OBJECTIVETo analyze the risk factors of patients with relapsed leukemia after allogeneic hematopoietic stem cell transplantation, and to explore the therapeutic strategies for recurrence.

METHODSThe Cox proportional hazard regression model was used for univariate and multivariate analysis of transplantation-related index, a single center retrospective study of clinical data of 202 cases of leukemia received allo-HSCT from March 2004 to October 2014 had been conducted to screen the risk factors for recurrence after transplantation.

RESULTSIn the leukemia patients received allo-HSCT, 68 cases relapsed. The relapse rate was 33.6%. The median time of relapse was 4(1.5-26 ) months. Univariate analysis indicated that there were 5 risk factors related with the disease relapse(P<0.05), including the type of disease, extramedullary disease prior to transplant, the course of induced remission, the status of disease at HSCT and chronic graft versus host disease(cGVHD). Multivariate analysis showed that extramedullary disease prior to transplant(RR=2.622, 95%CI 1.139-6.037), the course of induced remission(RR=1.156, 95%CI 0.682-1.957), cGVHD (RR=1.728,95%CI 0.999-2.991) were independent risk factors for relapse of the patients received transplantation. Treatment strategies for the relapsed patients included withdraw immunosuppressant, donor lymphocyte infusion, systemic chemotherapy and local radiotherapy, targeted therapy, and second transplantation. Individualized choice was needed according to the relapsed site. The relapse-related mortality was 25.2%.

CONCLUSIONThe relapsed patients with leukemia after allo-HSCT have poor prognosis, early interference has good effect. The evaluation and prevention of risk factors before transplantation is even more important.

Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunosuppressive Agents ; Leukemia ; Multivariate Analysis ; Proportional Hazards Models ; Recurrence ; Remission Induction ; Retrospective Studies ; Risk Factors ; Transplantation, Homologous

OBJECTIVETo explore the safety and efficiency of unrelated donor peripheral blood stem cells (URD-PBSC) transplantation combined with umbilical cord mesenchymal stem cells (UC-MSC).

METHODSThe clinical data of 49 patients received unrelated donor peripheral blood stem cells transplantation (URD-PBSCT) for treating hematologic malignancies were retrospectively evaluated, including 12 ANLL, 17 ALL, 18 CML and 2 MDS. Out of them, 22 patients received the URD-PBSCT combined with UC-MSC and 27 patients received only URD-PBSCT. The average number of infusing UC-MSC was 1.0 × 10⁶/kg in the UC-MSC+URD-PBSCT group.

RESULTSAs compared with URD-PBSCT group, in UC-MSC+URD-PBSCT group the median recovery time of neutrophilc granulocytes was shorter (12 d vs 15 d) (P = 0.041), the incidence and severity of chronic graft versus host disease (cGVHD) were lower (20.0% vs 51.9%) (P = 0.026) (5.0% vs 33.3%) (P = 0.040), the incidence of CMV infection after transplantation was higher (81.8% vs 51.9%) (P = 0.028). In addition to these, the differences were not statistically significant in term of implantation level, PLT reconstitution, aGVHD, lung infection, hemorrhagic cystitis, 1-year relapse and survival between the 2 groups (P > 0.05).

CONCLUSIONThe transplantation of URD-PBSC combined with UC-MSC is effective and safe. The speed of neutrophils reconstitution is faster. The incidence and severity of cGVHD are lower, but the attention should be paid to prevent the CMV infection.

Cytomegalovirus Infections ; Graft vs Host Disease ; Hematologic Neoplasms ; therapy ; Humans ; Incidence ; Mesenchymal Stem Cell Transplantation ; Neoplasm Recurrence, Local ; Peripheral Blood Stem Cell Transplantation ; Retrospective Studies ; Umbilical Cord ; cytology ; Unrelated Donors

This study was purposed to explore the therapeutic efficacy and influencing factors of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with chronic myelomonocytic leukemia (CMML) and in patients with juvenile myelomonocytic leukemia (JMML). The clinical data of 3 cases of CMML and 2 cases of JMML underwent allo-HSCT were analysed in term of multiparameter. The results showed that the hematopoietic stem cells in 5 patients grafted successfully. One case of JMML died of pulmonary disease, other 4 cases survive without disease. The analysis found that the disease burden before transplant, chromosome karyotype, acute GVHD II-IV and poor risk cytogenetics all associated with the relapse rate and disease-free survival rate of CMML. The low intensity conditioning regimen was better than myeloablative conditioning regimen. Type of donor and source of stem cells did not statistically and significantly affect OS and RFS. The splenectomy before allo-HSCT as well as spleen size at time of the alloHSCT did not influence on posttransplantation outcome of JMML. However, cord blood HSCT for JMML patients delayed hematologic recovery as compared to that of bone marrow or peripheral blood HSCT. The age, GVHD, HbF level played an important role in leukemia replace. It is concluded that the allogeneic hematopoietic stem cell transplantation is a curative regimen for CMML and JMML, but there also is a serial problems to be resolved.
Adult ; Child, Preschool ; Hematopoietic Stem Cell Transplantation ; Humans ; Infant ; Leukemia, Myelomonocytic, Chronic ; therapy ; Leukemia, Myelomonocytic, Juvenile ; therapy ; Male ; Middle Aged ; Transplantation, Homologous ; Treatment Outcome

