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At least 865 records · Page 48Linked to original sources
PubMed1989

Peripheral blood morphologic changes after high-dose antineoplastic chemotherapy and recombinant human granulocyte colony-stimulating factor administration.

The peripheral blood morphologic findings in 17 patients with cancer who had received high-dose cytotoxic chemotherapy followed by recombinant human-granulocyte colony-stimulating factor (rh-GCSF) were reviewed and compared with a control group of patients who received only high-dose chemotherapy. Both groups showed dysmyelopoiesis (abnormal granulation and nuclear lobulation) in the granulocytic series during the period of bone marrow recovery that followed the cytotoxic chemotherapy. Most of these morphologic abnormalities were more prominent in the rh-GCSF-treated group. Monocytic cells in both groups showed prominent vacuolation and immature nuclei. The percentages and absolute numbers of large granular lymphocytes were increased in the rh-GCSF group compared with the control group. No quantitative or qualitative abnormalities of eosinophilic or basophilic granulocytes were detected in either group. Both groups showed nonspecific red blood cell abnormalities, and large platelets were present in half of the control group smears. This report provides the first detailed peripheral blood morphologic description in patients treated with rh-GCSF and high-dose chemotherapy.

Adult↗
PubMed1999

Continuous-bed chromatography for the analysis and purification of recombinant human basic fibroblast growth factor.

The chromatographic properties of the commercial cation exchanger UNO-S1 (35x7 mm) was investigated using lysozyme from hen egg white as model protein and recombinant human basic fibroblast growth factor (rh-bFGF) from a high cell density cultivation of E. coli. The dynamic capacity for lysozyme (c(o) = 1 mg/ml) in 100 mM acetate buffer, pH 5 was 27 mg per ml sorbent. It was found independent of the flow-rate from 78 to 935 cm/h owing to the absence of mass transfer restrictions with this column concept. Regarding the selectivity for rh-bFGF and the capacity for lysozyme, no changes were apparent after cleaning-in-place (CIP) procedures with 0.5 M NaOH. Clogging of the column by a clarified crude cell homogenate of E. coli was not critical as precipitates were removed by reversal of the flow during CIP. Rh-bFGF elutes in three consequent peaks from the UNO-S1 column, which could be attributed to soluble rh-bFGF aggregates of different size. The dynamics of rh-bFGF aggregation and reaggregation in the crude feedstock was monitored by fast gradient elution chromatography.

Cation Exchange Resins↗
PubMed1987

Effect of recombinant human tumor necrosis factor on the colony growth of human leukemia progenitor cells and normal hematopoietic progenitor cells.

The effects of recombinant human tumor necrosis factor (rH-TNF) on the colony growth of human leukemia progenitor cells (L-CFU), granulocyte-macrophage progenitor cells (CFU-GM), and erythroid progenitor cells (BFU-E) were studied. L-CFU was assayed with leukemia cells obtained from patients with acute myelogenous leukemia. CFU-GM and BFU-E were assayed with bone marrow cells obtained from hematologically normal donors and patients with acute leukemia or non-Hodgkin's lymphoma in complete remission. A dose-dependent growth inhibition of L-CFU as well as CFU-GM and BFU-E was observed by rH-TNF at concentrations of 1 to 100 U/mL. The inhibitory effect on L-CFU was significantly greater than that on CFU-GM. No correlation was observed between the inhibitory effect on L-CFU and the number of colonies formed in the cultures without rH-TNF. Preincubation of the progenitor cells in culture medium containing 20% fetal calf serum with up to 1,000 U/mL of rH-TNF for 24 hours did not result in the inhibition of colony growth of L-CFU or CFU-GM. The inhibitory effect of rH-TNF was neutralized by an anti-rH-TNF murine monoclonal antibody.

Female↗
PubMed1990

Regulation of human eosinophil precursor production by cytokines: a comparison of recombinant human interleukin-1 (rhIL-1), rhIL-3, rhIL-5, rhIL-6, and rh granulocyte-macrophage colony-stimulating factor.

