Irradiation of peripheral blood mononuclear cells with 7.5 Gy X-rays prior to donor lymphocyte infusion inhibits proliferation while preserving cytotoxicity, and improves the effectiveness of HSCT in patients with hematological malignancies.

The aim of the present study was to explore the effect of different X-ray doses on the proliferation and cytotoxic activity of peripheral blood mononuclear cells (PBMCs), particularly lymphocytes, in order to assess whether this reduces the incidence of graft vs. host disease (GVHD) while preserving the graft vs. tumor (GVT) effect in patients with hematological malignancies following hematopoietic stem cell transplantation (HSCT).

Allogeneic Compact Bone-Derived Mesenchymal Stem Cell Transplantation Attenuates the Severity of Idiopathic Pneumonia Syndrome in a Murine Bone Marrow Transplantation Model.

Background/aims: Idiopathic pneumonia syndrome (IPS) is a serious and life-threatening lung complication following allogeneic hematopoietic stem cell transplantation (allo-HSCT) and currently no effective therapies exist. This study was designed to determine whether transplantation of allogeneic murine compact bone derived- mesenchymal stem cells (CB-MSCs) could prevent the development of IPS.

Methods: We tested the effects of CB-MSCs transplantation on IPS using an established murine model of C57BL/6 (H-2b)→BALB/c (H-2d).

Prophylaxis of acute graft-versus-host disease by CCR5 blockade combined with cyclosporine A in a murine model.

Objective: One characteristic feature of graft-versus-host disease (GVHD) is lymphocytes' trafficking and recruitment to target tissues, and CCR5 plays a key role in the process. Thus, blockade of lymphocytes' chemotaxis may attenuate GVHD.

Methods: We tested the effects of CCR5 blockade using an established murine model.

In vitro immunological effects of blocking CCR5 on T cells.

Blockade of CC chemokine receptor 5 (CCR5) by maraviroc may induce immunological changes independent of its antiviral effects and may have immunoregulation properties. This study was designed to determine the effects of blocking CCR5 on human activated T cells in vitro and investigate the potential immunological mechanisms. Human CD3+ T cells were purified from peripheral blood mononuclear cells and then activated by cytokines.

Silencing HCCR2 expression inhibits the proliferation of leukemia cells by inducing apoptosis and promoting cell cycle arrest.

The human cervical cancer oncogene (HCCR2) has been found to be overexpressed in a variety of human malignant tumors cells, and its function is related to cell cycle progression and survival. However, the molecular mechanisms of action of HCCR2 in leukemia remain unclear. In this study, we used the RNA interference strategy to investigate the effects of HCCR2 knockdown in the K562 leukemia cell line, and to explore the potential mechanisms involved.

Bortezomib regulates the chemotactic characteristics of T cells through downregulation of CXCR3/CXCL9 expression and induction of apoptosis.

The chemotactic movement of T lymphocytes mediated by chemokines and their receptors plays an important role in the pathogenesis of graft-versus-host disease (GVHD) post-allogeneic hematopoietic stem cell transplantation (allo-HSCT). CCR7 and CXCR3 are two receptors associated with the development of GVHD. Bortezomib, a proteasome inhibitor, was recently found to prevent GVHD in a mouse model and to decrease the production of Th1 cytokines.

[Bone marrow-derived mesenchymal stem cells regulate the proliferation and activity of natural killer cells].

This study was aimed to explore the effect of bone marrow-derived mesenchymal stem cells (MSC) on proliferation and activity of natural killer (NK) and NK-T cells. MSC was co-cultured with peripheral mononuclear cells from healthy donors in presence of IL-2, phytohemagglutinin (PHA) and mouse anti-human CD3 McAb (culture condition known to expand NK cells). The ratio of NK cells and NK-T cells was measured by flow cytometry and the effect of MSC on killing activity of NK cells against K562 cells was detected by MTT method after co-cultured with different densities of MSC.

[Effects of quercetin on multidrug resistance and expression of related genes in human erythroleukemic K562/a cells].

The study was aimed to investigate the effect of quercetin, flavonoid molecules on reversing leukemia multidrug resistance and its mechanism. K562/A cells were cultured in vitro with different concentrations of quercetin. Cell growth inhibition and adriamycin (ADR) sensitivity were detected by MTT method.

[Apoptosis-inducing effect of quercetin and kaempferol on human HL-60 cells and its mechanism].

