Administration of G-CSF may not always respond in rise of neutrophil counts in different patient population. In order to understand a possible inter-relationship between the G-CSF and GM-CSF induced leukocyte responses and expression levels of receptors for G-CSF (G-CSFr) and GM-CSF (GM-CSFr), the levels of each receptor and CSF were measured in patients with basophilia (8), eosinophilia (14) and bacterial infection showing neutrophilia (12) in comparison with normal healthy adults (12) and children (14). G-CSFr was expressed in neutrophils in the largest amount followed by monocytes, but GM-CSFr was expressed more in monocytes than neutrophils. Lymphocytes and basophils did not express G-CSFr or GM-CSFr. The amount of GM-CSFr in neutrophils was present less in patients with infection than normal control (P = 0.031). The neutrophils expressed more G-CSFr than GM-CSFr. The quantity of G-CSFr in eosinophil showed marked interval change, higher in acute stage. The plasma concentrations of G-CSF in patients with infection were much higher than normal adults or children (117.95 +/- 181.16 pg/ml, P < 0.05). Binding assay with excess amount of CSFs could discriminate the patient who did not show any response to G-CSF or GM-CSF administration. After incubation with excess CSFs, more receptors were blocked in children than in adults (G-CSF P = 0.024, GM-CSF P = 0.006). These results indicate that the amount of CSFr in leukocyte varies in different types of leukocyte, and changes according to the patients' condition even in the same type of leukocyte, and the CSFrs of children bind to CSFs more than those of adults.
Adult ; *Bacterial Infections ; Basophils/chemistry ; Breast Neoplasms ; Child ; Colony-Stimulating Factors/*blood ; Eosinophilia ; Human ; Leukemia, Myeloid, Chronic ; *Leukocyte Disorders ; Monocytes/chemistry ; *Neoplasms ; Neutrophils/chemistry ; Receptors, Colony-Stimulating Factor/*analysis ; Receptors, Granulocyte Colony-Stimulating Factor/analysis ; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/analysis

To explore the reasonable procedures and strategies of diagnosis and treatment of congenital neutropenia (CN), clinical data and laboratory examination results of a boy suspected of CN were collected; gene ELA2, GFI1, HAX1, and WASp of whom were sequenced, granulocyte colony-stimulating factor receptor (G-CSFR) expression on neutrophil was analyzed, and cytoplasmic domain of G-CSFR was sequenced. The results showed that the diagnosis of non-syndromic variants of CN (NSVCN) was made on this patient according to the criteria; sequencing results revealed no mutation occurred in ELA2, GFI1, HAX1 and WASp; a normal expression level of G-CSFR on neutrophil from this patient was detected and no truncated mutation was found in the intracellular domain of G-CSFR. It is concluded that reasonable procedure of diagnosis and treatment of CN is established, and a sporadic NSVCN with no recognized pathogenic mutation is confirmed in this patient.
Child ; DNA Mutational Analysis ; Humans ; Male ; Neutropenia ; congenital ; diagnosis ; genetics ; therapy ; Receptors, Granulocyte Colony-Stimulating Factor ; metabolism

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

OBJECTIVE: To identify differentially expressed genes in peripheral blood mononuclear cells between patients with continuous mild-to-moderate asthma and healthy controls using mRNA microarray in order to explore the underlying signaling pathways and clarify the roles of CD4+ T cells in the pathogenesis of asthma. METHODS: Global transcriptomic profiles of the CD4+ T cells were defined by using Agilent Sure Print G3 Human GE 8×60K microarray. Enrichment pathways were analyzed with Ingenuity Pathway Analysis (IPA) software. RESULTS: Compared with controls, 805 genes were up-regulated, 192 were down-regulated in asthma patients. Among these, the expression of 38 annotated genes have varied by 4 times or more. Expression of CD300A was inversely proportional to the absolute value of eosinophils (r=-0.89, P=0.02) as well as the proportion of eosinophils (r=-0.94, P=0.004), while CSF1R was inversely proportional to PD20 (r=-0.83, P=0.04) and AQLQ (r=-0.88, P=0.02) by correlation analysis. CONCLUSION Numerous pathophysiological pathways may be involved in the pathogenesis of asthma. Above findings have provided a basis for the delineation the pathogenesis of asthma.
Antigens, CD ; genetics ; Asthma ; immunology ; CD4-Positive T-Lymphocytes ; cytology ; Case-Control Studies ; Eosinophils ; Gene Expression Profiling ; Humans ; Leukocytes, Mononuclear ; Oligonucleotide Array Sequence Analysis ; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Receptors, Immunologic ; genetics ; Transcriptome