The effect of a panel of recombinant human (rh) cytokines on the generation of human eosinophil precursors was assessed using a two-step culture technique. Normal human bone marrow was preincubated with different cytokine combinations in liquid culture before assessment of the number of eosinophil progenitors, which give rise to eosinophil colony-forming units (CFU-Eo) on secondary semi-solid culture with either interleukin-5 (IL-5), IL-3, or granulocyte-macrophage colony-stimulating factor. rhIL-3 or rhGM-CSF, but not rhIL-5, increased the number of CFU-Eo. CFU-Eo production by rhIL-3 or rhGMCSF was maximal after 7 days' preincubation. Neither rhIL-1 or rhIL-6 acted on eosinophil precursors, either alone or in combination with rhIL-5, rhIL-3, or rhGM-CSF. A similar spectrum of activity of the cytokines was demonstrated whether rhIL-5, rhIL-3, or rhGM-CSF was used in the secondary cultures as the eosinophil CSF. However, rhIL-3 induced relatively more rhIL-5-responsive CFU-Eo than rhIL-3-responsive CFU-Eo, suggesting that rhIL-3 is pushing progenitors into an rhIL-5-responsive compartment.

Biological Factors↗
PubMed1989

Synergistic effects of interferon-gamma and tumor necrosis factor against proliferation of gynecologic tumor cell lines.

The cooperative effects of recombinant human tumor necrosis factor (rH-TNF) and recombinant human interferon-gamma (rH-IFN-gamma) against the proliferation of four human gynecologic tumor cell lines (HHUA, ISHIKAWA, HeLa S3, and SCH) were examined in vitro. The effect of rH-IFN-gamma on TNF receptors was also examined. ISHIKAWA and SCH cells were highly responsive to rH-TNF alone, HHUA cells exhibited a minimal degree of responsiveness to rH-TNF, and HeLa S3 cells were unresponsive to rH-TNF alone. HHUA, HeLa S3, and SCH cells were responsive dose-dependently to rH-IFN-gamma, and ISHIKAWA cells were unresponsive to rH-IFN-gamma alone. A synergistic antiproliferative effect of rH-TNF and rH-IFN-gamma appeared in HHUA, ISHIKAWA, and HeLa S3 cells. The effectiveness of rH-TNF increased in these cell lines pretreated with rH-IFN-gamma, but not vice versa. rH-IFN-gamma did not change the binding affinity of TNF receptors in the four cell lines, but increased the number of binding sites for TNF in HeLa S3 and SCH cells. These findings suggest that there is no relationship between the increase in TNF binding sites by rH-IFN-gamma and the synergistic antiproliferative effect of rH-IFN-gamma and rH-TNF. The combination therapy of rH-IFN-gamma and rH-TNF will be a new approach against cancers.

Cell Division↗
PubMed2001

Effects of recombinant human basic fibroblast growth factor on restenosis after arterial endothelial injury in rats.

AIM: To study the preventive effect of recombinant human basic fibroblast growth factor (rh-bFGF) on restenosis after percutaneous transluminal coronary angioplasty (PTCA). METHODS: Sixty male Wistar rats were randomly divided into sham operation group, intima injured group, and intima injured plus rh-bFGF treated group. Rat carotid arteries were injured using a balloon catheter except sham operation group. The rats of rh-bFGF treated group were injected im rh-bFGF 10 kU . kg-1 . d-1 after intima was injured. Ten rats in each group were killed on d 7 and d 14 after injury, respectively. [3H]Thymidine incorporation assay and pathological examination were carried out to each vessel. RESULTS: (1) Seven days after injury, the average intimal thickness in sham operation group, intima injured group, and rh-bFGF treated group was (7 +/- 1), (32 +/- 11), and (17 +/- 3) micron; average intimal area was (384 +/- 145), (1530 +/- 817), and (586 +/- 185) micron2; the numbers of smooth muscle cells in neointima per transect were 0 +/- 0, 146 +/- 18, and 105 +/- 26; the ratio of the collagen area to intimal area plus medial area were 0.29 +/- 0.09, 0.7 +/- 0.3, and 0.30 +/- 0.14; [3H]thymidine incorporation were (17 +/- 6), (62 +/- 23), (20 +/- 8) kBq/g tissue, respectively. (2) Fourteen days after injury, the average intimal thickness of sham operation group, intima injured group, and rh-bFGF treated group was (8 +/- 1), (41 +/- 9), and (20 +/- 3) micron; average intimal area was (391 +/- 134), (1761 +/- 337), and (731 +/- 124) micron2; the numbers of smooth muscle cells in neointima per transect were 0 +/- 0, 145 +/- 9, and 102 +/- 6; the ratio of collagen area to intimal area plus medial area were 0.28 +/- 0.14, 0.59 +/- 0.21, and 0.38 +/- 0.03; [3H]thymidine incorporation was (15 +/- 4), (57 +/- 11), and (22 +/- 6) kBq/g tissue, respectively. CONCLUSION: Large dosage of rh-bFGF inhibits neointimal hyperplasia and reduces restenosis after balloon injury.

