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1.
Summary Antibody-dependent cell-mediated cytotoxicity (ADCC) mediated by a murine monoclonal antibody against human colerectal carcinoma, antibody 19–9, with human effector cells was tested in 33 patients with various carcinomas, 16 patients with benign lesions, and 13 normal controls, using a 12-h 51Cr release assay using human colorectal cancer cells as targets. Peripheral blood mononuclear cells (PBM) from these groups of patients and normal controls achieved moderate levels of target cell lysis in the presence of the monoclonal antibody at the high effector to target cell ratio of 200:1. The ADCC activity of PBM in cancer patients was significantly higher than that in either normal persons or patients with benign lesions. Since the ADCC was shown to be mainly mediated by adherent monocytes in the PBM, ADCC activity of monocytes from cancer patients was compared to those from control groups at an effector to target cell ratio of 30:1. The results also showed that the lytic capacity of monocytes was significantly higher in cancer patients than that in the control populations.  相似文献   

2.
The embryonic stem cell line, S25, is a genetically modified line that allows lineage selection of neural cells (M. Li, L. Lovell-Badge, A. Smith (1998) Current Biology 8: 971–974). Here, the growth parameters of this cell line were analysed. Serial passaging in adherent conditions enabled these cells to grow rapidly (average specific growth rates of 0.035 h–1) and generate high viable cell densities (above 90%). The aggregation of the S25 cells into embryoid bodies (EBs) was also studied, indicating limited cell growth (maximum cell densities of 2.7×105 cells ml–1) and a high variability of aggregate size (70–400 m after 8 d). Enzymatic dissociation of EBs with 1% (v/v) trypsin gave highest cell viability (91%) and density (1.4×104 cells ml–1) and the cells thus obtained are able to differentiate into neurons.  相似文献   

3.
Summary The administration of a thymic factor, thymostimulin (TP-1), to mice resulted in considerable augmentation of natural killer (NK) cell activity as measured in a short-term assay against 51Cr-labeled YAC-1 target cells. Conditions suitable for detection of the thymostimulin-induced boosting of NK included multiple daily exposures to TP-1 (50 g/kg), and peak levels of reactivity were observed at 2–4 days after discontinuation of treatment. A strict age-dependency of the effect was also observed, with optimal augmentation of NK-cell activity when TP-1 was administered to mice at 4–6 weeks of age. The effect was not limited to TP-1 treatment but was also observed on administration of another thymic factor (thymosin 1). The activated cells responsible for the increased natural cell-mediated cytotoxicity appeared to be typical murine NK cells, judging by both functional and antigenic criteria.  相似文献   

4.
Superantigen-based tumor therapy: in vivo activation of cytotoxic T cells   总被引:23,自引:0,他引:23  
We have recently demonstrated that the superantigen staphylococcal enterotoxin A (SEA) targets in vitro activated cytotoxic T lymphocytes against tumor cells expressing major histocompatibility complex (MHC) class II antigens. In this report we analyze the use of SEA as an immunoactivator in vivo. Treatment of mice with SEA activated a fraction of CD3+ T cells apparently as a function of their T cell receptor V expression. SEA induced interleukin-2 receptor expression and proliferation in both CD4+ and CD8+ T cells. This proliferative response was dose-dependent (0.1 – 100 µg/mouse), peaked during day 1 after treatment and declined to background levels within 4 days. The cytotoxic response, measured as cytotoxicity to SEA-coated MHC class II+ target cells (staphylococcal-enterotoxin-dependent cell-mediated cytotoxicity, SDCC), was maximal at a dosage of 1 µg SEA/mouse. The SDCC was confined to the CD8+ T cell compartment, peaked 2 days after treatment and declined to background levels within 4 days. A second injection of SEA on day 5 after the first SEA treatment resulted in SDCC function with kinetics and magnitude identical to that seen after one injection. These results pave the way for the use of SEA in the treatment of MHC class II+ tumors.  相似文献   

