BACKGROUNDHematopoietic stem cells (HSCs) can be used to deliver functionally active angiostatic molecules to the retinal vasculature by targeting active astrocytes and may be useful in targeting pre-angiogenic retinal lesions. We sought to determine whether HSC mobilization can ameliorate early diabetic retinopathy in mice.

METHODSMice were devided into four groups: normal mice control group, normal mice HSC-mobilized group, diabetic mice control group and diabetic mice HSC mobilized group. Murine stem cell growth factor (murine SCF) and recombined human granulocyte colony stimulating factor (rhG-csf) were administered to the mice with diabetes and without diabetes for continuous 5 days to induce autologous HSCs mobilization, and subcutaneous injection of physiological saline was used as control. Immunohistochemical double staining was conducted with anti-mouse rat CD31 monoclonal antibody and anti-BrdU rat antibody.

RESULTSMarked HSCs clearly increased after SCF plus G-csf-mobilization. Non-mobilized diabetic mice showed more HSCs than normal mice (P=0.032), and peripheral blood significantly increased in both diabetic and normal mice (P=0.000). Diabetic mice showed more CD31 positive capillary vessels (P=0.000) and accelerated endothelial cell regeneration. Only diabetic HSC-mobilized mice expressed both BrdU and CD31 antigens in the endothelial cells of new capillaries.

CONCLUSIONAuto-mobilized adult hematopoietic stem cells advance neovasculature in diabetic retinopathy of mice.

Animals ; Diabetic Retinopathy ; drug therapy ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Cell Growth Factors ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Immunohistochemistry ; Mice ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo compare the efficacy and safety of four different regimens for pediatric severe aplastic anemia (SAA) with immuno-suppressive therapy (IST) with or without combined human granulocyte colony-stimulating factor (G-CSF).

METHODThe authors retrospectively analyzed 105 children with SAA treated with IST with or without G-CSF in the hospital from February 2000 to September 2010. Regimen A, without G-CSF in the whole treatment, was used to treat Group A patients, n = 27; Regimen B, G-CSF, was initiated in Group B, n = 24, before the IST until hematologic recovery; Regimen C, G-CSF, was used together with the IST for Group C patients, n = 24, until hematologic recovery; Regimen D,G-CSF was used for Group D, n = 30, after the end of IST until hematologic recovery. The response rate, relapse rate, mortality, infection rate, infection-related death rate, risk of evolving into MDS/AML, survival rate, factors affecting the time of event-free survival and so on.

RESULT(1) The response (CR+PR) rates 4, 6, 12 and 24 months after IST of the whole series of 105 SAA children were 50.5% (7.6%+42.9%) , 60.0% (21.9%+38.1%) , 67.6% (38.1%+29.5%) and 69.5% (40.0%+29.5%) respectively. The 2-year survival rate was 90.5%; the follow-up of the patients for 13 years showed that the whole survival rate was 87.6%. (2) The differences of the response rates 4, 6, 12 and 24 months after IST of the 4 groups were not significant (P > 0.05). (3) No significant differences were found in the mortalities 4, 6, 12 and 24 months among the 4 groups (P > 0.05). (4) Of the 105 patients, 4 children had relapsed disease in the period of time from 6 to 24 months after IST. All the four patients belonged to the groups with G-CSF. (5) The use of G-CSF could not decrease the infection period before IST (day) (P = 0.273), and it had no impact on the infection rate after IST (P = 0.066). It did not reduce the rates of septicemia and infectious shock. And to the infection-related death rate no significant conclusion can be made. (6) Follow up of the patients for 13 years, showed that 2 had the evolution to MDS/AML in the 105 patients and the two children belonged to the groups with G-CSF. (7) Kaplan-meier curve analysis did not show any differences in the survival rates of the four groups. (8) Cox regression analysis showed that the use of G-CSF had no benefit to the patients' long term survival. While the age of diagnosis and the infection history before IST were significantly related to the patients' long term survival.

CONCLUSIONThe use of G-CSF did not contribute to the early response and could not reduce the infection rate, infection-related death rate and the patients' long term survival. There were no significant differences in the survival rates of the four groups. Attention should be paid to the risk of the evolution to MDS/AML.