Angioplasty, Balloon, Coronary↗
PubMed1999

Effect of recombinant human basic fibroblast growth factor on stomach ulcers in rats and mice.

AIM: To study the curative effects of recombinant human basic fibroblast growth factor (rh-bFGF) on gastric ulcer healing. METHODS: Pylorus ligated, water immersion stress-induced, reserpine-induced, and acetic acid-induced gastric ulcers in rats or mice were prepared. Morphometric analyses on ulcer were performed using microscope and true color image analysis system. The DNA and RNA contents were measured by diphenylamine method and orcinol method, respectively. RESULTS: In acetic acid-induced gastric ulcers in rats, rh-bFGF 2.5-40 kU.kg-1 i.g. accelerated the chronic ulcer healing with a bell-shaped dose-effect curve and the best dosage was 10 kU.kg-1. The regenerated gland epithelium width, density of capillaries in granulation tissue, and collagen content in scar tissues obviously increased in rh-bFGF-treated groups. Simultaneously, rh-bFGF promoted the differentiation and maturation of regenerated glands around ulcers. rh-bFGF 2-4 kU.kg-1 s.c. also increased the synthesis of RNA in ulcer tissues. In acute gastric ulcers, rh-bFGF i.g. was only effective on pylorus ligated ulcers, but showed no effect on total acid output and pepsin activity in gastric juice of rats. CONCLUSION: rh-bFGF promoted the gastric ulcer healing and improved the quality of gastric ulcer healing.

Acetic Acid↗
PubMed1996

Exogenous hepatocyte growth factor markedly stimulates liver regeneration following portal branch ligation in dogs.

Portal branch ligation (PBL) or embolization prior to extensive hepatectomy has been employed to increase the functional reserve of the remaining liver. This study investigated the effects of human recombinant hepatocyte growth factor (rh-HGF) on liver regeneration following PBL in dogs. Beagle dogs were subjected to PBL and were divided into two groups, a control group (n = 11) without rh-HGF and a treated group (n = 12) receiving postoperative rh-HGF at 250 ng/kg via the portal vein. Dogs were killed 72 h or 14 days following PBL. We studied the changes in serum HGF level, DNA synthesis of the liver, hepatocyte size, liver weight, and liver function tests. In the HGF group, the ratio of whole liver weight to body weight increased significantly, and both ligated and nonligated lobes showed marked increases in weight. The nonligated lobes in the HGF group showed significant increases in both DNA synthesis and hepatocyte size. Moreover, ligated lobes in the HGF group showed an increase in DNA synthesis without hypertrophy compared with the control group. Administration of rh-HGF did not significantly affect liver function tests. Ligation of the portal branch supplying the portion of liver to be resected, coupled with the administration of rh-HGF, is a useful strategy to increase hepatic reserve in advance of major hepatectomy.

Animals↗
PubMed1986

Proposal of standardized methods and reference for assaying recombinant human tumor necrosis factor.

Two assay methods for recombinant human tumor necrosis factor (rH-TNF) were developed, one a biological L-cell assay and the other an enzyme-linked immunosorbent assay. The accuracy and reproducibility of each and the correlation between the two were studied. As a result of this investigation, the two assay methods were found appropriate for standardization of rH-TNF. A freeze-dried reference was prepared, and examination of its potency and stability showed it to be suitable for use as a reference standard for rH-TNF assays.

Antibodies, Monoclonal↗
PubMed1995

[Successful treatment of cyclic neutropenia associated with hyperimmunoglobulin M syndrome using recombinant granulocyte-colony stimulating factor].