The purpose of this study was to explore the anti-leukemia effect of quercetin and kaempferol and its mechanism. The HL-60 cells were used as the leukemia models. The inhibitory effects of quercetin and kaempferol on growth of HL-60 cells was assessed by MTT assay.

Recombinant human granulocyte colony-stimulating factor significantly decreases the expression of CXCR3 and CCR6 on T cells and preferentially induces T helper cells to a T helper 17 phenotype in peripheral blood harvests.

The aim of this study was to investigate the expression of chemokine receptors on T cells and functional changes of T helper (Th) cells in peripheral blood stem cell (PBSC) harvests after treating healthy donors with recombinant human granulocyte colony-stimulating factor (rhG-CSF). Using multiparameter flow cytometry, we analyzed the expression of CXCR3 and CCR6 on T cells and the production of interferon-gamma (IFN-gamma), interleukin-4 (IL-4), and IL-17 by CD4(+) Th cells in PBSC grafts of healthy donors after in vivo rhG-CSF application. Alterations in the relative expression levels of T cell receptor beta variable (TCRBV) family members were determined using real-time polymerase chain reaction (PCR).

[Correlation of chemokine CCL-2/MCP-1 level in the plasma with aGVHD and idiophathic pneumonia syndrome after allogeneic hematopoietic stem cell transplantation].

The aim of this study was to investigate the relationship between the plasma levels of chemokine CCL-2/MCP-1 and acute graft-versus-host disease (aGVHD) and/or idiopathic pneumonia syndrome (IPS) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). ELISA assays were used to detect the plasma level of CCL-2/MCP-1 of 22 patients who received allo-HSCT, including 14 patients without or with grade I, 8 patients with grade II - IV aGVHD, respectively. 8 out of 22 patients were also diagnosed with IPS clinically.

[Dendritic cells elicit cellular immune response by targeting to capture breast cancer cells].

Objective: To investigate the specific anti-breast cancer immune response induced by dendritic cells (DC) loaded with trastuzumab and apoptotic Her-2+ breast cancer cells.

Methods: DCs were generated from healthy peripheral blood mononuclear cells (PBMCs) in the presence of recombinant cytokines GM-CSF, IL-4 and TNF-alpha. Mature DCs were harvested after 7 days' co-culture of PBMCs and trastuzumab-treated apoptotic SKBr3 cells.

[Human/mouse chimeric anti-CD20 monoclonal antibody enhances antigen presentation in dendritic cells and induces anti-lymphoma CTL effects].

In order to investigate the cellular immunoresponses mediated by chimeric anti-CD20 monoclonal antibody (anti-CD20 McAb) through dendritic cells (DCs), mononuclear cells were isolated from human peripheral blood (PBMNC) and DCs from PBMNCs in vitro were generated with normal methods. Human CD20 positive lymphoma cell line-Raji cells were treated with different methods including treatment with anti-CD20 McAb (group R), treatment with heat-induced apoptosis (group A) and treatment with anti-CD20 McAb+heat-induced apoptosis (group R+A), then Raji cells treated with above-mentioned methods as tumor antigen were loaded on DCs. The phagocytosis of DCs to tumor antigen was observed by transmission electron microscope (TEM), the auto-mixed lymphocyte reaction was performed with antigen-primed DCs, the release of INF-gammafrom effector cells was detected by ELISPOT, the killing of effector cells on Raji cells was assayed by MTT.

[rhIL-11 accelerates the engraftment of platelets after unrelated cord blood transplantation].

Objective: To observe whether rhIL-11 could accelerate the engraftment of platelets after unrelated cord blood transplantation (CBT).

Methods: Nine patients (3 children and 6 adults) were enrolled in this study. The degree of HLA disparity was 0-2 loci.

[In vitro study about the inhibitory effect of CDAII in combination with sodium butyrate on breast cancer cells].

Objective: To investigate if the combined use of CDAII and sodium butyrate can induce demethylation and re-expression of retinoic acid receptor beta2 (RARbeta2) gene in cultured human breast cancer cells MCF7. To explore if the two drugs can inhibit cell growth and induce cell apoptosis synergetically.

Methods: MCF7 cell line was treated with CDAII, sodium butyrate, combination of the two drugs respectively.

[Effect of the new human transcription factor hBKLF on the proliferation, differentiation of K562 cell line and hemoglobin synthesis].

The human basic Krüppel-like factor (hBKLF) is a newly cloned human transcription factor from the cDNA library of fetal liver. It belongs to the Krüppel-like transcription factor family. Previous expression study showed that it is a hematopoietic related factor.

