Granulocyte-Macrophage Colony-Stimulating Factor ; therapeutic use ; Humans ; Recombinant Fusion Proteins ; therapeutic use

This study was purposed to evaluate the long-term outcome and the safety of autologous peripheral blood mononuclear cells (PBMNC) treated by interleukin 2 (IL-2) and granulocyte-macrophage colony stimulating factor (GM-CSF) in the therapy of patients with aplastic anemia (AA). The therapy of 49 patients admitted BG in hospital from April 2001 to December 2007 were analyzed retrospectively. PBMNC were isolated and cultured for 48 hours in presence of IL-2 and GM-CSF. Cells were collected, and 6 × 10(6) - 1 × 10(8) PBMNC were intravenously injected weekly for 4 - 22 months. Hematopoietic recovery was evaluated by examinations of peripheral blood, bone marrow aspirates and bone marrow biopsy. Flow cytometry was used to assess the peripheral T cell subsets before and after treatment. Polymerase chain reaction was performed to observe the clonal diversity of T cell receptor variable β-chain (TCR-Vβ) recombination. The results showed that 37 cases were cured and none of them relapsed during the follow-up, 5 cases were in partial remission, 3 cases got improvement, and 4 cases showed no response. The total efficiency reached up to 91.8%. The ratios of CD4(+)/CD8(+) subsets were abnormal in 39 patients prior to the treatment, and 31 cases restored to the normal range after cell transfusions. Analysis on the clonal diversity of TCR-Vβ recombination in 11 patients showed the transition from monoclonal or biclonal spectratype to polyclonal one. No long-term side effects were documented. It is concluded that the treatment with PBMNC treated by IL-2 and GM-CSF is generally safe and effective. The underlying mechanisms may be in relation to the restoration of cell immunity.
Adolescent ; Adult ; Anemia, Aplastic ; therapy ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Granulocyte-Macrophage Colony-Stimulating Factor ; therapeutic use ; Humans ; Interleukin-2 ; therapeutic use ; Male ; Middle Aged ; Monocytes ; transplantation ; Peripheral Blood Stem Cell Transplantation ; methods ; Transplantation, Autologous ; Young Adult

OBJECTIVETo construct a recombinant plasmid pIRES-GM-CSF-IL-21, and to investigate its antitumor effect on tumors in the mice.

METHODSFifty Bal b/c mice were included in this study. Cultured hepatoma H22 cells were inoculated in the left lobe of the liver to develop orthotopically transplanted liver tumor models. The mice with orthotopically transplanted liver tumor were randomly divided into 5 groups (n = 10): (1) Each mouse received injection of recombinant plasmid pIRES-GM-CSF-IL-21; (2) Received injection of plasmid pIRES-GM-CSF; (3) pIRES-IL-21; (4) Received injection of ampty plasmid pIRES (H22/neo group); (5) Received injection of PBS (H22 group) via the tail vein, respectively. Therefore, the anti-tumor effect was induced by both GM-CSF and IL-21, or by either of them alone. The serum levels of IFN-γ and IL-2 were detected by ELISA, and the cytotoxicity of spleen NK and CTL cells were tested by MTT colorimetry.

RESULTSComparing with the H22 and H22/Neo groups, the tumor weight in the mice of H22/GM-CSF group was (0.603 ± 0.223) g, H22/IL21-treated group (0.583 ± 0.290) g and H22/GM-CSF-IL21-treated group (0.303 ± 0.323) g, significantly lower than that in the H22 group [(1.591 ± 0.280) g] and H22/Neo group [(1.489 ± 0.155) g]. Among them the tumor growth was most significantly inhibited in the H22/GM-CSF-IL-21 group (0.303 ± 0.323) g, compared with that of H22 and H22/neo groups (P < 0.01). But there was no significant difference between the tumor weights of the H22/GM-CSF group and H22/IL-21 group, and between the tumor weights of the H22 and H22/Neo groups (P > 0.05). The levels of IFN-γ and IL-2 in peripheral blood of mouse models treated with H22/GM-CSF-IL-21 were significantly increased than that in the H22/GM-CSF group and H22/IL-21 group (all P < 0.01), but significantly decreased in the H22group and H22/Neo group (P < 0.01). The anti-tumor activity of splenic NK cells and CTLs in the H22/GM-CSF-IL21 group was significantly enhanced (P < 0.01), compared with the significantly decreased in the H22 and H22/Neo groups.