Hyper-IgM syndrome associating with severe neutropenie presenting in irregularis cycles was diagnosed in a 3-year-old male patient. His elder brother died of sepsis which appeared as a consequence of dysgammaglobulinaemia and neutropenia at the age of 9. We could not achieve a parmanent good result with the monthly immunoglobulin substitution and supportive treatment. The candida infection of the gingiva and of the oral mucous membrane expanded to the esophagus resulting in its complete occlusion and temporarily a gastrostomy was needed to feed him. We started with the recombinant human granulocyta colony stimulating factor (rh-G-CSF) treatment at the age of 6. We tried several ways of applying and finally the 10 micrograms/kg/dose given subcutan, for 5-10 days from the nadir of the neutropenic cycle seemed the most effective. The rh-G-CSF treatment resulted in an increase of ANC and the complete resolution of gingivostomatitis. The incidents of infections, the requirement of antibiotics and the duration of hospitalisation were markedly reduced. The consequent improvement in his physical condition made it possible to finally resolve the esophageal stricture surgically. We have not observed any neutropenic cycle since the end of the 14 month rh-G-CSF treatment. Though the medicine was discontinued there has been no recidiva for more than 22 months.

Candidiasis, Oral↗
PubMed1988

Synergistic cytotoxic and antitumor effects of recombinant human tumor necrosis factor and hyperthermia.

A synergistic increase in the cytotoxic effects of recombinant human tumor necrosis factor (rH-TNF) and hyperthermia was demonstrated both in vitro and in vivo. The cytotoxicity of rH-TNF against L-M cells in incubation for 12 h at 38.5 and 40 degrees C based on the concentration necessary for 50% cytotoxicity was, respectively, 125 and more than 500 times as high as in similar incubation at 37 degrees C. As observed 18 days after implantation of Meth-A fibrosarcoma cells in mice, single i.v. administration of rH-TNF at 1000 units/mouse resulted in complete cures in five mice when performed in combination with hyperthermia (40 degrees C), whereas rH-TNF alone in the same dose resulted in 27.1% inhibition of tumor growth and hyperthermia alone had no appreciable effect on tumor growth. The i.v. administration of rH-TNF three times at 100 or 300 units/mouse together with hyperthermia (40 degrees C) resulted in 41.2 and 89.0% tumor growth inhibition, respectively; similar administration without hyperthermia appeared to have little or no appreciable effect on tumor growth. The results suggest that combination therapy including rH-TNF and hyperthermia may be of value in the treatment of malignancy in human patients.

Animals↗
PubMed1994

Structure and dynamics of the human granulocyte colony-stimulating factor determined by NMR spectroscopy. Loop mobility in a four-helix-bundle protein.

Recombinant 15N- and 13C-labeled human granulocyte colony-stimulating factor (rh-metG-CSF) has been studied by 2D and 3D NMR using uniformly labeled protein, as well as residue-specific 15N-labeled samples. Assignment of 90% of the backbone resonances and 85% of side-chain resonances has enabled the determination of both the secondary and tertiary structures of the protein. The fold is similar to those of the human growth hormone and other growth factors. Four stretches of helices were identified between residues 11 and 41 (helix A), 71 and 95 (helix B), 102 and 125 (helix C), and 145 and 170 (helix D), which form a left-handed four-helix bundle with helices A and B aligned parallel to one another (up-up) and antiparallel to helices C and D (down-down). An additional short fifth helix (E) is part of the AB loop connecting helices A and B. Examination of the protein's relaxation behavior, based on the model-free approach of Lipari and Szabo, shows that the G-CSF backbone has a well-defined structure of limited conformational flexibility in helices. In contrast, the long loop connecting helices C and D exhibits substantial fast internal motion compared to the overall rotational correlation time of the whole molecule, which is on the order of 13 ns.

Chemical Phenomena↗
PubMed1993

[Wound healing of the corneal endothelium in vivo after intraocular injection of recombinant human bFGF].

