Objective:To investigate the effects of soluble factors secreted by acute myeloid leukemia(AML) cells on the phenotypical and functional properties of DCs derived from normal mononuclear cells. Methods:Mononuclear cel...Objective:To investigate the effects of soluble factors secreted by acute myeloid leukemia(AML) cells on the phenotypical and functional properties of DCs derived from normal mononuclear cells. Methods:Mononuclear cells were cul-tured with interleukin-4(IL-4) and granulocyte-macrophage colony-stimulating factor(GM-CSF),in the presence or absence of 24 h culture supernatants from fresh primary AML cells,to generate immature DCs. The maturation of DCs was induced by cytokines IL-1beta,IL-6,tumor necrosis factor-alpha(TNF-alpha),and prostaglandin-2(PGE-2). The phenotypic alterations of DCs and DCs-primed CD4+ T cells were evaluated using flow cytometry. Precursor frequency(PF) was calculated to monitor the allostimulatory effects of DCs on CD4+ and CD8+ T cells. Results:AML cell supernatant-treated DCs showed significantly lower expression of co-stimulatory molecules CD80 and CD86,and reduced response to cytokines IL-1beta,IL-6,TNF-alpha,and PGE-2. The allostimulatory effects of AML cell supernatant-treated DCs on CD4+ and CD8+ T cells were significantly lower than those of normal mature DCs [PF:(1.8 ± 0.5)% vs.(5.2 ± 1.6)% for CD4+ T cells,(2.1 ± 0.6)% vs.(6.5 ± 2.0)% for CD8+ T cells,P < 0.01]. These AML supernatant-induced DCs could also induce allogeneic CD4+ T cells to differentiate into CD4+CD25high T cells,which had immunophenotyping characteristics of regulatory T cells,i.e. they expressed Foxp3 but not active maker CD69. Conclusion:This study demonstrates that soluble factors secreted by AML cells can inhibit development and functions of DCs. In addition,AML supernatant-induced DCs can induce the generation of CD4+CD25high T cells from CD4+ T cells,which may be a mechanism of increased prevalence of CD4+CD25high regulatory T cells and immune dysfunction in AML patients.展开更多
Dendritic cells (DCs), as the most potent antigen presenting cells in vivo, are central to generating specific immune responses and can be used as important vectors in antitumour immunity.~1 DCs loaded with tumour ant...Dendritic cells (DCs), as the most potent antigen presenting cells in vivo, are central to generating specific immune responses and can be used as important vectors in antitumour immunity.~1 DCs loaded with tumour antigens can induce specific antitumour response in cytotoxic T lymphocytes (CTL). P-glycoprotein (P-gp), a product of mdr1 gene, widely overexpressed in many multidrug resistant (MDR) tumour cells, is a main mechanism that makes tumour cells escape from death induced by chemotherapeutic agents. However, this drug efflux protein may also be regarded as the antigen to active cellular immunity. It has been demonstrated that展开更多
Objectives To assess the feasibility and efficacy of eliciting leukemia-specific T-cell responses in syngeneic mice in vitro and in vivo using dendritic cells (DCs) pulsed with total RNA from leukemia cells.Methods DC...Objectives To assess the feasibility and efficacy of eliciting leukemia-specific T-cell responses in syngeneic mice in vitro and in vivo using dendritic cells (DCs) pulsed with total RNA from leukemia cells.Methods DCs generated from bone marrow culture in vitro in the presence of combined cytokines were pulsed with cellular total RNA isolated from cultured L615 cells by cationic lipid 1,2-dioleoyloxy-3-(trimethylammonium) propane (DOTAP). T-cell responses were evaluated by in vitro proliferation, and cytotoxicity assay. And in vivo immune protection and proghosis of mice with leukemia were studied.Results DCs pulsed with total RNA isolated from cultured L615 cells (DCs/RNA) were remarkably effective in stimulating L615-specific T-cell response in vitro, but did not cross-react with other leukemia cells from syngeneic mice. Vaccination of naive mice with viable DCs/RNA vaccine was able to partly protect from challenge with a lethal dose of live L615 cells, leading to low leukemia incidence and overall survival prolongation. Statistically significant survival was also observed in a low lethal dose of L615-bearing mice that received treatment using viable DCs / RNA vaccine alone, suggesting that systemic administration of IL-2 could enhance the anti-tumor efficacy of leukemia RNA/DCs vaccine.Conclusions These data support the use of DCs/RNA vaccine as a feasible and effective route to elicit leukemia immunity against unidentified leukemia-associated antigens for treatment of leukemia-bearing animals.展开更多
