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OBJECTIVE@#To investigate the effects of dasatinib on the maturation of monocyte-derived dendritic cells (moDCs) derived from healthy donors (HDs) and chronic myelogenous leukemia (CML) patients.@*METHODS@#Peripheral blood mononuclear cells (PBMCs) were isolated from HDs (n=10) and CML patients (n=10) who had got the remission of MR4.5 with imatinib treatment. The generation of moDCs from PBMCs was completed after 7 days of incubation in DC I culture medium, and another 3 days of incubation in DC II culture medium with or without 25 nmol/L dasatinib. On the 10th day, cells were harvested and expression of molecules of maturation related marker were assessed by flow cytometry. The CD80+CD86+ cell population in total cells was gated as DCs in the fluorescence-activated cell storting (FACS) analyzing system, then the expression of CD83, CD40 or HLA-DR in this population was analyzed respectively.@*RESULTS@#The proportion of CD80+CD86+ cells in total cells didn't show a statistical difference between HD group and patient group (89.46%±9.70% vs 87.39%±9.34%, P=0.690). Dasatinib significantly enhanced the expression of the surface marker CD40 (P=0.008) and HLA-DR (P=0.028) on moDCs derived from HDs compared with the control group, while the expression of CD83 on moDCs didn't show a significant difference between dasatinib group and the control group (P=0.428). Meanwhile, dasatinib significantly enhanced the expression of the surface marker CD40 (P=0.023), CD83 (P=0.038) and HLA-DR (P=0.001) on moDCs derived from patients compared with the control group.@*CONCLUSION@#For CML patients, the same high proportion of moDCs as HDs can be induced in vitro, which provides a basis for the application of DC-based immunotherapy strategy. Dasatinib at the concentration of 25 nmol/L can efficiently promote the maturation of moDCs derived from HDs and CML patients in vitro. Dasatinib shows potential as a DC adjuvant to be applied in DC-based immunotherapy strategies, such as DC vaccine and DC cell-therapy.
Subject(s)
Humans , Cell Differentiation , Cells, Cultured , Dasatinib/pharmacology , Dendritic Cells , HLA-DR Antigens/pharmacology , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/metabolism , Leukocytes, Mononuclear , MonocytesABSTRACT
Objective: To explore the expressions of Foxp3 ' Tregs and CDllc mDCs in the colorectal cancer (CRC) tissue and their infiltrations in the tumor draining lymph node (TDLN) tissue, and to explore the clinical significances. Methods: Fifty-two samples of surgical resection of the CRC patients were collected The expressions of Foxp3 + Tregs and CDllc mDCs in the specimens of the CRC patients and TDLN tissue were detected by immunohistochemistry, and the relationship between the expression frequencies of Foxp3 + Tregs and CD11c+ mDCs and the clinicopathological characteristics of the patients were discussed Results: The positive expression frequency of Foxp3 + Tregs in cancer tissue of the CRC patients was higher than that in adjacent normal mucosa tissue (P0. 05). Conclusion: The occurrence and development of CRC are related to the changes of Foxp3 Tregs and CDllc mDCs expression frequencies in the local microenvironment of cancer tissue. Foxp3+ Tregs and CD11c+ mDCs play a role in inhibiting the cellular immunity in tumor microenvironment.
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Blastic plasmacytoid dendritic cell neoplasm is a rare hematological malignancy derived from immature plasmacytoid dendritic cells. The tumor cells have an immature blastic appearance, and diagnosis is based on the expression of CD4, CD56 y CD123 in the absence of other lymphoid, natural killer, or myeloid antigens. The majority of affected individuals are older people with a mean age of 66 years. Male to female ratio is approximately 3:1. Common presentation includes cutaneous lesions followed by tumor dissemination. Treatment with conventional chemotherapy is ineffective and allogeneic hematopoietic stem cell transplantation is required to achieve remission. We report three male patients, aged 23, 27 and 51 years with the disease. All had multiple, infiltrated pink plaques and nodules on the skin of their face, neck and thorax, measuring 1 to 12 cm in diameter. All tumors were histologically characterized by a monotonous proliferation of medium size cells with blastic features. Tumor cells were positive for CD123, CD56, CD4 and CD7 in all cases. After a mean of follow-up of 14.6 months, one patient died of the disease, one patient is alive and the disease relapsed after 17 months of remission and one patient is alive with no evidence of the disease.
