To clarify the hematopoietic potential of various sub-classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with anti-CD34 and -CD38 McAb. We characterized two cell fra...To clarify the hematopoietic potential of various sub-classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with anti-CD34 and -CD38 McAb. We characterized two cell fractions in CD34+cells with or without CD38 expression. A clonogenic assay showed that most CFC were present in CD34+CD38+ population. Morphologic analysis showed that blast-like cells were more enriched in the CD34+CD38 fraction. To clarify the biologic differences between both fractions, we examined the more primitive progenitor cell function by assessing long-term culture-initiating cells (LTC-IC) on the stromal cells. At the first two weeks, more CF.C harvested from the culture in the fractions initiated with both populations. However, more LTC-IC were present during weeks 4 to 12 in the CD34+CD38- population. These results indicate the primitive progenitors are more enriched in CD34+CD38 population than in CD34+CD38+ cells.展开更多
Antitumor chemotherapy is often limited by hematopoietic toxicity.TO attenuate themyelosuppressive effects of chemotherapy, successfulengraftment of unmodified fetal liver cells (FLCs) hasbeen achieved across allogene...Antitumor chemotherapy is often limited by hematopoietic toxicity.TO attenuate themyelosuppressive effects of chemotherapy, successfulengraftment of unmodified fetal liver cells (FLCs) hasbeen achieved across allogeneic barriers in a number ofanimal models and patients, GM-CSF is a展开更多
Previous reports indicated that the difference ofMHC-Ⅰ and Ⅱ between donor and recipient was criticalfor the occurrence of allogenic rejection or graft versushost reaction (GVHR). This study aimed at looking forthe ...Previous reports indicated that the difference ofMHC-Ⅰ and Ⅱ between donor and recipient was criticalfor the occurrence of allogenic rejection or graft versushost reaction (GVHR). This study aimed at looking forthe possibility of reducing the risk of immunologicalrejection through transplantation tolerance induced bytransduction of MHC gene. In the present study, wetransferred donor mice MHC class I gene (H-2k^b gene)展开更多
Stem cell factor (SCF) is a novel growth factor thatinfluences the growth and development of hematopoieticcells, germ cells and melanocytes. To explore
Liposomes have several advantages over viral vectorsfor gene delivery both in vitro and in vivo. However,few data are available concerning gene transfer intohematopoietic stem cells. In order to explore theefficiency ...Liposomes have several advantages over viral vectorsfor gene delivery both in vitro and in vivo. However,few data are available concerning gene transfer intohematopoietic stem cells. In order to explore theefficiency and the stability of expression of gene transferinto hematopoietic stem cells, we have transduced twomarker genes (Neo^R and Lac Z) co-transfer into humanbone marrow CD34^+ hematopoietic stem cells展开更多
The effect of recombinant human interleukin-6 ( rhIL-6) on the early phase of post-irradiation hematopoietic recovery ( mainly CFU-S, CFU-GM and CFU-E) has been studies in 6.5 Gy gamma-irradiated C57BL / 6J mice, whic...The effect of recombinant human interleukin-6 ( rhIL-6) on the early phase of post-irradiation hematopoietic recovery ( mainly CFU-S, CFU-GM and CFU-E) has been studies in 6.5 Gy gamma-irradiated C57BL / 6J mice, which had been given subcutaneous injection of rhIL-6 b. i. d. at 10, 100, 200, 500 and 1 000μg/kg/d for 4 or 6 consecutive days started from 30 min after irradiation. It was shown that the effect of IL-6 on the early phase of post-irradiation hematopoietic recovery is dose-dependent and diphasic. At the doses of 200-1000μ/kg/d, IL-6 had a favorable effect on post-irradiation hematopoietic recovery expressed as increse of spleen weight and CFU-S, and elevation of CFU-GM yield and total CFU-GM content in femur. Administration of relatively low doses of IL-6(10-100 μg/kg/d) suppressed postirradiation hematopoietic regeneration. High a dose (1 000μg/kg/d) did not show any stronger stimulative effect on post-irradiation hematopoietic reconstitution. There is an optimal range of IL-6 dose for展开更多
Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemo-lytic disease in which there is a stem cell disorder of clonal nature. Previous studies have demonstrated that the numbers of burst-forming units-erythroid...Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemo-lytic disease in which there is a stem cell disorder of clonal nature. Previous studies have demonstrated that the numbers of burst-forming units-erythroid (BFU-E) and colony-forming units-granulocyte / macrophage (CFU-GM) from bone marrow of PNH patients growing in the medium containing PHA-LCM from the normai donors were more reduced than those of normai bone marrow. The purpose of present study was to investigate if PNH lymphocytes are defective in supporting hematopoiesis in vitro. PHA-LCM from PNH blood was added to the culture medium for the growth of PNH and normai BFU-E and CFU-GM. The numbers of PNH bone marrow BFU-E and CFU-GM in the medium containing PHA-LCM from PNH blood were less than those from normai blood; the numbers of normai bone marrow BFU-E and CFU-GM grown in the medium containing PHA-LCM from PNH blood were more decreased than those from normai blood. The results suggest that diminished numbers of PNH bone marrovv BFU-展开更多
In order to study the epidemiological characteristics of cytomegalovirus(CMV)infection in allogeneichematopoietic stem cell transplantation(allo-HSCT)recipients by means of plasma real time quantitative polymerasechai...In order to study the epidemiological characteristics of cytomegalovirus(CMV)infection in allogeneichematopoietic stem cell transplantation(allo-HSCT)recipients by means of plasma real time quantitative polymerasechain reaction(RQ-PCR),141 adult patients undergoing allo-HSCT between January 2008 and June 2010 were seriallymonitored by RQ-PCR for detecting CMV and guiding the preemptive therapy followed up to 180 days post-HSCT.Theresults showed that the incidence of CMV infection and CMV pneumonia was 81.5%and 2.9%respectively,whichmainly occurred within 2 months post-HSCT.Single-therapy with ganciclovir(GCV)for 63 patients or foscarnet 6patients was performed for preemptive therapy.The total efficacy was 87.8%,and the response patterns were different.CMV infection was more frequent in female patients(P=0.044),and those with aGVHD(P=0.043),using ATG orbasiliximab in conditioning regimens(P=0.049),as well as earlier in patients using ATG or basiliximab or those withaGVHD(P=0.007;P=0.000).The aGVHD,maximum load,positive times of CMV-DNA detection and therapyduration all correlated with the efficacy(P<0.05).It is concluded that the incidence of CMV infection is still high afterHSCT.Plasma RQ-PCR assay for CMV-DNA shows a strong correlation with the clinical outcome of CMV infection,which is useful and suitable for management of CMV infection in HSCT.展开更多
To elucidate the feasibility of retroviral mediatedtransfer of multidrug resistance gene (mdrl) intohematopoietic stem/progenitor cells, the CD34^+ cellswere isolated from bone marrow by using a high-gradient.magnetic...To elucidate the feasibility of retroviral mediatedtransfer of multidrug resistance gene (mdrl) intohematopoietic stem/progenitor cells, the CD34^+ cellswere isolated from bone marrow by using a high-gradient.magnetic cell sorting system, and the efficiency ofretroviral mediated gene transfer was studied.展开更多
It has been demonstrated that tbe critical role ofbone marrow stromal cells (HMSCs ) is to sustain theselfrenewal of pluripotent hematopoietic stem cells andmaintain the homeostasis of bone marrow hematopoiesismicroen...It has been demonstrated that tbe critical role ofbone marrow stromal cells (HMSCs ) is to sustain theselfrenewal of pluripotent hematopoietic stem cells andmaintain the homeostasis of bone marrow hematopoiesismicroenvironment. BMSC progenitor can differentiateinto several clements including macrophages, endothelialcells, fibroblasts and some other cells. Almost展开更多
Cancer gene therapy over the past several yearshas involved the introduction of genes intohematopoietic cells for: (1) protecting the normalcells from the side effects of chemotherapy; (2)introduction of genes into th...Cancer gene therapy over the past several yearshas involved the introduction of genes intohematopoietic cells for: (1) protecting the normalcells from the side effects of chemotherapy; (2)introduction of genes into the neoplastic cells for展开更多