In the present study, we investigated the potential role of recombinant human basic fibroblast growth factor (rh-bFGF) on the regeneration of corneal endothelium in the rabbit. For in vivo wound-healing experiments, standardized wounds were created by mechanically scraping the endothelial cells within the boundaries of a central epithelial trephine mark. A single dose of 1.5 micrograms rh-bFGF was injected into the anterior chamber of the right eye immediately after wounding, while the left eyes received the ointment vehicle only (n = 27 rabbits). Functional recovery and wound closure rates were assessed by means of ultrasonic pachymetry, cell counts after endothelial vital staining and direct computer-assisted surface analysis of Janus green-stained endothelial lesions. Measurements were carried out 0, 1, 2, 4, and 7 days after injury. For the time points day 2 and day 4, significant stimulation of endothelial regeneration in rh-bFGF treated eyes was observed with all parameters measured. These results demonstrate that rh-bFGF could prove to be useful in clinical conditions, e.g. to conserve endothelial function after intraocular surgery or as a supplement to corneal intermediate preservation media.

Animals↗
PubMed1987

The different effects of recombinant human tumor necrosis factor on rat fibrosarcoma sublines.

The antitumor effect of recombinant human tumor necrosis factor (rH-TNF) on two clones of rat fibrosarcoma with different metastatic potential to lymph nodes was examined. The colony formation of clone A, which has high metastatic potential, was completely inhibited by continuous exposure to rH-TNF at 50 U/ml. In contrast, colony formation of clone G, which has low metastatic potential, was not inhibited by high concentrations of rH-TNF (10,000 U/ml). The inhibitory effect of rH-TNF on colony formation by clone A was also observed with a 1-h exposure to rH-TNF. This effect was time and concentration dependent, as determined by the colony assay, 3H-thymidine uptake assay, and 51Cr-release assay. 3H-thymidine and 3H-uridine uptake per cell of clone A exposed to rH-TNF was not decreased. This suggests that the mechanisms of the antitumor effect of rH-TNF were not due to inhibition of DNA and RNA synthesis of tumor cells. In vivo growth and lymph node metastases of clone A inoculated i.p. to Donryu strain rats were completely suppressed by 14 consecutive i.p. injections of 10(5) or 10(6) U/kg per day of rH-TNF. On the other hand the growth of clone G was not influenced by rH-TNF administration.

Animals↗
PubMed1988

Recombinant human tumor necrosis factor-alpha. Regulation of N-formylmethionylleucylphenylalanine receptor affinity and function on human neutrophils.

Preincubation of neutrophils with recombinant human tumor necrosis factor-alpha (rH TNF-alpha) enhanced the subsequent release of superoxide anion in response to various concentrations of N-formylmethionylleucylphenylalanine (FMLP). Enhanced superoxide anion production was evident by 5 min and had reached a plateau by 15 min. Not only was the total amount of superoxide anion released greater, but the rate of release was also enhanced threefold by rH TNF-alpha. In contrast, rH TNF-alpha reduced or abolished neutrophil locomotion under agarose in response to a gradient of FMLP. Binding studies of f-Met-Leu-[3H]Phe to purified human neutrophils revealed a heterogeneous binding to unstimulated cells. The high affinity component consisted of approximately 2,000 sites per cell and had an average Kd of 2 +/- 0.7 nM (n = 4). The low affinity component consisted of approximately 40,000 sites per cell and had an average Kd of 180 +/- 50 nM (n = 4). rH TNF-alpha caused conversion to a linear Scatchard plot showing no significant change in total binding sites but a single Kd of 40 +/- 10 nM (n = 4). These data indicate that rH TNF-alpha may influence neutrophil responses to FMLP by regulating the affinity of FMLP receptors.

Cell Movement↗
PubMedDate not supplied

Augmented antitumour effects of combined treatment with hyperthermia and tumour necrosis factor on human gastric cancer xenotransplanted into nude mice.