In order to analyze the clinical characteristics and biological features of acute leukemia in elderly, 104 acute leukemia patients in elderly were retrospectively analyzed and compared with 71 acute leukemia patients below 60 years old. The results showed that: (1) the proportion of AML in the aged group (73%) was higher than that in the young group (54.9%), and the difference was statistically significant (P < 0.05), but AML (M3) was absent in the aged group; (2) the median of bone marrow blast cell in the aged group was significantly lower than that in the young group (P < 0.05); (3) in AML, the frequently of CD14 expression was higher in the aged group (18.8%) than that in the young group (2.6%), while the expression frequencies of CD15 (37.5%), CD117 (62.5%), and CD38 (59.4%) were respectively lower in the aged group than that in the young group which were (69.2%) for CD15, (89.7%) for CD17, and (84.6%) for CD38 respectively, and the difference was also statistically significant (P < 0.05). (4) CD19 was most frequently expressed in ALL of the aged group and the positive rate was 100%; (5) there was no significant difference in expression of special lineage antigens and overlapping lineage antigens between the aged group and the young group (P > 0.05); (6) the expression frequency of unfavorable karyotypes in the aged group was higher than that in the young group, and the difference was statistically very significant (P < 0.01); (7) the complete remission rate (CR rate) in the aged group was 42.9%, 2-year survival rate in the aged group was 5.4%, and treatment-related mortality rate in the aged group was 26.8%, while the CR rate in the young group was 76.6%, the difference was statistically significant (P < 0.05). It is concluded that the expression frequency of CD14 associated with unfavorable prognosis is higher in the aged group than that in the young group, while the expression frequency of CD15 associated with favorable prognosis is lower in the aged group than that in the young group. The expression frequency of unfavorable karyotypes in the aged group is higher than that in the young group. The CR rate of acute leukemia in elderly is low, thus the patients in elderly often have unfavorable prognosis.
Adult ; Age Factors ; Aged ; Aged, 80 and over ; Female ; Humans ; Immunophenotyping ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; immunology ; Lipopolysaccharide Receptors ; analysis ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; immunology ; Prognosis ; Receptor, Macrophage Colony-Stimulating Factor ; analysis ; Remission Induction ; Retrospective Studies

This study was performed to evaluate the effect of granulocyte-colony stimulating factor on neutrophil functions in diabetic patients with active foot infections in vitro. Twelve diabetic patients with foot infections and 12 normal volunteers were enrolled. Neutrophils from peripheral blood were incubated with granulocyte colony-stimulating factor (G-CSF, 50 ng/mL) for 20 min. Superoxide production of neutrophils was measured by the reduction of ferricytochrome C. Neutrophil phagocytosis was assayed using Staphylococcus aureus and the weighted phagocytic index was calculated. Superoxide production of neutrophils in diabetic patients with foot infections was 7.7 (unit: nmol/2 x 10(5) cells/60 min), which was significantly lower than that in controls (12.0) (p<0.05). G-CSF increased neutrophil superoxide production to 12.1 in diabetic patients with foot infections and to 19.8 in controls (p<0.05 for each). Weighted phagocytic index in diabetic patients with foot infections was 0.77, which was not significantly different from that of the controls (0.69). Weighted phagocytic index was increased significantly by G-CSF to 0.88 in diabetic patients with foot infections and to 0.79 in controls (p<0.05 for each). In conclusion, G-CSF significantly enhanced neutrophil functions in diabetic patients with foot infections in vitro.
Adult ; Aged ; Bacterial Infections/immunology* ; Diabetes Mellitus/immunology* ; Female ; Foot Diseases/immunology* ; Granulocyte Colony Stimulating Factor, Recombinant/pharmacology* ; Human ; Male ; Middle Age ; Neutrophils/immunology ; Neutrophils/drug effects* ; Receptors, IgG/analysis ; Superoxides/metabolism