Retroviral mediated gene transfer of humanglobin gene into hematopoietic stem cells is apromising approach for thalassemia gene therapy.Major problem of the transferred globin gene was lowlevel expression of the gene ...Retroviral mediated gene transfer of humanglobin gene into hematopoietic stem cells is apromising approach for thalassemia gene therapy.Major problem of the transferred globin gene was lowlevel expression of the gene with its proximal cis-acting sequence. The locus control region (LCR) ofthe human β-globin gene cluster consists of four majorDNase I hypersensitive sites (HS). When linked展开更多
Chemotherapy is one of the curative treatmentmodality for several types of tumors. However, thedosage of antitumor drugs was limited by cytotoxicityof drugs to normal tissues, especially for bone marrow(myelosuppressi...Chemotherapy is one of the curative treatmentmodality for several types of tumors. However, thedosage of antitumor drugs was limited by cytotoxicityof drugs to normal tissues, especially for bone marrow(myelosuppression). Drug resistantce gene (hMDRI,hDHFR etc.) is expressed in many normal tissues. Itsover-expression in tumor cells give rise to the tumorresistant to many drugs. But, if we transfer the drugresistance gene into the normal hematopoietic stemcell (HSC), the expression of the foreign gene展开更多
In order to increase the expression of mdrl genetransfectcd using retroviral vcctors, we constructedtwo retroviral vectors (LmdrlSN and MSCV-mdrl)containing mdrl cDNA and compared the expressionof mdrl gene with anoth...In order to increase the expression of mdrl genetransfectcd using retroviral vcctors, we constructedtwo retroviral vectors (LmdrlSN and MSCV-mdrl)containing mdrl cDNA and compared the expressionof mdrl gene with another vector pHamdrl/A(Pastan et al. Proc Natl Acad Sci USA 85: 4486,1986) in NIH.3T3 cells. The LmdrlSN was made展开更多
An extract (G-INH) made from mature human granulocytes freshly isolated from normai blood causes human neutrophils to undergo apoptosis in vitro as shown by morphologic changes and by the typical ladder pattern of sma...An extract (G-INH) made from mature human granulocytes freshly isolated from normai blood causes human neutrophils to undergo apoptosis in vitro as shown by morphologic changes and by the typical ladder pattern of small DNA fragments noted on agarose gel electrophoresis of isolated DNA. Apoptosis occurs in from 20% to 30% of neutrophils over 24 hours of culture in vitro and the addition of G-INH to the medium causes a dose-related increase in the incidence of apoptosis. Heating G-INH at 60t for 30 minutes does not destroy its capacity to induce apoptosis but GM-CSF, G-CSF, and to a lesser extent IL-1β, antagonize this action. IL-3 does not diminish G-INH induced apoptosis of neutrophils. Substances, released from, mature neutrophils may participate in regulating the survival of other neutrophils, particularly in sites where the cells are in close proximity as in the marrow. Self destruction of post-mitotic neutrophils in marrow may thus represent an-other level at which regulation of cell production展开更多
文摘To clarify the hematopoietic potential of various sub-classes of human hematopoietic progenitor cells, we used a multicolor staining protocol in conjunction with anti-CD34 and -CD38 McAb. We characterized two cell fractions in CD34+cells with or without CD38 expression. A clonogenic assay showed that most CFC were present in CD34+CD38+ population. Morphologic analysis showed that blast-like cells were more enriched in the CD34+CD38 fraction. To clarify the biologic differences between both fractions, we examined the more primitive progenitor cell function by assessing long-term culture-initiating cells (LTC-IC) on the stromal cells. At the first two weeks, more CF.C harvested from the culture in the fractions initiated with both populations. However, more LTC-IC were present during weeks 4 to 12 in the CD34+CD38- population. These results indicate the primitive progenitors are more enriched in CD34+CD38 population than in CD34+CD38+ cells.