This study was aimed to compare the differential expressions of calcineurin (PP2B, PP3) in the mouse Pre-B cell lines (S9) and the tumor cell lines (S4C2) derived from pre-B lymphocytes, and to clarify its possible mechanism involving in the leukemia cell apoptosis. The quantitative real-time PCR was used to detect the differential expressions of H2AX-associated phosphakinase ATM, ATR, DNA-PKs, JNK1, P38 and the γ-H2AX-related phosphatase PP1, PP2A, calcineurin, PP4, PP6, PP5 between S9 and S4C2 cell lines. CCK-8 assay and flow cytometry were used to detect the effect of imatinib (IM) and cyclosporine A (CsA) on cytotoxicity and apoptosis of 2 cell lines. The Western blot was used to detect the effects of 2 drugs on apoptosis of S9 and S4C2 cell lines. The results showed that the expression level of calcineurin gene in the leukemia cell S4C2 was about 3.5 times of that in S9 cells, while the expression of other genes in these 2 kinds of cells was not significantly different. The apoptosis and toxicity of IM and CsA on S4C2 cells was significantly stronger than that on S9 cells. The expression level of calcineurin in S4C2 cells was higher than that in S9 cells.When CsA inhibited the calcineurin activity, the expression of DNA damage marker γ-H2AX in S9 cells was significantly lower than that in S4C2 cells,while the expression level of γ-H2AX between the two cell lines was no significantly different after treatment with imatinib, the expression level of γ-H2AX in S9 cells was lower than that in S4C2 cells when the two drugs were combined. It is concluded that the calcineurin plays a role of anti-apoptosis in B leukemic cells, cyclosporine A can promote the leukemia cell apoptosis.
Animals ; Apoptosis ; Calcineurin ; metabolism ; Cell Line, Tumor ; Cyclosporine ; DNA Damage ; Flow Cytometry ; Leukemia ; metabolism ; Mice ; Precursor Cells, B-Lymphoid ; metabolism ; Real-Time Polymerase Chain Reaction

This study was purposed to investigate the therapeutic effects of early transfusion of immunized donor lymphocytes after haploidentical transplantation by means of mouse model of nonmyeloablative haploidentical bone marrow transplantation. CB6F1 female mouse was served as recipient and C57BL/6 male mouse was served as donor. Each CB6F1 female mouse was subjected to intravenous transfusion with 1×10(6) erythroleukemia (EL9611) cells at day 4 before transplantation, followed with intraperitoneal injection of Ara-C (0.015 g) respectively at day 2 and day 1, then conditioned for BMT with TBI (450 cGy) at day 1 before transplantation. After conditioning (day 0), each of recipients was transplanted with 6×10(7) mixture of bone marrow and spleen cells from the C57BL/6 mice, and was infused with 6 × 10(7) immunized donor lymphocytes at day 15 after transplantation. All treated animals were evaluated for survival, development of leukemia and aGVHD. The donor CD3(+) cell chimerism and sex determining region Y gene (SRY)in recipients were monitored periodically after transplantation. The results showed tht all mice with only inoculation of 10(6) EL9611 cells survived for 15 ± 1 days (n = 4); all mice of other groups obtained the varying degrees of implantation. SRY could be detected at day 30 and 60 after transplantation. The chimerism of donor CD3(+) cells in mixed bone marrow transplantation (MT) group at day 14, 30 and 60 respectively reached 17.95% ± 12.03%, 37.34% ± 2.78% and 47.06% ± 6.1%. In donor lymphocyte infusion (DLI) group it reached 69.78% ± 12.62%, 75% ± 15.97%, 83.41% ± 16.07% at day 30, 45 and 60 after transplantation. The mice of MT and DLI group survived for 66.66 ± 1.47 days and 78.2 ± 7.82 days. It is concluded that the high tumor burden before transplantation can affect donor cell engraftment and prognosis.Early post-transplanted infusion of immunized lymphocytes from donor can help to improve the therapeutic efficacy and survival.
Animals ; Bone Marrow Transplantation ; methods ; Female ; Haplotypes ; Leukemia, Erythroblastic, Acute ; therapy ; Lymphocyte Transfusion ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Tissue Donors ; Transplantation Conditioning ; methods ; Transplantation, Homologous

OBJECTIVETo detect the expression profile of NK ligands in acute leukemia cell lines and investigate the differential expression pattern between acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML).

METHODSUsing quantitative real-time PCR, 23 NK ligands (MICA, MICB, ULBP-1, ULBP-2, ULBP-3, ULBP-4, HLA-E, HLA-G, CD48, NBTA, HLA-F, LLT-1, PVR, Nectin2, CD72, CD80, ICAM-1, LFA-3, CRACC, Fas, DR4, DR5, TNFR1) were detected in 6 acute leukemia cell lines, including 3 ALL cell lines (CEM, Jurkat T, Reh) and 3 AML cell lines (HL-60, KG-1a, NB4), respectively. Independent-samples t test analysis was performed to determine statistical significance.