No abstract available.
Adult ; Bone Marrow/pathology ; DNA Mutational Analysis ; Humans ; Leukemia, Myelomonocytic, Chronic/*diagnosis/genetics/therapy ; Leukemia, Neutrophilic, Chronic/*diagnosis/genetics/therapy ; Leukocyte Disorders/diagnosis ; Male ; Middle Aged ; Mutation ; Prognosis ; Receptors, Colony-Stimulating Factor/*genetics ; Recurrence ; *Stem Cell Transplantation ; Transplantation, Homologous

To investigate the changes of donor's peripheral blood immunocytes after mobilization with medium-dose recombinant human granulocyte-colony stimulating factor (rhG-CSF), the amounts of immunocytes in peripheral blood cells and the immunocyte components of donor peripheral blood mononuclear cells (PBMNC) in 12 healthy donors were detected by flow cytometry before and after mobilization with rhG-CSF 10 microg/(kg.day). The results showed that the median amounts of peripheral blood leukocytes before mobilization was 6.25 (4.7-7.8) x 10(9)/L, for lymphocytes it was 2.07 (1.63-3.10) x 10(9)/L, and for monocytes it was 0.163 (0.078-0.414) x 10(9)/L. In the fifth day after mobilization, the median amounts of peripheral blood leukocytes was 37.47 (24-72.57) x 10(9)/L, and for lymphocytes it was 3.22 (1.46-5.31) x 10(9)/L, and for monocytes, it was 1.2 (0.706-3.627) x 10(9)/L. The average amount of leukocytes after mobilization was 6.26 +/- 2.14 multiple of that before mobilization (P < 0.01), and the median amounts of lymphocytes after mobilization was 1.45 +/- 0.76 multiple of that before mobilization (P < 0.05), and the amount of monocytes after mobilization was 7.48 +/- 4.41 multiple of that before mobilization (P < 0.01). The median percentage of CD3(+) T lymphocytes before mobilization was 46.96% [(32.36-57.45)%], but 40.94% [(25.31-48.9)%] after mobilization. The ratio of CD4(+)/CD8(+) before mobilization was 1.27 +/- 0.46, while 1.36 +/- 0.51 after mobilization. The median percentage of CD4(+)CD8(+) T lymphocytes was 0.41% [(0.16-1.51)%], and 0.49% [(0.09-2.0)%] after mobilization. The median percentage of CD16(+)CD56(+) NK cells was 13.98% [(4.08-25.08)%] versus 16.65% [(12.06-33.05)%] after mobilization. The median percentage of CD3(+)CD16(+)CD56(+) NK-T cells was 2.75% [(0.37-6.38)%], but 3.13% [(0.46-5.95)%] after mobilization. The median percentage of CD20(+) B cells was 9.28% [(5.97-16.33)%], while 9.94% [(7.36-20.41)%] after mobilization. The median percentage of CD14(+) monocytes was 12.48% [(3.54-19.35)%] versus 29.52% [(16.51-36.76)%] after mobilization. The percentage of CD3(+) T lymphocytes, CD4(+)CD8(+) T lymphocytes, NK cells, NK-T cells and B lymphocytes in PBMNC did not change markedly before and after mobilization with middle-dose rhG-CSF. The ratio of CD4(+)/CD8(+) did not change significantly (P > 0.10). The percentages of CD14(+) monocytes in PBMNC after mobilization increased up to 2.87 +/- 1.51 higher than that before mobilization (P < 0.05). It is concluded that the changes of the CD14(+) monocytes after mobilization with rhG-CSF may be involved in graft rejection and graft versus host disease after allo-PBSCT.
Adolescent ; Adult ; Antigens, CD20 ; analysis ; Blood Donors ; CD3 Complex ; analysis ; CD4-Positive T-Lymphocytes ; cytology ; drug effects ; immunology ; CD56 Antigen ; analysis ; CD8-Positive T-Lymphocytes ; cytology ; drug effects ; immunology ; Female ; Flow Cytometry ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Lipopolysaccharide Receptors ; analysis ; Male ; Middle Aged ; Monocytes ; cytology ; drug effects ; immunology ; Peripheral Blood Stem Cell Transplantation ; Receptors, IgG ; analysis ; Recombinant Proteins