文摘Antitumor chemotherapy is often limited by hematopoietic toxicity.TO attenuate themyelosuppressive effects of chemotherapy, successfulengraftment of unmodified fetal liver cells (FLCs) hasbeen achieved across allogeneic barriers in a number ofanimal models and patients, GM-CSF is a
文摘Previous reports indicated that the difference ofMHC-Ⅰ and Ⅱ between donor and recipient was criticalfor the occurrence of allogenic rejection or graft versushost reaction (GVHR). This study aimed at looking forthe possibility of reducing the risk of immunologicalrejection through transplantation tolerance induced bytransduction of MHC gene. In the present study, wetransferred donor mice MHC class I gene (H-2k^b gene)
文摘Stem cell factor (SCF) is a novel growth factor thatinfluences the growth and development of hematopoieticcells, germ cells and melanocytes. To explore
文摘Liposomes have several advantages over viral vectorsfor gene delivery both in vitro and in vivo. However,few data are available concerning gene transfer intohematopoietic stem cells. In order to explore theefficiency and the stability of expression of gene transferinto hematopoietic stem cells, we have transduced twomarker genes (Neo^R and Lac Z) co-transfer into humanbone marrow CD34^+ hematopoietic stem cells
文摘The effect of recombinant human interleukin-6 ( rhIL-6) on the early phase of post-irradiation hematopoietic recovery ( mainly CFU-S, CFU-GM and CFU-E) has been studies in 6.5 Gy gamma-irradiated C57BL / 6J mice, which had been given subcutaneous injection of rhIL-6 b. i. d. at 10, 100, 200, 500 and 1 000μg/kg/d for 4 or 6 consecutive days started from 30 min after irradiation. It was shown that the effect of IL-6 on the early phase of post-irradiation hematopoietic recovery is dose-dependent and diphasic. At the doses of 200-1000μ/kg/d, IL-6 had a favorable effect on post-irradiation hematopoietic recovery expressed as increse of spleen weight and CFU-S, and elevation of CFU-GM yield and total CFU-GM content in femur. Administration of relatively low doses of IL-6(10-100 μg/kg/d) suppressed postirradiation hematopoietic regeneration. High a dose (1 000μg/kg/d) did not show any stronger stimulative effect on post-irradiation hematopoietic reconstitution. There is an optimal range of IL-6 dose for
文摘Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hemo-lytic disease in which there is a stem cell disorder of clonal nature. Previous studies have demonstrated that the numbers of burst-forming units-erythroid (BFU-E) and colony-forming units-granulocyte / macrophage (CFU-GM) from bone marrow of PNH patients growing in the medium containing PHA-LCM from the normai donors were more reduced than those of normai bone marrow. The purpose of present study was to investigate if PNH lymphocytes are defective in supporting hematopoiesis in vitro. PHA-LCM from PNH blood was added to the culture medium for the growth of PNH and normai BFU-E and CFU-GM. The numbers of PNH bone marrow BFU-E and CFU-GM in the medium containing PHA-LCM from PNH blood were less than those from normai blood; the numbers of normai bone marrow BFU-E and CFU-GM grown in the medium containing PHA-LCM from PNH blood were more decreased than those from normai blood. The results suggest that diminished numbers of PNH bone marrovv BFU-
基金Key Program of Capital Development Foundation(编号2007-2040)
文摘In order to study the epidemiological characteristics of cytomegalovirus(CMV)infection in allogeneichematopoietic stem cell transplantation(allo-HSCT)recipients by means of plasma real time quantitative polymerasechain reaction(RQ-PCR),141 adult patients undergoing allo-HSCT between January 2008 and June 2010 were seriallymonitored by RQ-PCR for detecting CMV and guiding the preemptive therapy followed up to 180 days post-HSCT.Theresults showed that the incidence of CMV infection and CMV pneumonia was 81.5%and 2.9%respectively,whichmainly occurred within 2 months post-HSCT.Single-therapy with ganciclovir(GCV)for 63 patients or foscarnet 6patients was performed for preemptive therapy.The total efficacy was 87.8%,and the response patterns were different.CMV infection was more frequent in female patients(P=0.044),and those with aGVHD(P=0.043),using ATG orbasiliximab in conditioning regimens(P=0.049),as well as earlier in patients using ATG or basiliximab or those withaGVHD(P=0.007;P=0.000).The aGVHD,maximum load,positive times of CMV-DNA detection and therapyduration all correlated with the efficacy(P<0.05).It is concluded that the incidence of CMV infection is still high afterHSCT.Plasma RQ-PCR assay for CMV-DNA shows a strong correlation with the clinical outcome of CMV infection,which is useful and suitable for management of CMV infection in HSCT.