Adolescent ; Anemia, Aplastic ; drug therapy ; immunology ; mortality ; Antilymphocyte Serum ; administration & dosage ; therapeutic use ; Child ; Child, Preschool ; Cyclosporine ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Female ; Follow-Up Studies ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; therapeutic use ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Infant ; Male ; Retrospective Studies ; Risk Factors ; Severity of Illness Index ; Survival Rate ; Treatment Outcome

Understanding of the pathophysiology of inflammatory bowel disease (IBD) is constantly evolving and, recently, a number of biologic agents have been developed. They selectively target specific molecule or pathways and correct the imbalance of the gut immune system. Among them, antibody to tumor necrosis factor (anti-TNF-alpha) is the first developed drugs, and it dramatically improved the IBD management. However, more than one-third of the patients do not respond to the drugs due to antibody formation. To increase treatment efficacy, enormous effort to develop novel anti-cytokines which can be an alternative to anti-TNF-alpha has been made. They are anti CD4+ T cell cytokine including interleukin (IL)-12/23 and IL-17 blockers, selective anti-adhesion molecule known as natalizumab, vedolizumab and alicaforsen, T-cell proliferation inhibitor, anti-inflammatory cytokine, immune stimulator, growth factor, and mitogen-activated protein kinase inhibitor. The efficacy and safety of each drugs are under investigation. Some drugs reported very promising data, however, others showed disappointing and different results. In addition, most of the trials were done in a very small number of patients, and there is no trial comparing to anti-TNF-alpha. The present paper reviews the action mechanism, short or long term efficacy and safety of variable drugs other than anti-TNF-alpha in IBD.
Antibodies, Monoclonal/therapeutic use ; Cell Migration Inhibition ; Colony-Stimulating Factors/therapeutic use ; Cytokines/antagonists & inhibitors ; Gene Therapy ; Humans ; Immunosuppressive Agents/therapeutic use ; Inflammatory Bowel Diseases/drug therapy/*therapy ; Intercellular Signaling Peptides and Proteins/therapeutic use ; Mitogen-Activated Protein Kinases/antagonists & inhibitors ; Stem Cell Transplantation ; Tumor Necrosis Factor-alpha/*antagonists & inhibitors/immunology

OBJECTIVETo investigate the optimal time for second allogeneic peripheral blood stem cell grafts (PBSC) harvest from healthy donors after in vivo recombinant human granulocyte colony-stimulating factor application (rhG-CSF).

METHODSThirty-eight healthy donors of second collection (group A) were treated with subcutaneous rhG-CSF \[5 microgxkg(-1)xd(-1)\] for five consecutive days and followed by leukapheresis on day 5 and 6. The control group (group B) was thirty-eight healthy donors who had received a first PBSC collection previously. Group A was reclassified as group C (< or = 9 months) and group D (> 9 months) according to the 75% quantile of interim time between first and second collection. The quantities of lymphocytes of CD3(+), CD3(+)CD4(+), CD3(+)CD8(+), CD14(+), CD34(+) cells and CD3(+)CD4(-)CD8(-) T cells were determined by multi-color flow cytometry.

RESULTSThe median number of CD3(+)CD8(+) (25.51 x 10(8)) and CD34(+) cells (0.51 x 10(8)) in group A were significantly lower than that (31.55 x 10(8) and 0.70 x 10(8) respectively) in group B (P < 0.05), and so did the CD3(+)CD8(+) (23.42 x 10(8)) and CD34(+) cells (0.42 x 10(8)) in group C than that in group B (P < 0.05). There was no statistical difference in median numbers of T cell subsets, monocytes, and CD34(+) cells between group B and group D (P > 0.05). The cell ratios of CD4(+)/CD8(+), CD14(+)/CD3(+) and CD3(+)CD4(-)CD8(-) T/CD3(+) in PBSC in group A, group C, and group D were similar to that in group B (P > 0.05). Sperman analysis showed a positive correlation between the total CD34(+) cells in second collection and the interval time from first to second collection (r = 0.357, P = 0.028).

CONCLUSIONNine months after the first collection maybe an optimal time for the second PBSC collection. For those who undergo second PBSC collection within 9 months, more circulation blood should be extracted to ensure enough immunological and hematopoietic compositions.

Adolescent ; Adult ; Cytapheresis ; methods ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Recombinant Proteins ; Time Factors ; Tissue Donors ; Transplantation, Homologous ; Young Adult

OBJECTIVETo investigate the cell composition in granulocyte colony-stimulating factor One (rhG-CSF) primed bone marrow grafts (G-BM) from donors with different characteristics.

METHODShundred and fifty healthy donors were injected subcutaneously rhG-CSF (5 microg x kg(-1) x d(-1)) in five consecutive days. Bone marrow and peripheral blood stem cells were harvested at day 4 and day 5. The number of CD3, CD3+ CD4+, CD3+ CD8+, CD14+ , CD34+ and CD3+ CD4- CD8- cells were determined by multicolor flow cytometry. Cell composition of G-BM from donors with different characteristics, including sex, age, weight, pregnancy were analysed.