5.
Synopsis Embryos in specific stage of the estuarine teleost, Fundulus heteroclitus, were exposed to mercuric chloride (MC) and methylmercuric chloride (MMC) under several distinct treatment conditions. Four-eight cell stage eggs (0-day old) were exposed for 4 days (continuous), 2 days and one day to each mercury compound. One-day old (mid-blastula), 2-day old (mid-neurula) and 5-day old (beating heart) embryos were exposed 4 days to MC and MMC. Mortality for the four days immediately following the initiation of exposure was the embryonic response measured. Under most exposure conditions to the 4–8 cell eggs, progressive and significant reductions in survival were observed at all concentrations above 40 and 30 gHg++l–1 as MC and MMC, respectively. Reducing the duration of exposure to 1 day most significantly increased the survival potential of the 4–8 cell eggs. For all exposure treatments to the 4–8 cell eggs, significant differences in survival, between eggs exposed to MC and MMC, were determined at 40, 60 and 80 gHg++l–1, indicating the presence of compound-dependent response differences. In all cases demonstrating response differences between MC and MMC exposed embryos, survival was significantly lower following exposure to MMC. Survival of embryos was progressively increased when the initiation of continuous exposure (4 days) was delayed 1, 2 and 5 days after fertilization. As a result, compound-dependent response differences were progressively shifted to higher He++ concentrations. For both MC and MMC, survival of 1-day old embryos exposed for 4 days was greater than that of 0-day old eggs exposed for 1 day. Of the embryonic stages examined, it appears that the earlier cleavage stages are the most sensitive to mercury intoxication.  相似文献   

6.
Summary Human peripheral blood mononuclear cells (lymphocytes and monocytes) were preincubated for 0–24 h with human recombinant granulocyte-monocyte-colony-stimulating factor (GM-CSF) and used as effector cells in an 18 h antibody-dependent cellular cytotoxicity (ADCC) assay with SW948 (a human colorectal carcinoma cell line) as target cells and mAb 17-1A. A significant increase in the lytic capability was noted after 0.5–2 h of preactivation while longer preincubation times did not significantly increase the lytic potential. GM-CSF at 0.01 g/ml induced the best tumor cell lysis while higher concentrations were inhibitory. GM-CSF pretreatment induced a statistically significant increase in the lytic capacity of both monocytes and lymphocytes in ADCC as well as in the spontaneous cytotoxicity.  相似文献   

7.
Summary Chaetomium cellulolyticum (ATCC 32319) was cultivated on glucose, Avicel and/or Sigmacell in a 20-1 stirred tank batch reactor. The substrate (cellulose) concentration, the cell mass concentration (through protein and/or nitrogen content), reducing sugar concentration, the enzyme activity, the alkali consumption rate, the dissolved O2 and CO2 concentrations in the outlet gas were measured. The specific growth rate, the substrate yield coefficient, cell productivity, the oxygen consumption rate, the CO2 production rate and the volumetric mass transfer coefficient were determined. At the beginning of the growth phase the oxygen utilization rate exhibits a sharp maximum. This maximum could be used to start process control. Because of the long lag phase periodic batch operation is recommended.Symbols CP cell protein concentration (g l–1) - FPA FP enzyme activity (IU l–1) - GP dissolved protein concentration (g l–1) - IU international unit of enzyme activity - kLa volumetric mass tranfer coefficient (h–1) - LG alkali (1 n NaOH) consumption (ml) - LGX specific alkali consumption rate per cell mass (ml g–1 h–1) - P cell mass productivity (g l–1 h–1) - specific oxygen consumption rate per cell mass (g g–1 h–1) - Q aeration rate (volumetric gas flow rate per volume of medium, vvm) (min–1) - N impeller speed (revolution per minute, rpm) (min–1) - S substrate concentration (g l–1) - S0 S at tF=0 (g l–1) - S0 S in feed (g l–1) - SR acid consumption (ml) - TDW total dry weight (g l–1) - T temperature (° C) - tF cultivation time (h) - U substrate conversion - X cell mass concentration (g l–1) - YX/S vield coefficient - specific growth rate (h–1) - m maximum specific growth rate (h–1)  相似文献   

