Acute Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; prevention & control ; therapy ; Postoperative Period ; Secondary Prevention

Hematopoietic Stem Cell Transplantation ; Humans ; Transplantation, Homologous ; Treatment Outcome

Dideoxynucleosides ; False Positive Reactions ; Fluorine Radioisotopes ; Fluorodeoxyglucose F18 ; Humans ; Inflammation ; diagnosis ; Neoplasm Staging ; Neoplasms ; diagnosis ; metabolism ; pathology ; radiotherapy ; Positron-Emission Tomography ; methods ; Radiotherapy, Intensity-Modulated ; Sensitivity and Specificity ; Tomography, X-Ray Computed ; Treatment Outcome

Sphingosine 1-phosphate (S1P) is an important biologically active lysophospholipid that transmits signals through a family of G-protein-coupled receptors (GPCRs) to regulate the vital functions of several types of immune cells. The S1P GPCRs suppress both generation of specialized functional cytokines, such as IFN-gamma and IL-4, and proliferation of T-cells. Although S1P is chemotactic to T cells, B cells, dendritic cells, and natural killer cells, the major effect of S1P on the immune system is the regulation of lymphocyte recirculation and tissue distribution by S1P and S1P1. Chemotactic response of CD4+CD25+ regulatory T cells to S1P is reduced, but its optimal suppressive activities require S1P. FTY720, a new class of immunomodulator, is rapidly phosphorylated by sphingosine kinase 2 in vivo to form the biologically active phosphorylated-FTY720 (FTY720-P), which closely resembles S1P. The FTY720-P is a true agonist for S1P1, S1P3, S1P4, and S1P5, it affects the tissue distribution and functional activity of T cells, B cells, dendritic cells and regulatory T cells. FTY720 were demonstrated to be a hypotoxic, great effective and reversible immunosuppressive efficacy to prevent allograft rejection and treat some autoimmune diseases. In this article, the synthesis and metabolism of S1P, the expression of S1P GPCRs in immune cells, the effect of S1P on immune cells, the drugs targeted to S1P GPCRs and their clinical implications are reviewed.
Fingolimod Hydrochloride ; Humans ; Immunomodulation ; physiology ; Lysophospholipids ; immunology ; physiology ; Propylene Glycols ; metabolism ; Receptors, G-Protein-Coupled ; immunology ; physiology ; Sphingosine ; analogs & derivatives ; immunology ; metabolism ; physiology

This study was aimed to clone human intercellular adhesion molecule-1 (ICAM-1) gene, to transfect the constructed eukaryotic expression vector ICAM-1-GFP into CHO cells, as well as to detect ICAM-1-GFP expression in CHO cells binding with Molt-4 cells. ICAM-1 cDNA gene was amplified by RT-PCR and inserted in PMD(18)-T vector. Then ICAM-1 cDNA from pMD18-ICAM-1 vector was subcloned into eukaryotic expression vector pEGFP-C1 to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing were used to confirm the recombinant vector. After stable transfection of CHO-K1 cells with the recombinant vector, the expression and subcellular localization of ICAM-1-GFP were detected by RT-PCR, flow cytometry and fluorescence microscopy. The function of ICAM-1-GFP fusion protein was assessed by the binding of ICAM-1-GFP/CHO cells to Molt-4 cells. The results showed that 1622 bp full-length ICAM-1 cDNA obtained and was successfully ligated with pMD(18)-T-vector, subcloned to construct recombinant ICAM-1-pEGFP-C1 vector. Restriction analysis and DNA sequencing indicated that recombinant ICAM-1-GFP was successfully constructed and ICAM-1-GFP was expressed stably in CHO cells. ICAM-1-GFP expression was only observed in the cytoplasm of ICAM-1-GFP/CHO cells by fluorescence microscopy. The ICAM-1-GFP/CHO cells were bound to PMA-treated Molt-4 cells. The expression of MEM-148 was very weak in PMA-treated Molt-4 cells. It is concluded that the ICAM-1-GFP eukaryotic expression vector has been constructed successfully and expresses stably in CHO cells. PMA can increase the binding of Molt-4 cells to ICAM-1-GFP/CHO cells by inducing specialized form of ICAM-1 clustering.
Animals ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; Recombinant Proteins ; genetics ; Transfection

As a functionally and phenotypically distinctive T cell subpopulation, CD4+CD25+ regulatory T cells are anergic and retain their ability to suppress antigen-driven response of CD4+CD25- cells in a contact-dependent manner or through a way of secreting immunosuppressive cytokines such as IL-10 and TGF-beta. Graft-versus-host disease (GVHD) is one of the major complications following allogeneic hematopoietic stem cell transplantation (HSCT). Recently, some researches on the relationship between donor CD4+CD25+ regulatory T cells and GVHD severity produced two contradictory conclusions: one is CD4+CD25+ regulatory T cells that can prevent GVHD efficiently; the other is that GVHD is associated with the increased numbers of peripheral blood CD4+CD25+ regulatory T cells. The answer to this question will provide a new idea for clinic therapy of GVHD. In this review some new research progresses in the related area, such as the CD4+CD25+ regulatory T cells, the phenotype, characteristics, immunoregulatory mechanisms of CD4+CD25+ regulatory T cells, as well as the relation of CD4+CD25+ with GVHD were presented.
CD4 Antigens ; analysis ; Graft vs Host Disease ; etiology ; immunology ; Humans ; Interleukin-2 Receptor alpha Subunit ; analysis ; T-Lymphocytes, Regulatory ; cytology ; immunology ; metabolism

