Upregulated TCRζ enhances interleukin-2 production in T-cells from patients with CML.

T-cell immunodeficiency is a common feature in patients with chronic myeloid leukemia (CML), and deficiency in CD3 levels was detected in T cells from these patients, which may represent a characteristic that is related to a lower T cell activation. In this study, we explored the possibility that forced TCRζ gene expression may upreg-u-late T cell receptor (TCR) signaling activation and reverse interleukin-2 (IL-2) production in T cells from patients with CML. A recombinant eukaryotic vector expressing TCRζ was transfected into T cells by nucleofection.

Alteration of the gene expression profile of T-cell receptor αβ-modified T-cells with diffuse large B-cell lymphoma specificity.

Antigen-specific, T-cell receptor (TCR)-modified cytotoxic T lymphocytes (CTLs) that target tumors are an attractive strategy for specific adoptive immunotherapy. Little is known about whether there are any alterations in the gene expression profile after TCR gene transduction in T cells. We constructed TCR gene-redirected CTLs with specificity for diffuse large B-cell lymphoma (DLBCL)-associated antigens to elucidate the gene expression profiles of TCR gene-redirected T-cells, and we further analyzed the gene expression profile pattern of these redirected T-cells by Affymetrix microarrays.

[Supportive effects of conditioned culture media of human umbilical cord mesenchymal stem cells on hematopoiesis in vitro].

This study was aimed to explore whether the conditioned culture medium of human umbilical cord-derived mesenchymal stem cells (hUC-MSC) has supportive effects on hematopoiesis in vitro. hUC-MSC were cultured in 75 cm(2) culture flasks at a concentration of 2×10(6) cells per flask. After 48 h, the conditioned culture medium was harvested.

Comparison of the distribution and clonal expansion features of the T-cell γδ repertoire in myelodysplastic syndrome-RAEB and RAEB with progression to AML.

Myelodysplastic syndrome (MDS) is a heterogeneous group of clonal hematopoietic stem cell diseases. Approximately 30% of patients with MDS will develop acute myeloid leukemia (AML). Immune dysregulation may contribute to MDS initiation and progression.

[Expression of reconstructed BCR-ABL-pIRES-SEA plasmids in the skeletal muscles of BALB/c mice].

This paper is aimed to investigate the transcription and expression of BCR-ABL-pIRES-SEA fusion gene vaccines in vivo in mice. The reconstructed plasmids (BCR-ABL-pIRES-SEA) which were developed previously in our laboratory were injected into the skeletal muscles of BALB/c mice at 14d intervals for three cycles. The transcription and expression of BCR-ABL and staphylococcal enterotoxin A (SEA) in injection site were detected using RT-PCR and immunohistological methods.

Expression feature of CD3, FcεRIγ, and Zap-70 in patients with chronic lymphocytic leukemia.

In leukemia patients, T-cell function has been suppressed with the disease progress. Patients with chronic lymphocytic leukemia (CLL) are all to a degree immunodeficient. In order to elucidate the feature of T-cell receptor signal transduction in CLL, the expression levels of CD3γ, δ, ε, and ζ chain, FcεRIγ, and Zap-70 genes in peripheral blood mononuclear cells (PBMCs) were analyzed.

Mutations increased overexpression of Notch1 in T-cell acute lymphoblastic leukemia.

Background: The Notch signaling pathway is crucial in T-cell development, Notch1 mutations are frequently present in T-cell acute lymphoblastic leukemia (T-ALL). To investigate the feature of Notch1 mutation and its corresponding expression level in Chinese patients with T-ALL, detection of mutation and the expression level of Notch1 gene was preformed using RT-PCR, sequencing and real-time PCR respectively.

Results: Two Notch1 point mutations (V1578E and L1593P) located on HD-N domain were identified in three cases out of 13 T-ALL patients.

The role of BCL11B in regulating the proliferation of human naive T cells.

The effect of the B-cell chronic lymphocytic leukemia/lymphoma 11B gene (BCL11B) on human T-cell regulation remains unclear. To characterize the functions of BCL11B, recombinant BCL11B and BCL11B siRNA were transfected into human naive T cells to overexpress or knock down BCL11B expression, respectively. After BCL11B overexpression, the proliferation ability and the T-helper (Th) subset were increased, whereas no significant alteration in the expression pattern and clonality of the T-cell receptor Vβ subfamilies was observed.

Altered expression of the TCR signaling related genes CD3 and FcεRIγ in patients with aplastic anemia.

Background: Aplastic anemia (AA) is characterized by pancytopenia and bone marrow hypoplasia, which results from immune-mediated hematopoiesis suppression. Understanding the pathophysiology of the immune system, particularly T cells immunity, has led to improved AA treatment over the past decades. However, primary and secondary failure after immunosuppressive therapy is frequent.

[Effect of superantigen SEA on mitochondrial membrane potential of Molt-4 cell].