8.
Summary Plasmin, a serine protease, was recently found to be involved in corneal ulcerative processes in humans and rabbits. In our experiments, plasmin activity was found in the tear fluid after mechanical and chemical damage of the rabbit cornea, such as de-epithelization and burning with alkali. The plasmin concentrations in the tear fluid were dependent on the severity of injury. The highest plasmin activity (2.0–3.0 g ml–1) occurred after severe alkali damage to large areas of the cornea, and the lowest activity (0.4–1.0 g ml–1) after mechanical injury (de-epithelization).Plasmin concentrations up to 1.0 ml–1 were associated with increased activities of lysosomal hydrolases in epithelial cells and keratocytes beneath the epithelium. Plasmin activities increased as the inflammatory reaction developed. When plasmin activity in the tear fluid was higher than 1.0 g ml–1, inflammatory cells were found in the corneal stroma. Levels of 1.5–2.0 g ml–1 were connected with higher numbers of inflammatory cells (particularly polymorphonuclear leukocytes) with increased activities of lysosomal hydrolases. Very high plasmin activities (2.5–3.0 g ml–1) accompanied corneal ulcerative processes.The local application of aprotinin (Trasylol, Bayer), an inhibitor of plasmin, and also of some other proteases, was found to be necessary for the healing of severe corneal injuries in which highly elevated plasmin activity in the tear fluid and inflammatory cellulization of the cornea occurred (severe damage). It was beneficial in cases in which medium plasmin activity occurred in the tear fluid and inflammatory changes in the cornea were not too extensive. If used very early after injury, aprotinin prevents the appearance of high plasmin activity in the tear fluid, reduces the invasion of inflammatory cells into the corneal stroma, and accelerates the healing. Even the corneal transparency is restored in many cases.  相似文献   

9.
Recently, cadmium has been described to disturb ovarian function in rats. In this paper the direct influence of cadmium on steroid production of ovarian cellsin vitro has been studied. Granulosa and luteal cells were obtained from proestrous and pregnant rats, and incubated with 0, 5, 10, 20 or 40 g ml–1 CdCl2 in the presence or absence of 0.1–1000 ng ml–1 follicle stimulating hormone (FSH) or luteinizing hormone (LH) for 24 or 48 h. Production of progesterone (P) and 17-estradiol (E2) by granulosa and that of P by luteal cells were measured by radioimmunoassay. In FSH-stimulated granulosa cell cultures, 5 and 40 g ml–1 CdCl2 suppressed P accumulation to 65 and 10%, respectively; accumulation of E2 (at 5 g ml–1 CdCl2) decreased to 44%. P production of LH-supported luteal cells dropped to 86 and 66%, respectively, when 5 and 40 g ml–1 CdCl2 was added to the medium. No alteration in basal P accumulation occurred in granulosa and luteal cell cultures following incubations with 20 and 40 g ml–1 CdCl2, whereas basal E2 production of granulosa cells was markedly diminished. It is concluded that CdCl2 suppressing steroid synthesisin vitro exerts a direct influence on granulosa and luteal cell function.  相似文献   

10.
Epidemiological, experimental and clinical data indicate that cadmium and lead are osteotoxins in man and other species. The relative sensitivities of a clonal human osteosarcoma cell line (HOS TE 85) and a clonal rat osteosarcoma cell line (ROS 17.28) to the cytotoxic effects of cadmium and lead were tested in serum-free media without added growth factors. The rat osteosarcoma cells were more sensitive to cadmium with cytotoxicity and inhibition of proliferation at 0.25 versus 0.75 and 1.0 mol l cadmium, respectively, for human osteosarcoma cell lines. The lower sensitivity to cadmium of human osteosarcoma cells is attributed, at least partly, to induction of metallothionein synthesis by cadmium and zinc in this cell line; in the rat osteosarcoma cell line, they do not induce metallothionein synthesis. Human osteosarcoma cells were more sensitive than rat osteosarcoma cells to lead with inhibition (IC50) of proliferation at 4 mol l lead and cytotoxicity at 20 versus 6 and over 20 moll lead, respectively, for these variables in rat osteosarcoma cells. Both cells lines attained the highest lead concentration in the 15 000 × g (mitochondrial) fraction. The lead in the mitochondrial, microsomal, nuclear and cytosolic fractions of the human cell line did not decrease during 24 h post-washout. Binding of lead was much less stable in the less sensitive rat cells, with 50–100% loss of mitochondrial, microsomal and nuclear lead during 24 h post-washout.  相似文献   

11.
Lactulose production from lactose and fructose was investigated with several commercial -galactosidases. The enzyme from Kluyveromyces lactis exhibited the highest lactulose productivity among the -galactosidases tested. The reaction conditions for lactulose production were optimized using cells that had been permeabilized by treatment with 50% (v/v) ethanol: cell concentration, 10.4 g l–1; concentration of substrates, 40% (w/v) lactose and 20% (w/v) fructose; temperature, 60°C; pH 7.0. Under these conditions, the permeabilized cells produced approximately 20 g l–1 lactulose in 3 h with a lactulose productivity of 6.8 g l–1 h–1. These results represent 1.3- and 2.1-fold increases in lactulose concentration and productivity compared with untreated washed cells. This is the first reported trial of enzymatic synthesis of lactulose using permeabilized yeast cells.  相似文献   