RESULTSThe absolute number of nuclear cells (NCs), lymphocytes, CD3+, CD3+ CD4+, CD3+ CD8+, CD14+, CD34+ and CD3+ CD4- CD8- cells in G-BM were 31 050 (12 200 -58 100), 2122 (506 - 6618), 1344 (307 - 4791), 692 (145 - 3038), 616 (112 - 2575), 986 (265 -2958), 63 (11 -505) and 83 (9 -390) per microliter, respectively. The number of NCs [33 800 (18 600 - 57 100)], CD34+ cells [76 (22 -505)], and CD3+ CD4+ CD8- regulatory T cells [97 (11 - 380)] harvested from younger donors (aged < or = 38 years) were significantly higher than those from older ones (aged > 38) [28000 (12200-58100), 49 (11-220), and 65 (9 - 389) per microliter (P = 0.027, < 0.001 and = 0.001, respectively)]. Compared with that in donors with peripheral blood monocytes (PBMs) < or = 0.37 x 10(9)/L, higher number of NCs in G-BM [27 150 (13 800 - 58 100) vs 33 550 (12 200 - 57 100), P = 0.005] were collected in donors with PBMs > 0.37 x 10(9)/L. Multivariate analysis showed that donors age (< or = 38 vs > 38 years; P = 0.01) and monocyte number in peripheral blood (< or = 0.37 x 10(9)/L vs > 0.37 x 10(9)/L; P = 0. 003) were factors predicting for NC yields, and donors age ( < or = 38 vs > 38 years; P < 0.001) was factor predicting for yields of CD34+ cells (P < 0.001) and CD3+ CD4- CD8- regulatory T cells (P = 0.001) collection.

CONCLUSIONDonor age is a factor for the yields of NCs, CD34+ cells, and CD3+ CD4- CD8- regulatory T cells collection in G-BM, and donor's PBMs more than 0.37 x 10(9)/L, is another factor affecting NCs harvests.

Adolescent ; Adult ; Age Factors ; Antigens, CD34 ; Bone Marrow Cells ; immunology ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; Humans ; Male ; Middle Aged ; Recombinant Proteins ; T-Lymphocyte Subsets ; immunology ; Tissue Donors ; Young Adult

OBJECTIVE: To develop a simple method to rapidly determine the optimal time of perpheral blood stem cell harvest. METHODS: We selected 5 healthy donors mobilized with granulocyte colony-stimulating factor (G-CSF) in allogenic peripheral blood stem cell transplantation (allo-PBSCT) to continuously monitor the variety of hematopoietic stem/progenitor cells (HSC/HPC) in the peripheral blood. HPC in the peripheral blood was detected by Sysmex XE-2100 fully automated hematology analyzer with an immature myeloid information channel and the CD34+ cells and colony-forming unit-granulocyte-macrophage (CFU-GM) were determined by flow cytometry and colony culture during mobilization. RESULTS: HPC was positively correlated with the CD34+ cells or CFU-GM in the peripheral blood (r = 0.82 or r = 0.85, respectively). HPC, CD34+ cells, and CFU-GM in allo-PBSCT donors obviously increased on the 4th day after the mobilization, and their peaks were on the 5th day after the mobilization. But HPC varied more than CD34+ cells and CFU-GM did. CONCLUSION Sysmex XE-2100 can rapidly monitor the variety of HSC/HPC in allo-PBSCT donors mobilizated with G-CSF, and can provide a simple and economic method to determine the optimal time of peripheral blood stem cell harvest.
Adult ; Cell Count ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic Stem Cells ; cytology ; Humans ; Male ; Recombinant Proteins ; Time Factors

The potential therapeutic benefit of introducing IFN-gamma and GM-CSF genes in combination with the HSVtk suicide gene into subcutaneously implanted CT26 tumor cells was compared with that from each treatment alone. Cells, unmodified or retrovirally transduced with HSVtk or IFN-gamma/GM-CSF genes, were inoculated subcutaneously into syngeneic BALB/c mice in various combinations. HSVtk gene, with intraperitoneal ganciclovir treatment, reduced tumor volume by 81% at locally inoculated tumor sites (p<0.01) and by 25% at distantly inoculated tumor sites (p=0.052). IFN-gamma/GM-CSF genes showed a 56% tumor volume reduction at local tumor sites (p<0.01) and 15% volume reduction at remote tumor sites, although this was not statistically significant. The combination of HSVtk (with GCV) and IFN-gamma/GM-CSF genes showed an 81% volume reduction at local tumor sites (p<0.01) and a 43% volume reduction at remote tumor sites (p<0.01). Thus, the combination of HSVtk and IFN-gamma/GM-CSF gene therapy produced greater therapeutic efficacy than either treatment alone.
Animals ; Cell Line ; Cell Line, Tumor ; Gene Therapy/*methods ; Genes, Transgenic, Suicide ; Granulocyte Macrophage Colony-Stimulating Factors, Recombinant/biosynthesis/*genetics/therapeutic use ; H-2 Antigens/metabolism ; Interferon-gamma, Recombinant/biosynthesis/*genetics/therapeutic use ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasms, Experimental/*therapy ; Research Support, Non-U.S. Gov't ; Simplexvirus/enzymology/genetics ; Thymidine Kinase/*genetics/therapeutic use ; Transduction, Genetic

