In order to investigate the levels of stem cell factor (SCF) and its receptor c-kit protein and mRNA in pediatric aplastic anemia (AA) and their relevance to the pathogenesis, immunocytochemical and in situ hybridization were utilized to detect the expression of SCF and its receptor c-kit gene protein and mRNA, respectively in 59 children with AA and 51 normal controls. The relationship between SCF and c-kit and the pathogenesis of AA was analyzed subsequently. The results showed that the positive rate of SCF protein and mRNA expression in children with AA was significantly lower than that in healthy controls (P < 0.05). However, there was no significant difference in the positive rate of c-kit protein and mRNA expression between children with AA and control group (P > 0.05). It was concluded that the expression of SCF is significantly decreased in children with AA, which may be closely associated with the pathogenesis of the AA. c-kit may be unrelated to the development of pediatric AA. Therefore, AA in children may have abnormalities at SCF/c-kit signal transduction levels.
Anemia, Aplastic/etiology ; Anemia, Aplastic/*metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Receptors, Colony-Stimulating Factor/*biosynthesis ; Receptors, Colony-Stimulating Factor/genetics ; Stem Cell Factor/*biosynthesis ; Stem Cell Factor/genetics

OBJECTIVETo investigate the expression of the granulocyte-macrophage colony-stimulating factor (GM-CSF) and GM-CSF/IL-3/IL-5 receptor common beta chain (beta c receptor) in an adult patient with idiopathic pulmonary alveolar proteinosis (PAP), so as to demonstrate the possible association of the GM-CSF and beta c receptor with the pathogenesis of human PAP.

METHODSThe GM-CSF levels were measured with a commercial ELISA kit (sensitivity 5 pg/ml) and the beta c receptor expression on the cell surface was detected by flow cytometry analysis. Reverse transcription polymerase chain reaction (RT-PCR) analysis was employed to detect the expression of the GM-CSF mRNA and the beta c receptor mRNA in peripheral blood mononuclear cells and alveolar macrophages. The entire coding regions of the GM-CSF cDNA and the beta c receptor cDNA were sequenced by the Sanger dideoxy-mediated chain termination method to detect possible mutations.

RESULTSThe patient with PAP failed to release the GM-CSF protein either from circulating mononuclear cells or from alveolar macrophages. The expression of the GM-CSF mRNA was normal after the stimulation of lipopolysaccharide, whereas a point mutation at position 382 of the GM-CSF cDNA from "T" to "C" was revealed by cDNA sequencing, which caused a change in amino acid 117 of the protein from isoleucine to threonine. The beta c receptor expression on the cell surface was normal, and the beta c receptor mRNA expression and the sequence of the entire coding region of the beta c receptor were also normal.

CONCLUSIONSThe decreased GM-CSF production is associated with the pathogenesis of human PAP. A point mutation of the GM-CSF cDNA may contribute to the decreased GM-CSF production in our adult PAP patient. The mutation of the beta c receptor in some of paediatric patients with PAP may not be a common problem in adult patients.

DNA, Complementary ; chemistry ; Granulocyte-Macrophage Colony-Stimulating Factor ; analysis ; biosynthesis ; Humans ; Male ; Middle Aged ; Pulmonary Alveolar Proteinosis ; etiology ; metabolism ; RNA, Messenger ; analysis ; Receptors, Cytokine ; biosynthesis ; genetics

The aim of this research was to understand the influence of rhG-CSF on the sphingosine kinase (SphK) activity of monocytes. The peripheral blood monocytes were collected from 6 peripheral blood progenitor cell donors on the fifth day of mobilization with rhG-CSF and from 5 blood donors' buffy coats. The mRNA expressions of monocyte G-CSF receptor and SphK were tested with RT-PCR. The changes of SphK activity of monocytes were assayed after being treated with rhG-CSF. The results showed that the two kinds monocytes collected from both blood donors and peripheral blood progenitor cell donors mobilized with rhG-CSF expressed mRNA of G-CSF receptor and SphK. The SphK activity of monocytes collected from blood donors was not changed significantly after being treated with rhG-CSF (P > 0.05). The SphK activity of monocytes collected from peripheral blood progenitor cell donors transiently increased by (39.6 - 87.2)% after being treated by means of rhG-CSF (P < 0.05) without obviously dose-dependent effect. It is concluded that the SphK activity of monocytes collected from peripheral blood progenitor cell donors can be activated by rhG-CSF.
Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; Humans ; Monocytes ; cytology ; enzymology ; Phosphotransferases (Alcohol Group Acceptor) ; drug effects ; metabolism ; Receptors, Granulocyte Colony-Stimulating Factor ; biosynthesis ; genetics ; Recombinant Proteins

To evaluate soluble GM-CSF-Ralpha expression in patients with acute myeloid leukemia (AML) and its clinic significance, plasma concentration of solGM-Ralpha in de novo 66 patients with AML was detected by enzyme-linked immuno-sorbent assay, and the relationship between solGM-Ralpha levels and various clinical parameters was analyzed. The result showed that the levels of solGM-Ralpha in plasma of patients with AML were significantly higher than that in plasma of normal controls; the lowest level of solGM-Ralpha was found in plasma of patients with AML-M3 (3897.75 +/- 2651.43 pg/ml), the highest level of solGM-Ralpha was observed in plasma of patients with AML-M5 (9990.92 +/- 6325.43 pg/ml). Patients with high level of solGM-Ralpha were generally accompanied with a distinct clinical picture, including higher counts of white blood cell and myeloid precursors, as well as higher expression of CD34, CD95 and CD116 antigen. It is concluded that the high level of solGM-Ralpha in plasma of patients may suggest AML poor prognosis and play a role in pathogenesis of leukemia, the GM-CSF and its receptor solGM-Ralpha needs further study.
Adolescent ; Adult ; Aged ; Antigens, CD34 ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Leukemia, Myeloid, Acute ; blood ; Male ; Middle Aged ; Prognosis ; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor ; biosynthesis ; blood ; fas Receptor ; blood

OBJECTIVETo study the expression and significance of granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors in human prostate cancer.