Introduction:Mature plasmacytoid dendritic cells(pDCs)proliferation associated with myeloid neoplasms(MPDMN)are recognized as a neoplasm related to fully differentiated pDCs.Although it has been reported for many year...Introduction:Mature plasmacytoid dendritic cells(pDCs)proliferation associated with myeloid neoplasms(MPDMN)are recognized as a neoplasm related to fully differentiated pDCs.Although it has been reported for many years,the genomic landscape of MPDMN is poorly understood.Methods:We reported two patients who developed acute myeloid leukemia(French-American-British M5 subtype)coexisted with immunophenotypically mature pDCs proliferation,which fit the diagnosis of MPDMN.We sorted pDCs from myeloid blasts by flow cytometry and performed whole-exome sequencing and RNA sequencing of the two cell populations,respectively.Results:The immunophenotypes of pDCs in both patients were positive for CD123bri,HLA-DR,CD4,CD303,CD304,and negative for CD56,CD34,CD117,and TdT.The variant allele frequency of gene mutations in myeloid blasts and pDCs were similar.The expression data showed myeloid blasts clustered tightly with hematopoietic stem cells,and pDCs from patients clustered tightly with granulocyte-monocyte progenitors/common myeloid progenitor,rather than with pDCs from the GEO platform.Conclusion:Our study suggested that pDCs derived from the leukemic clone,evidenced by a shared mutation profile and similar transcriptional signatures between pDCs and concurrent myeloid blasts.展开更多
In general,acute myeloid leukemia(AML)is an aggressive and heterogeneous disease that is characterized by rapid cellular proliferation and high mortality.One of the mutations related to AML is the Flt3-ITD mutation,wh...In general,acute myeloid leukemia(AML)is an aggressive and heterogeneous disease that is characterized by rapid cellular proliferation and high mortality.One of the mutations related to AML is the Flt3-ITD mutation,which is found in approximately 25%of patients.In this mini-review,we investigate the function of dendritic cells and T cells based on Flt3-ITD mutation and immune evasion as a result of this abnormality.Finally,we discuss some AML therapeutic strategies,including targeting Flt3 on DCs and TIM-3 on T cells as immune receptors to treat this hematopoietic malignancy.展开更多
To investigate the antitumor activity of IL 12, the induction of differentiation of IL 12 was observed using erythroleukemia cells (FBL 3) as model. After incubation with 200 U/mL IL 12 for 48 h, DNA synthesis of FBL ...To investigate the antitumor activity of IL 12, the induction of differentiation of IL 12 was observed using erythroleukemia cells (FBL 3) as model. After incubation with 200 U/mL IL 12 for 48 h, DNA synthesis of FBL 3 cells in S phase decreased significantly; the expression of CD14 which is the specific marker of monocyte increased, the rate of NBT + cells was apparently higher than that of the untreated FBL 3 cells. After treating FBL 3 cells with IL 12 for 72 h, the expression of 33D1 and NLDC145 which are the specific markers of dendritic cells increased markedly, the surface molecules such as MHC II,B7 1, B7 2, and VCAM 1 were up regulated; morphological observation showed two kinds of cells: some cells had a ruffled surface and plentiful lysosome; the others had many dendritic projections on the surface, and contained numerous mitochondria. Functionally, the IL 12 treated FBL 3 cells could apparently stimulate the proliferation of allogeneic and autologous T lymphocytes, and improve the specific cytotoxic activity of CTL on FBL 3 cells. These results indicated that erythroleukemia cells were induced by IL 12 to differentiate into the monocytes and dendritic cells, then exhibited the antigen presenting function. The data outline a new mechanism for IL 12 to treat leukemia.展开更多
基金grants from the National Outstanding Young Investigator Program (30225038)Anhui Provincial Natural Science Foundation (070413094)+1 种基金Scientific Research Fund of Anhui Provincial Education Department (2006KJ072C)Science and Technological Fund of Anhui Province for Outstanding Youth.