Aim: To explore the effect of superantigen staphylococcal enterotoxin A(SEA) on mitochondrial membrane potential of Molt-4 cell.

Methods: Cell counting kit-8(CCK-8) was used to detect the proliferation of T cell in different concentration and time, We employed JC-1 to estimate mitochondrialmembrane potential (δψm) of Molt-4 cell induced by SEA using flow cytometry (FCM).

Results: The proliferative activity of T cell treated with SEA(1 mg/L) was changed obviously compared with control.

A change in CD3γ, CD3δ, CD3ϵ, and CD3ζ gene expression in T-lymphocytes from benzene-exposed and benzene-poisoned workers.

Benzene is known to be highly toxic to a variety of cell types, including lymphocytes. A previous study showed that T-lymphocyte immune function disorder might be related to benzene exposure. To elucidate characteristics of TCR signal transduction in benzene-exposed workers, expression levels of CD3γ, CD3δ, CD3ϵ, and CD3ζ genes in peripheral blood mononuclear cells (PBMC) were analyzed.

[Change in the pattern of peripheral blood TCR V β gene repertoire in occupational lead-exposed workers].

Aim: To investigate the distribution and clonality of TCR Vβ subfamily in peripheral blood from workers exposed to lead, in order to understand the change in T cell immunity of occupational lead-exposed workers.

Methods: The CDR3 of TCR Vβ 24 subfamily genes was amplified in peripheral blood mononuclear cells (PBMC) from 6 lead-exposed workers and 6 healthy individuals using RT-PCR, the positive PCR products were further subjected by genescan analysis to identify T cell clonality.

Results: All 24 TCR Vβ subfamilies were detected in PBMC from 6 healthy individuals which all have polyclonal patterns.

[Establishment of stable subline of K562 cells expressing human leucocyte antigen a1101].

The aim of this study was to establish a stable subline of K562 cells expressing the HLA-A(*)1101 protein, which was expected to provide target cells for characterizing the HLA-I restrictive antigen specific cytotoxic T lymphocyte (CTL) effects against chronic myeloid leukemia (CML). The HLA-A(*)1101 protein encoding gene was amplified from peripheral blood mononuclear cell (PBMNC) of CML patient by RT-PCR; the 2A peptide linker (D-V-E-X-N-P-G-P) gene was linked to the 3'terminal of the HLA-A(*)1101 gene by recombinant PCR, then the recombinant was cloned into the pEGFP-N3 plasmid which contains an enhanced green fluorescent protein gene, and the eukaryotic recombinant expression vector containing HLA-A(*)1101-T2A-EGFP transcription box was constructed; the pEGFP-N3 vector and recombinant vector was separately electroporated into K562 cells. The expression of GFP was monitored by fluorescence microscopy, finally stably transfected sublines of K562 cells containing HLA-A(*)1101 gene, and of K562 containing pEGFP-N3 vector were obtained by G418 selection; the transcriptional or translational expression of HLA-A(*)1101 gene was detected with RT-PCR and flow cytometry respectively.

[Immune regulation role of A20 and its clinical significance].

A20 was originally identified as a TNFα-induced protein 3 (TNFAIP3), a key regulator of inflammation signalling pathways, as well as a NF-κB inhibitor. It plays a critical role in regulation of innate and adoptive immunity. Recently, A20 has also been proposed to function as a tumor suppressor.

Characterization of the CDR3 structure of the Vβ21 T cell clone in patients with P210(BCR-ABL)-positive chronic myeloid leukemia and B-cell acute lymphoblastic leukemia.

The clonally expanded T cells identified in most cancer patients that respond to tumor-associated antigen such as P210(BCR-ABL) protein have definite, specific antitumor cytotoxicity. T cell receptor (TCR) Vβ CDR3 repertoire diversity was analyzed in patients with chronic myeloid leukemia (CML) and BCR-ABL(+) B-cell acute lymphoblastic leukemia (B-ALL) by GeneScan. A high frequency of oligoclonal expansion of the TCR Vβ21 subfamily was observed in the peripheral blood of CML and B-ALL patients.

Down regulation of BCL11B expression inhibits proliferation and induces apoptosis in malignant T cells by BCL11B-935-siRNA.

To screen the highly efficient and specific B-cell chronic lymphocytic leukemia/lymphoma 11B (BCL11B) small interfering RNA (siRNA) which are able to downregulate the BCL11B gene expression in human T-cell acute lymphoblastic leukemia, thereby inhibiting the leukemic T-cell proliferation and inducing apoptosis, four BCL11B-siRNAs and the scrambled non-silencing siRNA control (sc) were designed and obtained by chemosynthesis. After nucleofection, BCL11B expression in the mRNA and the protein levels were measured by qRT-PCR and immunoblotting, respectively. The biological consequences based on the highly efficient and specific BCL11B-siRNA were demonstrated by CCK-8 kit, morphological changes (Hoechst 33258 staining), high-resolution imaging, and flow cytometry.