CONCLUSIONSOur results demonstrate apparent antitumor effect of the plasmid pIRES-GM-CSF-IL-21 on the orthotopically transplanted liver tumor in mice. The combination of both pIRES-GM-CSF and IL-21 is more effective than that of pIRES/IL21 or pIRES/GM-CSF treatment alone. In addition, the plasmid pIRES-GM-CSF-IL-21 can also promote the secretion of IFN-γ and IL-2 in vivo, and enhance the cytotoxic activity of splenic NK and CTLs against the transplanted liver tumor.

Animals ; Carcinoma, Hepatocellular ; blood ; pathology ; therapy ; Cell Line, Tumor ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Immunotherapy ; Interferon-gamma ; blood ; Interleukin-2 ; blood ; Interleukins ; genetics ; Killer Cells, Natural ; immunology ; Liver Neoplasms ; blood ; pathology ; therapy ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Plasmids ; therapeutic use ; Random Allocation ; Recombinant Proteins ; therapeutic use ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Burden

UNLABELLEDOBJECTIVE To explore the effect of combined use of rehmannia (RM) and rhodiola (RD) on peripheral leukopenia and bone marrow hematopoietic function suppression induced by cyclophosphamide (CTX) in mice.

METHODSICR mice were established into bone marrow inhibition models by intraperitoneal injection of CTX, and were administered with RM, RD or its extract (RDE), singly or in mixture, via gastrogavage for 10 days. The changes of peripheral hemogram, bone marrow nucleated cell proliferation, CFU-GM colony formation, GM-CSF and erythropoietin (EPO) secretion were observed.

RESULTSCompared with the un-treated model mice, the peripheral white blood cell count was significantly higher in model mice treated with RDE and RM mixture; the bone marrow nucleated cells count, CFU-GM formation, and GM-CSF production were significant higher in model mice treated with RD and RM mixture, showing statistical significance (P < 0.01); while EPO production in the RD and RM mixture treated group was slightly elevated, but the difference showed no statistical significance.

CONCLUSIONRD and RM mixture could regulate hematopoietic system by promoting the production of bone marrow cells and colonies, as well as enhancing the synthesis of related cytokines, such as GM-CSF, so as to increase the amount of peripheral white blood cells and restore the hematopoietic function of organism.

Animals ; Bone Marrow ; drug effects ; Cyclophosphamide ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Erythropoietin ; secretion ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; secretion ; Hematopoiesis ; drug effects ; Leukopenia ; chemically induced ; drug therapy ; Male ; Mice ; Mice, Inbred ICR ; Phytotherapy ; Rehmannia ; chemistry ; Rhodiola ; chemistry

OBJECTIVETo evaluate the efficacy and safety of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) hydrogel in wound healing in patients with deep partial thickness burn.

METHODSThe study was a multicenter, randomized, double-blind, placebo-controlled parallel clinical trial. Three hundred and twenty-one patients (302 cases finally fulfilled the protocol) with deep partial thickness burn were divided into A group (n = 200, with treatment of rhGM-CSF hydrogel, 100 microg/10 g/100 cm2/d), C group (n = 102,with treatment of placebo). Side-effect, systemic condition, wound healing time, wound healing rate, and total effective rate at different time points were observed.

RESULTSThere were no obvious differences in vital signs, wound secretion, wound edge reaction, blood and urine routine, liver and kidney function between two groups (P > 0.05). No side-effect was observed. The median wound healing time was 17 days in A group, which was obviously shorter than that in C group (20 days, P < 0.01). The mean wound healing rate in A group was 24.5%, 70.5%, 95.3%, 99.6% respectively on 8th, 14th, 20th, 28th day after treatment, which were obviously higher than that in C group (15.1%, 51.4%, 84.6%, 97.1%, respectively, P < 0.01). The total effective rates in A group on 8th, 14th, 20th day after treatment were also higher than that in C group (P < 0.01).

CONCLUSIONrhGM-CSF hydrogel can significantly accelerate wound healing in patients with deep partial thickness burn with certain safety.

Burns ; drug therapy ; Double-Blind Method ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; adverse effects ; therapeutic use ; Humans ; Hydrogels ; therapeutic use ; Male ; Placebos ; Recombinant Proteins ; Wound Healing

OBJECTIVETo investigate the anti-tumor effects and mechanism of tumor vaccines and whether chemotherapeutic agents administered prior to immunotherapy could augment the efficacy of the vaccines.