Although enumeration of CD34+ cells in the peripheral blood (PB) on the day of apheresis predicts the quantity of those cells collected, the flow cytometric techniques used are complex and expensive, and several hours are required to obtain the result in the clinical practice setting. The Sysmex SE-9000 automated haematology analyzer provides an estimate of immature cells, called hematopoietic progenitor cells (HPC). The aim of this study was to evaluate the clinical usefulness of HPC in predicting the optimal timing of peripheral blood progenitor cells (PBPC) harvest. Studies were performed on 628 aphereses from 160 patients with hematologic or solid malignancies. Spearman's rank statistics was used to assess correlation between HPC, WBC, mononuclear cells (MNC), and CD34+ cells. A receiver operating characteristic (ROC) curve was drawn for cutoff value of HPC, and predictive values of the chosen cutoff value of HPC for different target CD34+ cell collections were calculated. The PB HPC had a stronger correlation (rho=0.592, p<0.001) with collected CD34+ cells than did PB WBC and PB MNC. The ROC curve showed that the best cutoff value of HPC was 50x10(6)/L for the target CD34+ cells >or=1 x10(6)/kg with sensitivity of 75%. Positive and negative predictive values of HPC >or=50x10(6)/L for CD34+ cells >or=1x10(6)/kg were 59.7% and 81.1%, respectively. In the clinical practice setting, applying variable cutoff values of HPC would be a useful tool to predict the optimal timing of PBPC collection.
Adolescent ; Adult ; Antigens, CD34/analysis ; Antineoplastic Combined Chemotherapy Protocols/pharmacology ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use ; Blood Cell Count*/instrumentation ; Combined Modality Therapy ; Female ; Granulocyte Colony-Stimulating Factor/pharmacology ; Hematopoietic Stem Cell Mobilization* ; Hematopoietic Stem Cells* ; Human ; Leukapheresis* ; Male ; Middle Aged ; Neoplasms/blood ; Neoplasms/drug therapy ; Neoplasms/therapy ; Peripheral Blood Stem Cell Transplantation* ; Predictive Value of Tests ; Sensitivity and Specificity ; Time Factors

Osteoclasts are bone-specific multinucleated cells generated by the differentiation of monocyte/macrophage lineage precursors. Regulation of osteoclast differentiation is considered an effective therapeutic approach to the treatment of bone-lytic diseases. Periodontitis is an inflammatory disease characterized by extensive bone resorption. In this study, we investigated the effects of sodium fluoride (NaF) on osteoclastogenesis induced by Porphyromonas gingivalis, an important colonizer of the oral cavity that has been implicated in periodontitis. NaF strongly inhibited the P. gingivalis-induced alveolar bone loss. That effect was accompanied by decreased levels of cathepsin K, interleukin (IL)-1β, matrix metalloproteinase 9 (MMP9), and tartrate-resistant acid phosphatase, which were up-regulated during P. gingivalis-induced osteoclastogenesis. Consistent with the in vivo anti-osteoclastogenic effect, NaF inhibited osteoclast formation caused by the differentiation factor RANKL (receptor activator of nuclear factor κB ligand) and macrophage colony-stimulating factor (M-CSF). The RANKL-stimulated induction of the transcription factor nuclear factor of activated T cells (NFAT) c1 was also abrogated by NaF. Taken together, our data demonstrate that NaF inhibits RANKL-induced osteoclastogenesis by reducing the induction of NFATc1, ultimately leading to the suppressed expression of cathepsin K and MMP9. The in vivo effect of NaF on the inhibition of P. gingivalis-induced osteoclastogenesis strengthens the potential usefulness of NaF for treating periodontal diseases.
Acid Phosphatase ; drug effects ; Alveolar Bone Loss ; microbiology ; prevention & control ; Animals ; Anti-Bacterial Agents ; therapeutic use ; Anti-Inflammatory Agents ; therapeutic use ; Bacteroidaceae Infections ; microbiology ; prevention & control ; Bone Density Conservation Agents ; therapeutic use ; Cathepsin K ; drug effects ; Interleukin-1beta ; drug effects ; Interleukin-6 ; analysis ; Interleukin-8 ; drug effects ; Isoenzymes ; drug effects ; Macrophage Colony-Stimulating Factor ; drug effects ; Male ; Matrix Metalloproteinase 9 ; drug effects ; Osteoclasts ; drug effects ; Periodontitis ; microbiology ; prevention & control ; Porphyromonas gingivalis ; drug effects ; RANK Ligand ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sodium Fluoride ; therapeutic use ; Tartrate-Resistant Acid Phosphatase ; Transcription Factors ; drug effects ; X-Ray Microtomography ; methods