12.
Lee SO  Kim CS  Cho SK  Choi HJ  Ji GE  Oh DK 《Biotechnology letters》2003,25(12):935-938
Conjugated linoleic acid (CLA) was produced at 300 mg l–1 after 24 h culture of Lactobacillus reuteri in de Man–Rogosa–Sharpe medium containing 0.9 g linoleic acid (LA) l–1 and 1.67% (v/v) Tween 80. CLA was mainly located in the extracellular space of the cells. Washed cells previously grown on LA were less active than unadapted washed cells in converting LA into CLA. Most of the CLA transformed by washed L. reuteri cells was located in cells or associated with cells. CLA production by washed L. reuteri cells was most efficient in conversion with 0.45 g LA l–1 at pH 9.5 and 37°C for 1 h.  相似文献   

13.
Biomass and eicosapentaenoic acid (EPA) productivities were investigated in a flat panel airlift loop reactor ideally mixed by static mixers. Growth with ammonium, urea and nitrate as nitrogen source were performed at different aeration rates. Cultures grew on ammonium but the decay of pH strongly inhibited biomass increase. On urea biomass productivity reached 2.35 g L–1d–1at an aeration rate of 0.66 vvm (24 h light per day, 1000 mol photon m–2s–1). Aeration rates between 0.33 vvm and 0.66 vvm and maximal productivities on urea were linearly dependent. Productivity on nitrate never exceeded 1.37 g L–1d–1. In the range of maximum productivity photosynthesis efficiency of 10.6% was reached at low irradiance (250 mol photon m–2s–1). Photosynthesis efficiency decreased to 4.8% at 1000 mol photon m–2s–1. At these high irradiances the flat panel airlift reactor showed a 35% higher volume productivity than the bubble column. At continuous culture conditions the influence of CO2concentration in the supply air was tested. Highest productivities were reached at 1.25% (v/v) CO2where the continuous culture yielded 1.04 g L–1d–1(16 h light per day, 1000 mol photon m–2s–1). The average EPA content amounted to 5.0% of cell dry weight, that resulted in EPA productivities of 52 mg L–1d–1(continuous culture, 16 h light per day) or 118 mg L–1d–1(batch culture, 24 h light per day).  相似文献   

14.
Cummings  E.  Hundal  H.S.  Wackerhage  H.  Hope  M.  Belle  M.  Adeghate  E.  Singh  J. 《Molecular and cellular biochemistry》2004,261(1):99-104
The fruit of Momordica charantia (family: Cucurbitacea) is used widely as a hypoglycaemic agent to treat diabetes mellitus (DM). The mechanism of the hypoglycaemic action of M. charantia in vitro is not fully understood. This study investigated the effect of M. charantia juice on either 3H-2-deoxyglucose or N-methyl-amino-a-isobutyric acid (14C-Me-AIB) uptake in L6 rat muscle cells cultured to the myotube stage. The fresh juice was centrifuged at 5000 rpm and the supernatant lyophilised. L6 myotubes were incubated with either insulin (100 nM), different concentrations (1–10 g ml–1) of the juice or its chloroform extract or wortmannin (100 nM) over a period of 1–6 h. The results were expressed as pmol min–1 (mg cell protein)–1, n= 6–8 for each value. Basal 3H-deoxyglucose and 14C-Me-AIB uptakes by L6 myotubes after 1 h of incubation were (means ± S.E.M.) 32.14 ± 1.34 and 13.48 ± 1.86 pmol min–1 (mg cell protein)–1, respectively. Incubation of L6 myotubes with 100 nM insulin for 1 h resulted in significant (ANOVA, p < 0.05) increases in 3H-deoxyglucose and 14C-Me-AIB uptakes. Typically, 3H-deoxyglucose and 14C-Me-AIB uptakes in the presence of insulin were 58.57 ± 4.49 and 29.52 ± 3.41 pmol min–1 (mg cell protein–1), respectively. Incubation of L6 myotubes with three different concentrations (1, 5 and 10 g ml–1) of either the lyophilised juice or its chloroform extract resulted in time-dependent increases in 3H-deoxy-D-glucose and 14C-Me-AIB uptakes, with maximal uptakes occurring at a concentration of 5 g ml–1. Incubation of either insulin or the juice in the presence of wortmannin (a phosphatidylinositol 3-kinase inhibitor) resulted in a marked inhibition of 3H-deoxyglucose by L6 myotubes compared to the uptake obtained with either insulin or the juice alone. The results indicate that M. charantia fruit juice acts like insulin to exert its hypoglycaemic effect and moreover, it can stimulate amino acid uptake into skeletal muscle cells just like insulin. (Mol Cell Biochem 261: 99–104, 2004)  相似文献   