METHODSC57/BL mice inoculated with Lewis lung cancer cells were used as tumor models. Granulocyte-macrophage colony-stimulating factor (GM-CSF) gene modified LA795 and Lewis lung cancer cell lines were administered as allogeneic and autologous tumor vaccines, respectively. After Lewis cells (1 x 10(7)) inoculation, the mice received irradiated GM-CSF secreting cancer vaccine solely or in combination with carboplatin. The survival of the mice was observed. The cytotoxicity of spleen cells or purified CD8(+) cells was analyzed by lactate dehydrogenase (LDH) assay. Serum level of IL-4 and IFN-gamma was detected using ELISA method.

RESULTSThe cytotoxicity of the spleen cells or purified CD8(+) T cells against Lewis cells in the mice immunized with cancer cell vaccine was significantly increased, relative to that of the control, untreated group (P < 0.05). Serum level of Th1-type cytokine IFN-gamma was increased after vaccination, whereas Th2-type cytokine IL-4 showed no significant change. The GM-CSF secreting cancer cell vaccine had no significant influence on the survival of the mice with established heavy tumor burden. The combination of chemotherapy and cancer vaccine could statistically prolong the survival time; whereas any method itself had no significant effect.

CONCLUSIONThe GM-CSF secreting cancer cell vaccine can induce immune responses. The chemotherapeutic agents may be beneficial to enhance the anti-tumor activity of cancer vaccine.

Animals ; Antineoplastic Agents ; therapeutic use ; Cancer Vaccines ; therapeutic use ; Carboplatin ; therapeutic use ; Carcinoma, Lewis Lung ; blood ; metabolism ; pathology ; therapy ; Cell Line, Tumor ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; metabolism ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Lung Neoplasms ; metabolism ; pathology ; Mice ; Mice, Inbred C57BL ; Transfection

OBJECTIVETo observe the therapeutic efficacy of 131I-labeled granulocyte macrophage colony-stimulating factor (GM-SCF) in SCID mouse-acute myeloid leukemia model, and the relationship between dose and effect.

METHODSSCID-mouse acute myeloid leukemia model was established by injecting HL-60 cells through tail vein. GM-CSF was labeled with 131I by the chloramines-T method. SCID mice were randomly divided into 6 groups. Groups I, II and III treatment groups were given 9.25 x 10(5), 22.20 x 10(5) and 37.00 x 10(5) Bq of 131I-GM-CSF, respectively. Group IV was given 131I. Group V was given blending of 131I and GM-CSF. Group VI was control. Changes of HL-60 cells in blood and marrow, as well as white blood cells, red blood cells and platelets in blood were detected. Survival time of the SCID mice was calculated.

RESULTIt was observed that WBC, HL-60 cells in blood and marrow were less in treatment groups than that in control groups, especially in groups II, III. After 2 weeks of treatment, BPC of II, III groups increased remarkably (P < 0.01). Survival time of the SCID mice was prolonged in treatment groups (P < 0.01), especially in group III, the longest survival time of 60 days.

CONCLUSION131I-GM-CSF could increase leukemic SCID mice survival rate. The therapeutic efficacy of low dose and mediate dose of 131I-GM-CSF is dose-dependent. 131I-GM-CSF is an effective radiation immunity therapy for leukemic mice.

Animals ; Antineoplastic Agents ; therapeutic use ; Dose-Response Relationship, Drug ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; therapeutic use ; HL-60 Cells ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; Mice ; Mice, SCID ; Xenograft Model Antitumor Assays

OBJECTIVETo evaluate the effects of autologous tumor vaccines in preventing recurrences of hepatocellular carcinoma (HCC).

METHODSFrom March 1999 to June 2003, 80 patients with HCC undergoing resections were randomly assigned into a tumor vaccine group (n=40) and a control group (n=40). Tumor vaccines, consisting of formalin-fixed HCC tissue fragments, biodegradable sustained-releasers of granulocyte-macrophage-colony stimulating factor, interleukin-2, and an adjuvant, were developed. Every vaccine group patient received 3 vaccinations at a 2-week interval and the control group just received the adjuvant. Delayed-type-hypersensitivity (DTH) test and recurrent rates were analyzed.