The expression pattern of CD3 chain genes in fetal/maternal interface.

In order to investigate the features of T-cell immune status in human placenta, the expression levels of CD3-gamma, -delta, -epsilon and -zeta chain genes in placenta were analyzed by real-time PCR. Umbilical cord blood obtained at delivery from the full-term healthy babies was used as a control. The beta2-microglobulin gene was employed as an endogenous reference, and the evaluations of mRNA expression level of each CD3 gene were used by the 2(-ΔC(t))×100% method.

Change in expression pattern of TCR-CD3 complex in patients with multiple myeloma.

In haematological malignancy, cell-mediated immunity has been shown to be suppressed in advanced disease. This immune dysfunction may be due, in part, to altered expression of the T cell receptor (TCR)-CD3 complex. The distribution and clonality of the TCR Vbeta repertoire and the expression levels of CD3gamma, CD3delta, CD3epsilon, and CD3zeta genes in T cells from patients with multiple myeloma (MM) were investigated.

[Abnormal increase in CD8(low) T lymphocyte in patients with occupational chronic lead poisoning].

Objective: To analyze the changes in CD8(low) T lymphocyte subsets in patients with occupational chronic lead poisoning.

Methods: Flow cytometric analysis was used to count the numbers of CD8+ cells. 23 patients with occupational chronic lead poisoning and 20 controls were examined.

[Expression of CD133 in the bone marrow of patients with myelodysplastic syndrome and its clinical significance].

Objective: To investigate the expression of CD133 in the bone marrow of patients with myelodysplastic syndrome (MDS) and explore its clinical significance.

Methods: The expression of CD133 and CD34/CD38 in the bone marrow was detected using flow cytometry in 31 cases of refractory anemia with excess blasts (RAEB), 10 cases of refractory cytopenia with multilineage dysplasia (RCMD) and 11 cases of aplastic anemia (AA).

Results: The percentage of CD133-expressing cells was 6.

Gene expression profiles in BCL11B-siRNA treated malignant T cells.

Background: Downregulation of the B-cell chronic lymphocytic leukemia (CLL)/lymphoma11B (BCL11B) gene by small interfering RNA (siRNA) leads to growth inhibition and apoptosis of the human T-cell acute lymphoblastic leukemia (T-ALL) cell line Molt-4. To further characterize the molecular mechanism, a global gene expression profile of BCL11B-siRNA -treated Molt-4 cells was established. The expression profiles of several genes were further validated in the BCL11B-siRNA -treated Molt-4 cells and primary T-ALL cells.

[Analysis of TCR gene rearrangement for identification of T cell leukemia clone by using fine-tiling aCGH].

Aim: To establish a new method which analyzes T cell receptor (TCR) gene rearrangement for identification of T cells acute lymphoblastic leukemia (T-ALL) clone, it will provide the basis for the study of T-ALL including the chromosome translocation involving TCR loci.

Methods: Total DNA was isolated from peripheral blood mononuclear cells (PBMC) of one case with T-ALL. Using the fine-tiling array comparative genomic hybridization (fine-tiling aCGH) to analyze the genomic DNA differences of the case and control group, we could find the breakpoints and their position in the chromosomes.

Expression and distribution of PPP2R5C gene in leukemia.

Background: Recently, we clarified at the molecular level novel chromosomal translocation t(14;14)(q11;q32) in a case of Sézary syndrome, which caused a rearrangement from TRAJ7 to the PPP2R5C gene. PPP2R5C is one of the regulatory B subunits of protein phosphatase 2A (PP2A). It plays a crucial role in cell proliferation, differentiation, and transformation.

Frequency analysis of TRBV subfamily sjTRECs to characterize T-cell reconstitution in acute leukemia patients after allogeneic hematopoietic stem cell transplantation.

Background: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) leads to a prolonged state of immunodeficiency and requires reconstitution of normal T-cell immunity. Signal joint T-cell receptor excision DNA circles (sjTRECs) are markers of developmental proximity to the thymus that have been used to evaluate thymic function related to T-cell immune reconstitution after HSCT. To assess the proliferative history in different T-cell receptor beta variable region (TRBV) subfamilies of T cells after HSCT, expansion of TRBV subfamily-naive T cells was determined by analysis of a series of TRBV-BD1 sjTRECs.

[TCRζ gene expression in TCR gene-modified T cells specific for DLBCL-associated antigen].

Aim: To analyze the expression level of TCRζ chain gene in the DLBCL-associated antigen-specific T cells before and after being activated by coculture with Toledo cells (DLBCL cell line).

Methods: Real-time PCR with SYBR GreenI technique was used for detecting TCRζ chain expression in activated and unactivated DLBCL-associated antigen-specific T cells. β2 microglobulin gene (β2M) was used as an endogenous reference.