCM after they were stained intracellularly by anti-IL-2, and their expressions of IL-2 were compared. The degree of aGVHD in patients after bone marrow transplantation was evaluated clinically, and it was compared with the status of aGVHD of 15 patients whose donors didn't receive G-CSF administration in our department, and 2 groups of patients are comparable in age, types of diseases and status of donors. The results showed that the expression of IL-2 in lymphocytes in 7 G-CSF mobilized donors decreased significantly after G-CSF administration and more severe aGVHD than grade II didn't develop in these recipient patients, and comparing with 15 patients received the bone marrow from donors who didn't receive G-CSF, the incidence of aGVHD decreased. It is suggested that the expression of IL-2 in lymphocytes was influenced by donors' G-CSF administration, and it is likely that thereby reduces the incidence of aGVHD in patients after BMT.

Correlation of bone marrow stromal function in vitro with efficacy of anisodamine therapy for aplastic anemia / 中国实验血液学杂志

To observe normal bone marrow stromal function for supporting hematopoiesis and realize the correlation of bone marrow stromal function in patients with aplastic anemia (AA) and anisodamine therapy, normal human marrow CFU-GM was assayed on bone marrow stromal layer (SL) formed on week 3 culture, and the CFU-GM without SL as a control (100%). Results showed that the production of CFU-GM on the normal SL was significantly higher than that of the control. The production of CFU-GM on the SL of 4 AA patients, who responded to anisodamine, was < 80% of the control, and when 2 of them were reexamined for stromal function after treatment, the number of CFU-GM on SL rose up to 168.8% and 249.2% from 38.7% and 39.7% of the control respectively. While in the rest 6 patients, the number of CFU-GM was > 80% of the control, only one patient was improved by anisodamine therapy. So, normal bone marrow stromal layer can obviously support the growth of granulocyte/macrophage progenitor cells. Anisodamine therapy could be an effective agents for aplastic anemia with stromal dysfunction

Short Course High Dose Mitoxantrone with Cytarabine for Treatment of Refractory and Relapsed Acute Leukemia / 中国实验血液学杂志

Although chemotherapy can achieve a high rate of disease remission in patients with newly diagnosed acute leukemia, patients with recurrent or refractory leukemia generally have a poorer rate of response. This study was designed to assess the utility of high-dose mitoxantrone and intermediate-dose cytarabine in the treatment of patients with relapsed or refractory acute leukemia. Thirty patients with relapsed or refractory acute leukemia were treated with mitoxantrone at a total dose of 40 mg/m(2) intravenously and cytarabine 1 - 1.5 g/m(2) over 3 hours once daily for five doses. Twenty-six of thirty patients achieved complete response (CR 87%) and one achieved partial response (PR 3.3%). No patient died during the induction course. The main toxic effect was hematopoietic suppression that was clinically acceptable. The median time needed for CR was around 30 days. The median disease-free and overall survival times were 3.5 and 6 months, respectively. The results demonstrate that short course high-dose mitoxantrone with cytarabine is associated with a trend toward a higher CR rate and therefore, it should be an effective antileukemic regimen for the treatment of refractory and relapsed acute leukemia.