Subject(s)
Humans , Male , Adult , Middle Aged , Young Adult , Dendritic Cells/pathology , Hematologic Neoplasms/pathology , Skin/pathology , Biopsy , Bone Marrow/pathology , Immunohistochemistry , Fatal OutcomeABSTRACT
Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare, clinically aggressive hematologic malignancy that most commonly manifests as cutaneous lesions with or without bone marrow involvement and leukemic dissemination. The demonstration of tumor cells with the characteristic immunophenotype with expression of CD56, generally CD4 and dendritic cell antigens (CD123, cyTCL-1, HLA-DR), in the absence of myeloid or lymphoid lineage markers is required for the diagnosis. Responses to chemotherapy are initially satisfactory, with frequent systemic and central nervous system relapses. We report a 24 year-old male with BPDCN, initially diagnosed and treated as non-Hodgkin CD4+ T-cell lymphoma, with initial complete remission who evolved with early central nervous system relapse. A second attempt of chemotherapy failed and the patient died two months later.
Subject(s)
Humans , Male , Young Adult , Dendritic Cells/pathology , Central Nervous System Neoplasms/secondary , Hematologic Neoplasms/pathology , Remission Induction , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Immunophenotyping , Fatal Outcome , Disease Progression , Hematologic Neoplasms/drug therapyABSTRACT
AIM:To investigate the role of mesenchymal stem cell-induced regulatory dendritic cells ( MSC-DCregs) in mouse acute graft-versus-host disease( aGVHD) model.METHODS: Bone marrow cells from BALB/c ( H-2 d ) mice were isolated and were induced to differentiate into DCs.The DCs were selected by flow cytometry, and after 10 d co-culture with MSCs, they were induced to be MSC-DCregs.Male 8-week-old C57BL/6 (H-2b) mice were used as do-nor mice.The female 8-week-old BALB/c (H-2d) mice, who had received 100 cm source-skin distance, 30 cm ×30 cm radiation field, 700 cGy total body irradiation (TBI) pretreatment were used as recipient mice.The recipients were divided into 5 groups:control group, TBI group ( injected with medium only) , bone marrow transplantation group ( injected with 1 ×107 bone marrow cells), aGVHD group (injected with 1 ×107 bone marrow cells and 1 ×107 spleen cells), and MSC-DCregs group (injected with 1 ×107 bone marrow cells, 1 ×107 spleen cells and 1 ×106 MSC-DCregs).The white blood cell count, recipients’ chimerism, clinical evaluation of aGVHD, survival analysis and pathological changes were deter-mined.RESULTS:Hematopoieic recovery was seen at 10 d after transplantation.The recipients’ chimerism was parallel to the donors’ at 30 d.The median survival time of the mice in aGVHD group and MSC-DCregs group was 27 d and 33 d, and the survival rates at 30 d were 20% and 100% (P<0.01), respectively.The clinical scores of the mice in MSC-DCregs group were lower than those in aGVHD group ( P<0.01) .Moreover, the pathological changes in the skin and liver of the mice in MSC-DCregs group were less serious than those in aGVHD group.CONCLUSION: The MSC-DCregs in-duce an aGVHD tolerance in vivo, and further research of its mechanism is still in great necessary.
