ABSTRACT
AIM: To investigate the role of transcription factor c-Ets1 in cyclin D3 expression and its effects on the proliferation of umbilical cord hematopoietic cells. METHODS: Cyclin D3 promoter deletion constructs were generated and transfected into CD34(+) cells. Dual luciferase reporter assays and TFSEARCH software were used to identify negative regulatory domains and to predict putative transcription factors involved in cyclin D3 downregulation. Expression of c-Ets1 in CD34(+) cells was detected using electrophoretic mobility shift and super shift assays. Point mutants of c-Ets1 binding sites were constructed. The wild-type c-Ets1 and the mutant promoter constructs were co-transfected into CD34(+) cells to determine the promoter activity. The impact of c-Ets1 expression on the proliferation of CD34(+) cells was assessed using MTT assay. RESULTS: Nine cyclin D3 promoter deletion constructs were generated. A negative regulatory domain containing c-Ets1 binding sites was identified between -439 bp and -362 bp. Transfection of the promoter deletion constructs containing mutant c-Ets1 binding sites enhanced cyclin D3 promoter activity. However, the opposite results were observed when CD34(+) cells were co-transfected with wildtype c-Ets1 and its promoter deletion constructs. The overexpression of c-Ets1 could suppress cyclin D3 mRNA and protein levels. In addition, it inhibits the proliferation of CD34(+) cells. CONCLUSION: c-Ets1 functions as a negative transcription factor, down-regulating the expression of cyclin D3, which leads to inhibition of CD34(+) cell proliferation.
Subject(s)
Cyclin D3/genetics , Fetal Blood/cytology , Gene Expression Regulation , Hematopoietic Stem Cells/cytology , Proto-Oncogene Protein c-ets-1/metabolism , Binding Sites , CD3 Complex/analysis , Cell Proliferation , Cells, Cultured , Humans , Point Mutation , Proto-Oncogene Protein c-ets-1/geneticsABSTRACT
Haploidentical hematopoietic cell transplantation (HCT) has been used to treat hematologic malignancies, but it is unknown whether the procedure is more effective in adults or children. To address this question, we analyzed patients aged 1 to 65 years old receiving myeloablative conditioning regimens followed by family 2 to 3 antigen HLA-mismatched HCT and reported to the Center for International Blood and Marrow Transplant Research (CIBMTR; n = 137) or performed in Dao-Pei Hospital in China, China (n = 181). The Dao-Pei cohort had more acute and chronic graft-versus-host disease (GVHD), less relapse, lower transplant-related mortality (TRM), and better leukemia-free survival (LFS) than the CIBMTR cohort. Overall survival (OS) and outcomes were similar between adults and children. In the CIBMTR cohort receiving ex vivo T cell depletion (TCD), adults had higher TRM (relative risk [RR] 2.71, 95% confidence interval [CI] 1.29-5.69, P = .008) and lower OS (RR 1.75, 95% CI 1.08-2.84, P = .023) than children. In the CIBMTR subset that did not receive ex vivo TCD, relapse was lower in adults compared to children (RR 0.24, 95% CI 0.07-0.80, P = .020), but TRM, LFS, and OS were similar. We conclude that outcomes in adults and children are similar overall, although children have better survival than adults if ex vivo TCD is used.
Subject(s)
Hematologic Neoplasms/surgery , Hematopoietic Stem Cell Transplantation/methods , Adolescent , Adult , Age Factors , Aged , Child , Child, Preschool , Cohort Studies , Disease-Free Survival , Female , Haplotypes , Hematologic Neoplasms/genetics , Hematologic Neoplasms/immunology , Humans , Infant , Male , Middle Aged , Multivariate Analysis , Survival Rate , Transplantation Conditioning/methods , Treatment OutcomeABSTRACT
SUMMARY: This report describes a safe and effective therapy through adoptive transfer of donor cytomegalovirus (CMV)/Epstein-Barr virus (EBV) immune effector cells. The patients, from 3 to 10 years of age, suffering from hematologic diseases received haploidentical transplantation. All 3 patients developed varying levels of viremia from days 13 to 31 and 2 patients developed CMV-interstitial pneumonitis or interstitial inflammation after transplantation. Tapering down the dose of immunosuppressives together with intensive antivirus therapy and escalated infusions of donor-derived CMV/EBV immune effector cells effectively controlled virus-related diseases. All 3 patients survived and remained CMV/EBV-free 14-16 months after transplantation.
Subject(s)
Adoptive Transfer/methods , Cytomegalovirus/immunology , Herpesvirus 4, Human/immunology , Premedication/methods , Child , Child, Preschool , Cytomegalovirus Infections/etiology , Cytomegalovirus Infections/prevention & control , Dendritic Cells/transplantation , Dendritic Cells/virology , Epstein-Barr Virus Infections/etiology , Epstein-Barr Virus Infections/prevention & control , Female , Hematologic Neoplasms/complications , Hematologic Neoplasms/therapy , Hematopoietic Stem Cell Transplantation/adverse effects , Humans , Male , Survival Rate , T-Lymphocytes, Cytotoxic/transplantation , T-Lymphocytes, Cytotoxic/virology , Treatment OutcomeABSTRACT
The objective of this study was to investigate the effect of ligustrazine on the expression of stem cell factor mRNA (SCF) in bone marrow tissue and explore the mechanism of hematopoietic reconstitution after bone marrow transplantation (BMT). The colony forming unit of spleen (CFU-S) were counted, the survival rate at days 7, 14 and 21 after BMT were measured, as well as the expression level of SCF mRNA was detected by RT-PCR. The results showed that in ligustrazine group CFU-S counts on day 10 and survival rate, expression level of SCF mRNA on day 7, 14 and 21 after BMT were higher than that in the control group (p < 0.01 or p < 0.05). In conclusion, ligustrazine promotes the recovery of hematopoietic cells in bone marrow, enhances the repair of bone marrow microvessels, and then improves bone marrow microenvironment and promotes hematopoietic reconstitution.
Subject(s)
Bone Marrow Transplantation , Hematopoiesis/drug effects , Pyrazines/pharmacology , Animals , Male , Mice , Mice, Inbred BALB C , RNA, Messenger/genetics , RNA, Messenger/metabolism , Stem Cell Factor/genetics , Stem Cell Factor/metabolism , Transplantation, IsogeneicABSTRACT
To explore the hematopoiesis inhibition mechanisms of interferon-gamma (IFN-gamma), the effects of IFN-gamma on the expression of the cyclin D in the umbilical cord blood hematopoietic stem/progenitor cells were observed. In the experiments the CD34(+) cells were isolated from the cord blood with MIDI-MACS system; semi-solid methylcellulose culture technique was used to measure the formation of CFU-GM; the expression levels of cyclin D isoforms were assayed by semi-quantitative RT-PCR, after the hematopoietic stem/progenitor cells were incubated with IFN-gamma. The results indicated that IFN-gamma could inhibit the formation of CFU-GM and down-regulate the expression of cyclin D2 and cyclin D3 at the mRNA level. It is concluded that the IFN-gamma could inhibit the proliferation of hematopoietic stem cells and down-regulate the expression of cyclin D, that may be one mechanism underlying the hematopoietic inhibition of IFN-gamma.