文摘Objective:To investigate the effects of soluble factors secreted by acute myeloid leukemia(AML) cells on the phenotypical and functional properties of DCs derived from normal mononuclear cells. Methods:Mononuclear cells were cul-tured with interleukin-4(IL-4) and granulocyte-macrophage colony-stimulating factor(GM-CSF),in the presence or absence of 24 h culture supernatants from fresh primary AML cells,to generate immature DCs. The maturation of DCs was induced by cytokines IL-1beta,IL-6,tumor necrosis factor-alpha(TNF-alpha),and prostaglandin-2(PGE-2). The phenotypic alterations of DCs and DCs-primed CD4+ T cells were evaluated using flow cytometry. Precursor frequency(PF) was calculated to monitor the allostimulatory effects of DCs on CD4+ and CD8+ T cells. Results:AML cell supernatant-treated DCs showed significantly lower expression of co-stimulatory molecules CD80 and CD86,and reduced response to cytokines IL-1beta,IL-6,TNF-alpha,and PGE-2. The allostimulatory effects of AML cell supernatant-treated DCs on CD4+ and CD8+ T cells were significantly lower than those of normal mature DCs [PF:(1.8 ± 0.5)% vs.(5.2 ± 1.6)% for CD4+ T cells,(2.1 ± 0.6)% vs.(6.5 ± 2.0)% for CD8+ T cells,P < 0.01]. These AML supernatant-induced DCs could also induce allogeneic CD4+ T cells to differentiate into CD4+CD25high T cells,which had immunophenotyping characteristics of regulatory T cells,i.e. they expressed Foxp3 but not active maker CD69. Conclusion:This study demonstrates that soluble factors secreted by AML cells can inhibit development and functions of DCs. In addition,AML supernatant-induced DCs can induce the generation of CD4+CD25high T cells from CD4+ T cells,which may be a mechanism of increased prevalence of CD4+CD25high regulatory T cells and immune dysfunction in AML patients.
文摘Dendritic cells (DCs), as the most potent antigen presenting cells in vivo, are central to generating specific immune responses and can be used as important vectors in antitumour immunity.~1 DCs loaded with tumour antigens can induce specific antitumour response in cytotoxic T lymphocytes (CTL). P-glycoprotein (P-gp), a product of mdr1 gene, widely overexpressed in many multidrug resistant (MDR) tumour cells, is a main mechanism that makes tumour cells escape from death induced by chemotherapeutic agents. However, this drug efflux protein may also be regarded as the antigen to active cellular immunity. It has been demonstrated that
基金National Natural Science Research Foundation (No. 9770825).
文摘Objectives To assess the feasibility and efficacy of eliciting leukemia-specific T-cell responses in syngeneic mice in vitro and in vivo using dendritic cells (DCs) pulsed with total RNA from leukemia cells.Methods DCs generated from bone marrow culture in vitro in the presence of combined cytokines were pulsed with cellular total RNA isolated from cultured L615 cells by cationic lipid 1,2-dioleoyloxy-3-(trimethylammonium) propane (DOTAP). T-cell responses were evaluated by in vitro proliferation, and cytotoxicity assay. And in vivo immune protection and proghosis of mice with leukemia were studied.Results DCs pulsed with total RNA isolated from cultured L615 cells (DCs/RNA) were remarkably effective in stimulating L615-specific T-cell response in vitro, but did not cross-react with other leukemia cells from syngeneic mice. Vaccination of naive mice with viable DCs/RNA vaccine was able to partly protect from challenge with a lethal dose of live L615 cells, leading to low leukemia incidence and overall survival prolongation. Statistically significant survival was also observed in a low lethal dose of L615-bearing mice that received treatment using viable DCs / RNA vaccine alone, suggesting that systemic administration of IL-2 could enhance the anti-tumor efficacy of leukemia RNA/DCs vaccine.Conclusions These data support the use of DCs/RNA vaccine as a feasible and effective route to elicit leukemia immunity against unidentified leukemia-associated antigens for treatment of leukemia-bearing animals.