Hyperthermia combined with recombinant human tumour necrosis factor (rH-TNF) was evaluated for antitumour efficacy in vivo. Use was made of human gastric cancer tissues xenografted into nude mice. When 100, 300, 600, and 1200 units of rH-TNF (2.4 x 10(6) units/mg protein) were given twice intraperitoneally, tumour regression did not occur in any animal. In contrast, a remarkable suppression of tumour growth was observed when 600 and 1200 units of rH-TNF was given in combination with hyperthermia at 43.5 +/- 0.1 degrees C. No effects were evident with the regimen of 100 and 300 units of rH-TNF plus hyperthermia at the same temperature, as compared with evidence obtained with hyperthermia alone. The tumoral blood flow, determined by the hydrogen diffusion method, decreased immediately after hyperthermia alone or hyperthermia plus 1200 units of rH-TNF, whereas a slight decrease was seen after rH-TNF alone. When hyperthermia plus 1200 units of rH-TNF were given, there was a remarkable delay in reversion to pretreatment values of tumoral blood flow, as compared to findings with rH-TNF only or heat only. These results are discussed in relation to the antitumour and side-effects of rH-TNF.

Animals↗
PubMed1988

Recombinant human tumor necrosis factor administered as a 24-hour intravenous infusion. A phase I and pharmacologic study.

Recombinant human tumor necrosis factor (rH-TNF) is a cytokine with direct antitumor properties. In a phase I trial we continuously infused rH-TNF for 24 hours. We gave a total of 115 courses of therapy to 50 patients. Doses ranged from 4.5 to 645 micrograms of rH-TNF/m2. Systemic toxicity, including fever, chills, fatigue, and hypotension, increased with the dose of rH-TNF administered. Doses greater than 454 micrograms/m2 frequently caused severe lethargy and fatigue, which precluded hospital discharge of the patient at the completion of therapy. The dose-limiting toxicity was hypotension, and five patients treated at the two highest dose levels required dopamine treatment. Other organ-specific toxicity was modest and spontaneously resolved after 48 hours. The 24-hour infusions of rH-TNF were associated with significant decreases in serum cholesterol and high-density lipoprotein levels. Pharmacokinetic studies using an enzyme-linked immunosorbent assay demonstrated peak plasma rH-TNF levels of 90-900 pg/mL. Despite continuous infusion of rH-TNF, no steady-state level was achieved. The recommended phase II dose for rH-TNF as a 24-hour continuous infusion is 545 micrograms/m2.

Adolescent↗
PubMed1994

A role for the carboxyl terminus of human granulocyte-macrophage colony-stimulating factor in the binding of ligand to the alpha-subunit of the high affinity receptor.

A synthetic segment (110-127) of the carboxyl terminus of recombinant human granulocyte-macrophage colony-stimulating factor (rh-GM-CSF) was used to generate a rabbit polyclonal antibody (345-6), which recognized both peptide and full-length Escherichia coli-derived rh-GM-CSF in a direct enzyme-linked immunosorbent assay. Antibody 345-6 was shown to antagonize the binding of 125I-labeled rh-GM-CSF to its receptor on the KG-1 cell line and to inhibit human GM-CSF-dependent proliferation of the AML-193 cell line. The purified IgG fraction of neutralizing antibody 345-6 was used as immunogen to obtain sheep anti-serum 1418. Antibody 1418 recognized antibody 345-6 on direct enzyme-linked immunosorbent assay but did not recognize rh-GM-CSF or the peptide 110-127 to which antibody 345-6 was raised. Antiserum 1418, as well as a purified IgG fraction of this serum, inhibited both rh-GM-CSF-stimulated cell proliferation and 125I-labeled rh-GM-CSF receptor binding but not 125I-labeled recombinant human interleukin-4 receptor binding. The anti-idiotypic antibody response derived from the anti-(110-127) antibody strongly suggests that the carboxyl-terminal region of rh-GM-CSF may be directly involved in the receptor-ligand interaction of this protein. The high affinity receptor consists of two different components (GM-R alpha beta) a cytokine-specific alpha-subunit and a beta-subunit that is shared by human GM-CSF, interleukin-3, and interleukin-5. In an effort to localize the epitope of antibody 1418 to either GMR alpha or GMR beta, several cell lines containing high, low, or both high and low affinity receptors were examined. Each was specifically and completely inhibited by antibody 1418. Interleukin-3-dependent cell proliferation of the AML-193 cell line was found to be unaffected by the antibody 1418. Thus, the carboxyl-terminal region of rh-GM-CSF is likely to be involved in the interaction of the ligand with the alpha-subunit of the high affinity receptor.

Amino Acid Sequence↗