RESULTSEight patients of the vaccine group and five patients of the control group were lost in the follow-up. Thirty-two patients completed the tumor vaccine procedure and no essential adverse effects occurred. 23/32 patients developed DTH responses against the fragments of HCC. The follow-up averaged 34.3 months (from 15 to 55 months). 1-, 2-, 3-year recurrence rates of the vaccine group were 12.6%, 35.9% and 54.0%, respectively; 1-, 2-, 3-year recurrence rates of the control group were 31.6%, 61.3% and 72.1%, respectively. The recurrent rate was significantly better in the tumor vaccine group than in the control group (P = 0.037).

CONCLUSIONSAutologous tumor vaccine is a promising adjunctive modality to prevent recurrence of human HCC.

Adult ; Aged ; Cancer Vaccines ; therapeutic use ; Carcinoma, Hepatocellular ; surgery ; therapy ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; therapeutic use ; Humans ; Interleukin-2 ; therapeutic use ; Liver Neoplasms ; surgery ; therapy ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Postoperative Period ; Vaccination

OBJECTIVETo compare the effects of puerarin and granulocyte colony-stimulating factor (G-CSF) in treating acute myocardial infarction (AMI) and on the size of infarcted area and cytokines.

METHODSSeventy-nine patients with anterior AMI were randomly divided into three groups, they were treated with conventional Western medical treatment, but to the puerarin group (PG) and the G-CSF group (GCG) puerarin and G-CSF was given additionally, respectively. The infarcted size, plasma G-CSF, matrix metalloproteinase-9 (MMP-9), serum interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were detected before and after treatment.

RESULTSThe infarcted size was positively correlated to the levels of G-CSF, MMP-9, IL-6 and TNF-alpha before treatment ( r = 0.45, 0.42, 0.44 and 0.42, P<0.01 ). The infarcted size in the PG and the GCG decreased on the 28th day (P<0.01), the level of G-CSF, MMP-9, IL-6 and TNF-alpha in the PG on the 7th day all decreased (P<0.05), but these indexes in the GCG increased (P<0.05), while those in the control group were unchanged (P>0.05).

CONCLUSIONPuerarin and G-CSF are effective in decreasing infarcted size, but their effects on cytokines are different entirely.

Aged ; Antigens, CD34 ; metabolism ; Drug Therapy, Combination ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; therapeutic use ; Humans ; Interleukin-6 ; blood ; Isoflavones ; therapeutic use ; Male ; Matrix Metalloproteinase 9 ; blood ; Middle Aged ; Myocardial Infarction ; drug therapy ; Phytotherapy ; Recombinant Proteins ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate clinical effects of dendritic cells (DCs) pulsed with autologous hepatoma cell lysates on postoperative recurrence and metastasis of hepatocellular carcinoma (HCC).

METHODSDCs isolated from peripheral blood mononuclear cells of HCC patients were cultured and propagated in vitro using rhGM-CSF and rhIL-4, and then were pulsed with autologous hepatoma cell lysates. Thirty postoperative patients with HCC were randomly divided into two groups. Fifteen cases were treated with DC vaccine; fifteen cases received chemotherapy only as a control group. Immune function, clinical effects, hepatic tumor recurrence rate and the survival rate of patients of the two groups were observed and compared.

RESULTSThe levels of CD3+, CD4+/CD8+ and NK cells in the DC vaccine group were significantly increased after vaccination, while those of the control group had no significant changes. The concentration of IL-10 in the DC vaccine group was significantly decreased after the vaccination (P < 0.05). The hepatic tumor recurrence rate at 18 months in the DC vaccine group was 13.33%, compared with 53.33% in the control group (P < 0.05). The survival rate in the former was 93.33%, compared with that of 60% in the later (P < 0.05).

CONCLUSIONHepatoma cell lysates pulsed DC vaccine may improve the immune function of the postoperative HCC patients and play an important role in prevention of postoperative recurrence and metastasis of HCC, which would provide an innovative approach for the immunotherapy of HCC.

Cancer Vaccines ; therapeutic use ; Carcinoma, Hepatocellular ; pathology ; therapy ; Dendritic Cells ; immunology ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; therapeutic use ; Humans ; Immunotherapy ; Interleukin-4 ; therapeutic use ; Liver Neoplasms ; pathology ; therapy ; Male ; Middle Aged ; Neoplasm Metastasis ; prevention & control ; Neoplasm Recurrence, Local ; prevention & control ; Postoperative Period ; Recombinant Proteins ; therapeutic use