[Immunological effector cells enhance apoptosis induced by adriamycin in a multi-drug resistant human breast cancer cell line].

Objective: To investigate the effects of immunologic effector cells to enhance apoptosis induced by adriamycin (ADR) in multi-drug resistant human breast cancer cell line MCF7/ADR.

Methods: The immunologic effector cells were induced and expanded by IFN-gamma, McAb CD3, IL-1 and IL-2. The expression of P-glycoprotein (P-gp) and its relation to apoptosis in target cells were detected by TUNEL technique and immunohistochemical staining.

[Influence of HSP70 on combined method of hyperthermia and immunologic effector cells to treat cancer].

Objective: To investigate hyperthermia enhanced expression of heat shock protein70 (HSP70) in breast cancer drug sensitive cell line MCF7 and multi-drug resistant (MDR) cell line MCF7/ADR, so as to increase cytotoxic activity of immunologic effector cells against the target cells, and to provide an experimental basis for cell immunotherapy based on hyperthermia.

Methods: The immunological effector cells were induced and expanded by cytokines. Breast cancer cell lines MCF7 and MCF7/ADR were treated at 42 degrees C for an hour.

[Construction of recombinant vector expressing ALAS2 gene in X-linked sideroblastic anemia].

X-linked sideroblastic anemia (XLSA) is caused by mutations of erythroid-specific 5-aminolevulinate synthetase (ALAS2) gene. In this study a eukaryotic expression vector of ALAS2 was constructed and transfected into eukaryotic cells to observe the expression of ALAS2 gene. The full length cDNA of ALAS2 gene was inserted into plasmid pDs-red2-N1, named pDs-red2-N1/ALAS2.

Cytokine-induced killer cells showing multidrug resistance and remaining cytotoxic activity to tumor cells after transfected with mdr1 cDNA.

Background: Routine treatment of cancer such as surgery, radiation or chemotherapy is sometimes unable to eradicate metastatic malignant cells. So we tried a new method and increased the adoptive immunotherapy of Cytokine-induced killer (CIK) cells in tumor patients and the multidrug resistance (mdr1) cDNA was transfected into CIK cells.

Methods: CIK cells were obtained from peripheral blood and induced by IFN-gamma, anti-CD3 monoclonal antibody, IL-2 and IL-1.

[Re-expression of p16 gene in myeloma cell line U266 by arsenic trioxide].

Background & Objective: DNA methylation status regulates gene expression and is associated with oncogenesis. Demethylation of DNA has been proposed as a possible new strategy for cancer prophylaxis and treatment. S-adenosylmethionine is required as methyl donor for both arsenic metabolism and DNA methylation.

[CIK cells acquired multidrug resistance and maintained cytotoxic activity to tumor cells after mdr1 gene transfection].

Objective: To investigate whether the cytokine-induced killer (CIK) cells could acquire multidrug resistance and maintain the original cytotoxic activity after multidrug resistance (mdr1) genes transfection.

Methods: CIK cells were generated from peripheral blood cultured with IFN-gamma, CD(3) monoclonal antibody, IL-2, IL-1 and transfected with a plasmid (pHamdr) containing human mdr1 gene via electroporation. RT-PCR method was used to assay mRNA expression of mdr1 gene in transfected CIK cells, flow cytometry with anti-P-gp monoclonal antibody to detect P-glycoprotein (P-gp) expression on CIK cells membrane, and MTT assay to compare both the multidrug resistance to doxorubicin and colchicines and cytotoxic activity to human mammary cancer cell line MCF7 between transfected and non-transfected CIK cells.

[Reversal effect of berbamine on multidrug resistance of K562/A02 cells and its mechanism].

This preject is to explore the reversal efficacy of calmodulin antagonist berbamine (BBM) on multidrug resistance (MDR) and its mechanism. Human erythroleukemic cell line K562 and its adriamycin-resistant counterpart K562/A02 were used in the study. The cells were co-cultured with ADR and BBM in different concentrations.

[The experimental study on the idiotypic nucleic Acid vaccine constructed from the genomic DNA to lymphoma].

This study was to investigate the anti-lymphocytic malignancy immunologic effects induced by two types of the idiotypic nucleic acid vaccines which were constructed from the genomic DNA and RNA of the human B lymphoma cell line respectively. Namalwa cell line and BALB/c mice were used as the models. The gene fragments of the IgH variable region (IgHV), which were obtained from the genomic DNA and RNA of Namalwa cell respectively, were cloned into the eukaryocytic expression vector pcDNA 3.