METHODSSP immunohistochemical method was employed to investigate the expression of alpha subunit of GM-CSF receptors in 48 cases of primary prostate cancer, 20 benign prostate hyperplasia samples and four kinds of cancer cell lines K562, PC-3M, HL-60 and MCF-7.

RESULTSThe total positive percentage of GM-CSF expression in prostate cancer was 79.2%. The positive percentages in the groups with Gleason score 2-4, 5-8, and 9-10 were 75%, 82.3% and 81.2% respectively. The four kinds of cancer cell lines had prominent GM-CSF receptor alpha subunit expression.

CONCLUSIONIt suggests that both hyperplastic and neoplastic prostate tissues are responsive to GM-CSF and the extensive expression of GM-CSF receptors is an important characteristic of prostate cancer.

Animals ; Blotting, Western ; Cell Line, Tumor ; HL-60 Cells ; Humans ; Immunohistochemistry ; K562 Cells ; Male ; Prostatic Hyperplasia ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; Rabbits ; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor ; biosynthesis

To explore the dynamic change of CD34(+) cell expressing adhesion molecules in bone marrow and peripheral blood during mobilization with combination of chemotherapy and G-CSF and its clinical significance, mononuclear cells of bone marrow and peripheral blood from malignant hematopathy cases before and after mobilization with G-CSF were labeled by CD45-CY-Chrome, PE conjugated anti-CD34, and FITC conjugated anti-CD44, anti-CD49d, anti-CD62L and anti-CXCR4. For three-color fluorescence analysis by flow cytometry was performed on a FACScalibur. Also the relationship between the number of subpopulations in different expressions of adhesion molecules infused and the time of recovery in different blood cells after transplantation was evaluated. Results showed that a significantly lower expression of CD44(+) and CD49d(+) on CD34(+) cells in bone marrow after mobilization compared to that before mobilization, whereas great higher expression of CD44(+), CD49d(+), anti-CD62L(+) and lower of anti-CXCR4(+) in peripheral blood were observed after mobilization. No significant relations were found between expression of different adhesion molecules on CD34(+) cells infused and the time of reconstitution in blood cells after transplantation. It was concluded that this mobilizing regimen could downregulate the expressions of CD44, CD49d, CD62L, and anti-CXCR4 on CD34(+) cells in bone marrow, it may related to mobilization of CD34(+) cells from marrow to blood, and homing of blood CD34(+) cells into marrow.
Adolescent ; Adult ; Antigens, CD34 ; immunology ; Bone Marrow Cells ; immunology ; metabolism ; Cell Adhesion Molecules ; biosynthesis ; blood ; Female ; Flow Cytometry ; methods ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematologic Neoplasms ; immunology ; metabolism ; therapy ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cell Transplantation ; Hodgkin Disease ; immunology ; metabolism ; therapy ; Humans ; Hyaluronan Receptors ; biosynthesis ; Integrin alpha4 ; biosynthesis ; L-Selectin ; biosynthesis ; Leukocytes, Mononuclear ; immunology ; metabolism ; Lymphoma, Non-Hodgkin ; immunology ; metabolism ; therapy ; Male ; Middle Aged ; Multiple Myeloma ; immunology ; metabolism ; therapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; metabolism ; therapy

To investigate the effect of in vivo rhG-CSF on the CXCR-4 expression of hematopoietic progenitor or stem cells in bone marrow and peripheral blood, the expressions of CXCR-4 on CD34(+) cells and mononuclear cells of bone marrow and peripheral blood from healthy donor before and after mobilization were detected by three-color fluorescence analysis. The results showed that a significantly higher expression of CXCR4 on CD34(+) cells of bone marrow and mononuclear cells of peripheral blood, as compared to those before mobilization. There were no significant differences of CXCR-4 expression of CD34(+) cells in peripheral blood after mobilization, as compared with steady state bone marrow, and no dynamic change of mononuclear cells expressing CXCR-4 in bone marrow before and after mobilization. Significant positive correlation were found between the percentage of CD34(+) cells in bone marrow before mobilization and that in bone marrow and peripheral blood after mobilization; furthermore, the percentage of CD34(+) cells of bone marrow before mobilization had a positive correlation with both the count of CD34(+) cells per kilogram on the day of collection in bone marrow and peripheral blood after mobilization. It is concluded that the mobilization of hematopoietic cells may be involved in the signaling of SDF-1/CXCR-4 according to the increase of the surface expression of CXCR-4 on CD34(+) cells in bone marrow and on the MNC in peripheral blood after mobilization; meanwhile, the high surface expression of CXCR-4 may contribute to the MNC engraftment, monitoring the percentage of CD34(+) cells in bone marrow before mobilization can be regarded as a predictive factor for mobilization outcome.
Adolescent ; Adult ; Aged ; Antigens, CD34 ; blood ; Blood Donors ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Female ; Flow Cytometry ; methods ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; cytology ; drug effects ; metabolism ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Receptors, CXCR4 ; biosynthesis ; blood ; Recombinant Proteins