OBJECTIVETo study the changes of adhesion molecules' expressions during the recombinant human granulocyte-colony-stimulating factor (rhG-CSF) mobilization in peripheral blood stem cell transplantation (PBSCT), and to confirm the influence of rhG-CSF on hematopoietic stem cells, which are proposed to guide mobilization in PBSCT.

METHODSMice were injected subcutaneously with diluted rhG-CSF or normal saline for 7 days. The blood Sca-1(+) stem cell count and bone marrow (BM) nucleated cell count were enumerated. The expressions of CD49d and CD44 and the adhesive ability of mononuclear cells to bone marrow matrix (fibronectin) were examined by flow cytometry and (51)Cr adhesive assay, respectively.

RESULTSThe mobilizing effect of rhG-CSF on mice was the same as on humans. The number of Sca-1(+) cells in peripheral blood reached the peak on the seventh day, the BM nucleated cell count was reduced, and the expressions of CD49d and the cells' adhesive ability in BM and PB declined.

CONCLUSIONSrhG-CSF can reduce some cell adhesion molecules' expressions and the adhesive ability of hematopoietic stem cells to BM matrix, therefore mobilizing hematopoietic stem cells (HSC) from the BM to the peripheral blood.

Animals ; Bone Marrow Cells ; drug effects ; physiology ; Female ; Fibronectins ; physiology ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; Hematopoietic Stem Cell Transplantation ; Hyaluronan Receptors ; analysis ; physiology ; Integrin alpha4 ; analysis ; physiology ; Leukocytes, Mononuclear ; physiology ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins

This study was to explore the effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on neutrophil morphology, function and phenotype in patients with acute leukemia undergoing chemotherapy. Neutrophil morphology was observed under microscope with oil immersion; phagocytotic function was examined by measuring the amount of hydrogen peroxide produced by neutrophil; chemotaxis was analyzed by agarose method; oxidative burst was analyzed by flow cytometry using immunofluorescence technique; neutrophil phenotype was analyzed by flow cytometry and immunofluorescence techniques. The results showed that after rhG-CSF administration, the increased "toxic" granulation, vacuoles and Döhle bodies were observed in neutrophils of patients with acute leukemia. Compared with normal control, the functions of phagocytosis, chemotaxis, oxidative burst of neutrophil were impaired after chemotherapy, while these functions were enhanced and returned to normal level or even to be exceeded after administration of rhG-CSF. In patients with acute leukemia the neutrophil presented significantly higher expression of CD64 and CD62L than that in normal control, and a mild increase of CD64 expression and significant increase of CD62L expression were found in patients after rhG-CSF treatment. No modifications of CD16, CD32, CD14 and CD11b expression were detected in these patients before or after G-CSF administration. It is concluded that rhG-CSF administration can modify the morphology, function and phenotype of neutrophils in the patients with acute leukemia undergoing chemotherapy, and these modifications of neutrophil behavior may be supposed to be a reason for the enhancement of organism anti-infection ability.
Acute Disease ; Adult ; Aged ; Chemotaxis ; drug effects ; Female ; Flow Cytometry ; Fluorescent Antibody Technique ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Humans ; Immunophenotyping ; L-Selectin ; analysis ; Leukemia ; blood ; drug therapy ; pathology ; Male ; Middle Aged ; Neutrophils ; drug effects ; immunology ; pathology ; Phagocytosis ; drug effects ; Receptors, IgG ; analysis ; Recombinant Proteins ; Respiratory Burst ; drug effects