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Objective To investigate the biological effect of GM-CSF on splenic dentritic cells (DCs)when used to treat severe sepsis and the influence on the prognosis.Methods All 160 male Kunming mice were randomly(random number)divided into four groups:control group(n =50),the mice didnt receive special treatment; CLP group(n =42),the mice underwent cecal ligation and puncture;conventional treatment group(n =34),the mice received intraperitoneal injection of 5 mg/kg ceftriaxone at 12,24,36,48,60,72 and 84 h after surgery; GM-CSF treatment group(n =34),the mice received hypodermic injection of 20 μg/kg GM-CSF besides ceftriaxone at 12,36,60,and 84 h after surgery.The mice were sacrificed at 0,12,24,48 and 72 h after CLP by cervical dislocation.The amount of splenic DCs and apoptosis rate were measured by flow cytometry,and the serum concentrations of IL-12 were detected by ELISA in each group.Meanwhile the survival prognosis was observed at 96 h.Results At every time point,the apoptosis of splenic DCs increased and the amount markedly reduced in severe sepsis(P ＜0.05),the serum concentration of IL-12 increased on an one-way curve type(P ＜0.05).The treatment of cephalosporin exacerbated the loss of DCs(P ＜ 0.05),while hadnt any effect on IL-12(P ＞ 0.05).GMCSF treatment increased the amount of DCs(P ＜ 0.05),and decreased IL-12 concentrations(P ＜ 0.05).The OR of GM-CSF was 0.079 computed by Logist regression analysis.Conclusions GM-CSF treatment severe sepsis can increase the amount of splenic DCs,decrease serum levels of IL-12,and improve prognosis.
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Objective To investigate the killing effect of donor NK cells on recipient dentritic cells (DCs) by blocking KIR2DL1 and KIR2DL3 with monoclonal antibodies CD158a and CD158b,and the possible mechanism about donor NK cells reducing the incidence of graft versus host disease (GVHD).Methods SSP-PCR was used to examine the genotypic makeup of KIR in 15 pairs of patients and their HLA-identical sibling hematopoietic stem cell transplantation (HSCT) donors in our department.Peripheral blood mononuclear cells (PBMC) from 15 pairs of patients and their donors were extracted and preserved,and donor NK cells were isolated by DYNABEADS UNTOUCHED HUMAN NK sorting kit.Methyl thiazolyl tetrazolium (MTT) reduction assay was used to detect the killing activity of donor NK cells against recipients DCs.To observe the different NK killing activity before and after KIR receptors blockade,the anti-CD158a and CD158b monoclonal antibodies were used to block KIR inhibitory receptors.Results The killing effect of donor NK cells was significantly enhanced after inhibitory receptor KIRs blockades,the killing effect of NK cells in HVG KIR ligandmismatching pattern group was significantly higher than GVH or KIR receptor-ligand matched pattern group.The killing effect became stronger when the dose of antibody was increased.ConclusionIt is demonstrated in vitro that the killing effect of alloreactive NK cells could be enhanced by KIR functional modification,and the infusion of alloreactive NK cells could be used as a new strategy of immunotherapy for GVHD.
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To investigate effects of rhIL-17 on growth and development of mouse bone marrow progenitors andhuman cord blood LD34~+ stem cells. Methods: Mouse bone marrow progenitors were isolated by routine protocol, and CD34~+ stem cells were isolated from normal human cord blood by Mini-MACS, then cultured with rhIL-17 and/or GM-CSF/IL-4. The phenotypes of the cells were analyzed by FACS, IL-12 level was analyzed by ELISA and T cell stimulating activity in allo-MLR was determined by [~3H]-TdR incorporation. Results: Expression of MHC class II molecules and B7-2 on the surface of immature DC derived from mouse bone marrow progenitors was up-regulated by IL-17. The capacity of the cells to secrete IL-12 and their T cell stimulating activity were also enhanced. The cells showed the characteristics of mature DC. After cultured with IL-17 for 9 days, the number of CD34~+ stem cells increased by 2 times. The phenotypes of some cells were CDla~(high), B7-2~(high), and HLA-DR~(lwo). The cells could stimulate allo geneic T cells to proliferate but their capacity was lower than that of the cells cultured with IL-17 combined with GM-CSF. The cells cultured with IL-17 and GM-CSF proliferated markedly and the rate of CDla~+ and B7-2~+ cells increased significantly. The T cell stimulating activity of cells was also augmented. Conclusion: IL-17 could promote DC derived from mouse bone marrow progenitors to mature. When combined with GM-CSF, IL-17 could induce human CD34~+ stem cells not only to proliferate markedly but also to show characteristics of DC, indicating that CD34~+ stem cells might differentiate to DC by IL-17.