RESULTSUsing β-actin as reference gene, the relative expression results showed that the expression of 4 NK ligands between ALL and AML is significantly different. Specifically, the level of ULBP-2 is higher in ALL (CEM: 1, Jurkat T: 0.617, Reh: 0.246) than that in AML (HL-60: 0.000, KG-1a: 0.003, NB4: 0.000)(P = 0.047). However, the expressions of CD48, PVR(PVR-1, PVR-2) and DR4 is higher in AML (HL-60: 13.987, 4.403, 10.334, 8.711; KG-1a: 5.387, 2.900, 7.315, 4.512; NB4: 7.763, 3.248, 7.049, 6.127) than that in ALL (CEM: 1, 1, 1, 1; Jurkat T: 2.035, 1.553, 3.888, 0.449; Reh: 1.559, 0.000, 0.000, 1.304) (P = 0.044, 0.014, 0.014, 0.011). And there're no significant differences between the rest 19 NK ligands.

CONCLUSIONSULBP-2, CD48, PVR and DR4 might play an important role in the distinct mechanisms in leukemogenesis between ALL and AML and could be potential targets for diagnosis and treatment.

Acute Disease ; Antigens, CD ; genetics ; metabolism ; CD48 Antigen ; Cell Line, Tumor ; GPI-Linked Proteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Leukemia ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Ligands ; Membrane Proteins ; genetics ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; metabolism ; Receptors, Virus ; genetics ; metabolism

Bone Marrow Transplantation ; immunology ; Case-Control Studies ; Haploidy ; Humans ; Immunity, Cellular ; immunology ; Recovery of Function ; Transplantation Conditioning ; Transplantation, Homologous

This study was aimed to explore the difference of NK cell receptor NKG2D and NKG2A expression on NK cells and CD3(+) T cells and their ligand MHC-I A/B (major histocompatibility complex class I-related chains A/B) and HLA-E expression in leukemia cells, as well as its immunological significance. Flow cytometry was used to detect the killing rate of NK92 cells to 8 leukemia cell lines, and the expression of NKG2D and NKG2A on NK cells and CD3(+) T cells as well as their ligand MHC-I A/B and HLA-E expression on leukemia cells. The results indicated that the NK92 showed different killing activity to different leukemia cell lines. The positive expression rate of NKG2D and NKG2A on NK cells and CD3(+) T cells in ALL patients was no significantly different from that in AML patients (p > 0.05), but positive expression rate of MHC-I A/B and HLA-E in ALL patients was obviously higher than that in AML patients (p < 0.05). It is concluded that there is difference of immune cell function between ALL and AML patients, this difference may be associated with the expression difference of NKG2D and NKG2A ligands on leukemia cells while does not associated with the killing and inhibiting receptors expressed on NK cells and CD3(+) T cells.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Histocompatibility Antigens Class I ; genetics ; metabolism ; Humans ; Infant ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male ; Middle Aged ; NK Cell Lectin-Like Receptor Subfamily C ; genetics ; metabolism ; NK Cell Lectin-Like Receptor Subfamily K ; genetics ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Young Adult

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.
Hematopoietic Stem Cell Transplantation ; Humans ; Immune Tolerance ; T-Lymphocytes, Cytotoxic

The aim of this study was to investigate the efficacy and feasibility of anti-CD25 monoclonal antibody (basiliximab) in treating steroid-refractory acute graft-versus-host disease (aGVHD) following haploidentical bone marrow transplantation (hiBMT). 15 cases who developed II-IV grade steroid-resistant aGVHD after haploidentical BMT were treated by intravenous injection of basiliximab at a dose of 20 mg on days 1 and 4. In those patients not achieving CR after 1 week, basiliximab injection was repeated. The results showed that 8 cases (53.33%) got complete response (CR). Out of them 4 cases have been still in disease-free survival, 2 cases have been in survival with limited cGVHD, 2 cases died from pulmonary infection; 3 cases (65%) got partial response (PR), out of whom 1 case has been still in disease-free survival, one died from GVHD and infection, and another one died from pulmonary infection; 4 cases without response died from GVHD, pulmonary infection and cardiac failure. Overall response rate was 73.3% and long-term survival rate was 46. 7%. There were no infusion-associated side-effects after treatment with basiliximab. It is concluded that the anti-CD25 monoclonal antibody is efficacious and feasible for steroid-refractory grade II-IV aGVHD after hiBMT, but the overall survival rate is low. Infection is the main cause of death. Thereby, it is especially important to strengthen environmental protection and prevent infection.
Adolescent ; Adult ; Antibodies, Monoclonal ; therapeutic use ; Bone Marrow Transplantation ; adverse effects ; Child ; Child, Preschool ; Drug Resistance ; Female ; Graft vs Host Disease ; drug therapy ; Hormones ; pharmacology ; Humans ; Interleukin-2 Receptor alpha Subunit ; immunology ; Recombinant Fusion Proteins ; therapeutic use ; Treatment Outcome ; Young Adult