文摘To elucidate the feasibility of retroviral mediatedtransfer of multidrug resistance gene (mdrl) intohematopoietic stem/progenitor cells, the CD34^+ cellswere isolated from bone marrow by using a high-gradient.magnetic cell sorting system, and the efficiency ofretroviral mediated gene transfer was studied.
文摘It has been demonstrated that tbe critical role ofbone marrow stromal cells (HMSCs ) is to sustain theselfrenewal of pluripotent hematopoietic stem cells andmaintain the homeostasis of bone marrow hematopoiesismicroenvironment. BMSC progenitor can differentiateinto several clements including macrophages, endothelialcells, fibroblasts and some other cells. Almost
文摘Cancer gene therapy over the past several yearshas involved the introduction of genes intohematopoietic cells for: (1) protecting the normalcells from the side effects of chemotherapy; (2)introduction of genes into the neoplastic cells for
文摘Retroviral mediated gene transfer of humanglobin gene into hematopoietic stem cells is apromising approach for thalassemia gene therapy.Major problem of the transferred globin gene was lowlevel expression of the gene with its proximal cis-acting sequence. The locus control region (LCR) ofthe human β-globin gene cluster consists of four majorDNase I hypersensitive sites (HS). When linked
文摘Chemotherapy is one of the curative treatmentmodality for several types of tumors. However, thedosage of antitumor drugs was limited by cytotoxicityof drugs to normal tissues, especially for bone marrow(myelosuppression). Drug resistantce gene (hMDRI,hDHFR etc.) is expressed in many normal tissues. Itsover-expression in tumor cells give rise to the tumorresistant to many drugs. But, if we transfer the drugresistance gene into the normal hematopoietic stemcell (HSC), the expression of the foreign gene
文摘In order to increase the expression of mdrl genetransfectcd using retroviral vcctors, we constructedtwo retroviral vectors (LmdrlSN and MSCV-mdrl)containing mdrl cDNA and compared the expressionof mdrl gene with another vector pHamdrl/A(Pastan et al. Proc Natl Acad Sci USA 85: 4486,1986) in NIH.3T3 cells. The LmdrlSN was made
基金Rothrock Research Fund in Hematology, The authors appreclate the assistance of supervising technicina, Linda Russ.Ulrica Stenheimer-Caudle, Sandra Peffly, and Marie Hyde.
文摘An extract (G-INH) made from mature human granulocytes freshly isolated from normai blood causes human neutrophils to undergo apoptosis in vitro as shown by morphologic changes and by the typical ladder pattern of small DNA fragments noted on agarose gel electrophoresis of isolated DNA. Apoptosis occurs in from 20% to 30% of neutrophils over 24 hours of culture in vitro and the addition of G-INH to the medium causes a dose-related increase in the incidence of apoptosis. Heating G-INH at 60t for 30 minutes does not destroy its capacity to induce apoptosis but GM-CSF, G-CSF, and to a lesser extent IL-1β, antagonize this action. IL-3 does not diminish G-INH induced apoptosis of neutrophils. Substances, released from, mature neutrophils may participate in regulating the survival of other neutrophils, particularly in sites where the cells are in close proximity as in the marrow. Self destruction of post-mitotic neutrophils in marrow may thus represent an-other level at which regulation of cell production