This study was aimed to investigate the expression level of stromal cell derived factor-1 gene (SDF-1) in bone marrow mesenchymal stem cells (MSC) of patients with myelodysplastic syndrome (MDS). The MSC from bone marrow samples of MDS patients were isolated, cultured and expanded, the morphology and immunophenotype of MSC were analyzed. The expression levels of SDF-1 and internal reference GAPDH in MSC of MDS patients were detected by real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) method and were compared with expression levels of healthy donors. The results showed that the expression levels of SDF-1 in MDS patients were significantly different from those in healthy donors (1.53 +/- 0.92 vs 5.51 +/- 0.99) (P < 0.01). SDF-1 gene expression levels in bone marrow MSC of MDS patients were significantly higher than that in MSC derived from healthy donors. It is concluded that the abnormal expression of SDF-1 gene in MSC may influence the regulation of hematopoiesis of the bone marrow microenvironment in MDS patients and it is worthy of further investigation for new clue on etiological mechanism and treatment of MDS.
Adult ; Aged ; Bone Marrow Cells ; metabolism ; pathology ; Cells, Cultured ; Chemokine CXCL12 ; biosynthesis ; genetics ; Female ; Humans ; Male ; Mesenchymal Stromal Cells ; metabolism ; pathology ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; metabolism ; pathology

OBJECTIVETo explore a new method for in vitro expansion of human mesenchymal stem cells (MSC) by using platelet-rich plasma (PRP) in place of fetal calf serum (FCS).

METHODSBone marrow(BM) samples were obtained from the proximal femurs of patients with normal haematopoietic function undergoing total hip arthroplasty. 2 x 10(5)/cm2 BM nucleated cells were seeded in 25 cm2 flasks for MSC cultivation containing one of the 3 mediums: complete Dexter medium with 12.5% FBS and 12.5% horse serum (medium1), alpha-MEM with 10% FCS (medium2) and alpha-MEM with 5% PRP(medium3). At the same time, 1 x 10(6) nucleated BM cells and same amount of nucleated cells from iliac aspirate were seeded in 25 cm2 tissue flasks, colony forming unit-fibroblast (CFU-F) assay.

RESULTSCulture-expanded cells from proximal femurs presented a typical fibroblast-like morphology. Cells were positive for SH2 (CD 105), SH3 (CD73), Thy-1 (CD90), while negative for CD34 and CD45. On induction, these cells could differentiate into osteoblasts. A significantly higher proliferative capacity of MSCs expanded in medium3 was observed in comparison to those in mediuml or 2 without alteration of the phenotype and the differentiation property. CFU-F assays indicated that bone marrow from the proximal femoral contained significantly more CFU-F than that from iliac aspirate.

CONCLUSIONPlatelet-rich plasma can be used in place of FCS to provide a safer and more effective culture condition to expand MSCs for clinical purpose. The proximal femur BM cells can be obtained in hip surgeries.

The interaction of killer cell immunoglobin-like receptors (KIR) with HLA molecules has particular relevance to the genetics, immune responses and allogeneic stem cell transplantation. The genes of KIR and HLA are located in different chromosomes and segregate independently. The repertoire of KIR molecules varies among NK cells and is determined by the KIR genotype. The HLA genotype has only subtle impact on the KIR phenotype. Three major HLA specificity groups are recognized by KIRs. Donor versus recipient NK-cell alloreactivity, when recipients lack HLA ligand for their donor inhibitory KIR, can benefit allogeneic stem cell transplantation, especially the HLA haploidentical hematopoietic stem cell transplantation. The outcome of stem cell transplantation can be best predicted by the presence of KIRs on the donor's NK cells and the absence of corresponding KIR ligand in the recipient's HLA repertoire-a receptor-ligand model. In this paper the interaction of KIR and HLA in hematopoietic stem transplantation, the genetic basis of KIR and HLA, the relation of KIR expression on NK cells with HLA and the role of KIR and HLA in immune responses were reviewed.
Hematopoietic Stem Cell Transplantation ; Histocompatibility Antigens Class I ; genetics ; immunology ; Humans ; Killer Cells, Natural ; cytology ; immunology ; Receptors, Immunologic ; genetics ; immunology ; Receptors, KIR ; Transplantation Immunology

Spectral karyotyping (SKY) is a novel cytogenetic technique, has been developed to unambiguously display and identify all 24 human chromosomes at one time without a priori knowledge of any abnormalities involved. SKY discerns the aberrations that can not be detected very well by conventional banding technique and fluorescent in situ hybridization (FISH). So SKY is hyper-accurate, hypersensitive, and hyper-intuitional. In this paper the basic principle of SKY technique and its application in leukemia cytogenetics were reviewed.
Humans ; Karyotyping ; Leukemia ; genetics ; pathology ; Spectral Karyotyping