基金was supported in part by State Key Program of National Natural Science of China(81830005)J.W.,National Natural Science Foundation of China(81770181)+3 种基金J.W.,National Key Research and Development Program of China(2019YFC0840605)Y.M.,Tianjin Natural Science Foundation(18JCZDJC45000)H.W.,CAMS Innovation Fund for Medical Sciences(2020-I2M-C&T-B-084)H.W.Funders had no role in the study design,analyses,or decision to publish.
文摘Introduction:Mature plasmacytoid dendritic cells(pDCs)proliferation associated with myeloid neoplasms(MPDMN)are recognized as a neoplasm related to fully differentiated pDCs.Although it has been reported for many years,the genomic landscape of MPDMN is poorly understood.Methods:We reported two patients who developed acute myeloid leukemia(French-American-British M5 subtype)coexisted with immunophenotypically mature pDCs proliferation,which fit the diagnosis of MPDMN.We sorted pDCs from myeloid blasts by flow cytometry and performed whole-exome sequencing and RNA sequencing of the two cell populations,respectively.Results:The immunophenotypes of pDCs in both patients were positive for CD123bri,HLA-DR,CD4,CD303,CD304,and negative for CD56,CD34,CD117,and TdT.The variant allele frequency of gene mutations in myeloid blasts and pDCs were similar.The expression data showed myeloid blasts clustered tightly with hematopoietic stem cells,and pDCs from patients clustered tightly with granulocyte-monocyte progenitors/common myeloid progenitor,rather than with pDCs from the GEO platform.Conclusion:Our study suggested that pDCs derived from the leukemic clone,evidenced by a shared mutation profile and similar transcriptional signatures between pDCs and concurrent myeloid blasts.
文摘In general,acute myeloid leukemia(AML)is an aggressive and heterogeneous disease that is characterized by rapid cellular proliferation and high mortality.One of the mutations related to AML is the Flt3-ITD mutation,which is found in approximately 25%of patients.In this mini-review,we investigate the function of dendritic cells and T cells based on Flt3-ITD mutation and immune evasion as a result of this abnormality.Finally,we discuss some AML therapeutic strategies,including targeting Flt3 on DCs and TIM-3 on T cells as immune receptors to treat this hematopoietic malignancy.
文摘To investigate the antitumor activity of IL 12, the induction of differentiation of IL 12 was observed using erythroleukemia cells (FBL 3) as model. After incubation with 200 U/mL IL 12 for 48 h, DNA synthesis of FBL 3 cells in S phase decreased significantly; the expression of CD14 which is the specific marker of monocyte increased, the rate of NBT + cells was apparently higher than that of the untreated FBL 3 cells. After treating FBL 3 cells with IL 12 for 72 h, the expression of 33D1 and NLDC145 which are the specific markers of dendritic cells increased markedly, the surface molecules such as MHC II,B7 1, B7 2, and VCAM 1 were up regulated; morphological observation showed two kinds of cells: some cells had a ruffled surface and plentiful lysosome; the others had many dendritic projections on the surface, and contained numerous mitochondria. Functionally, the IL 12 treated FBL 3 cells could apparently stimulate the proliferation of allogeneic and autologous T lymphocytes, and improve the specific cytotoxic activity of CTL on FBL 3 cells. These results indicated that erythroleukemia cells were induced by IL 12 to differentiate into the monocytes and dendritic cells, then exhibited the antigen presenting function. The data outline a new mechanism for IL 12 to treat leukemia.
基金this study was supported by a grant from the Shaanxi social development key fund (No.2007k0902)supported byShaanxi Three-Five talent engineering fund(2009)