15.
This study examines the influence of current velocity in the toxiceffect of copper in diatom-dominated biofilms grown in artificial channels.Effects on community structure, algal biomass and photosynthesis (carbonincorporation) caused by 15 g L–1 of copperwere tested at contrasting (1 and 15 cm s–1)velocities. Moreover, a possible threshold on the effect of copper on algalbiomass and photosynthesis related to current velocity was examined by usingprogressively increasing current velocity (1 to 50 cms–1) at 15 g L–1 Cu.Chlorophyll-a decreased ca. 50% as a result of addition of15 g L–1 Cu. Chlorophyll decrease occurredearlier at 15 cm s–1 than at 1 cms–1 when adding 15 g L–1Cu. Copper also caused a remarkable decrease in carbon incorporation(from 30 to ca. 50%), which was produced earlier at 15 cms–1 (three days) than at 1 cms–1 (seven days). Some taxa were affected by thecombination of copper and current velocity. Both Achnanthesminutissima and Stigeoclonium tenue becomedominant at 15 cm s–1 in the presence of copper.Significant inhibition of algal growth in 15 g L–1Cu occurred at low (1 cm s–1) and highvelocities (50 cm s–1), but not at intermediatevelocity (20 cm s–1). The experiments indicatethat current velocity triggers the effect that copper has on diatom-dominatedbiofilms, and that the effect is more remarkable at low and high than atintermediate current velocities.  相似文献   

16.
Whole cells, cytoplasms and peptidoglycans of ten different lactic acid bacteria (LAB) were tested for in vitro cytotoxicity on diverse cancer cell lines using the 3H-thymidine incorporation assay. The peptidoglycans and cytoplasm fractions, as well as heat-killed whole cells of LAB, had significant antiproliferative activities against several cancer cell lines. In particular, the cytoplasm fractions exhibited marked direct antiproliferative activities against colon and gastric cancer cell lines, whereas the peptidoglycans retarded growth of colon and bladder cancer cell lines. The cytoplasm fractions of Bifidobacterium longum and Lactococcus lactis ssp. lactis inhibited proliferation of two cancer cell lines by 50% at 33 and 23 g ml–1 for SNUC2A (a human colon adenocarcinoma cell line) and 17 and 11 g ml–1 for SNU-1 (a human gastric cancer cell line), respectively.  相似文献   

17.
Summary Intact and osmotically sensitive cells of Corynebacterium glutamicum can be efficiently transformed by electroporation. This was shown by using the plasmid vector pUL-330 (5.2 kb), containing the kanamycin resistance gene of transposon Tn5. The following electric parameters yielded efficient transformation. For intact cells: one exponentially decaying field pulse with time constants and with initial field intensities of E 0=35–40 kV cm-1; prepulse temperature 20°C. Cell regeneration (survival) was 100%–80%. Transformation efficiency can be increased by an additional freeze and thaw cycle of the cells, prior to electroporation. Lysozyme treated cells (osmotically sensitive) were transformed with three successive pulses of E 0=25–30 kV cm-1. Cell regeneration under these conditions was found to be 20–30%. The optimum yield of transformants/g plasmid-DNA was 3×103 for intact cells, 2×104 for intact cells which were frozen and thawed twice and 7×104 for osmotically sensitive cells if the cell suspension was pulsed at a cell density of 1–3×108/ml and at a DNA concentration of 0.2 g/ml up to 2 g/ml. The data obtained for osmotically sensitive cells suggest that the temperature increase accompanying the electric field pulse enhances colony formation and transformation efficiency if the initial prepulse temperature is 20°C, although regeneration of electroporated C. glutamicum cells starts to decrease at temperatures20°C.  相似文献   

18.
Summary Incubation of human lymphocytes with recombinant interleukin-2 (rIL-2) results in the generation of lymphokine-activated killer (LAK) cells capable of lysing a wide variety of tumor cells. The present study was undertaken to examine the effect of recombinant interferon (rIFN-) on LAK cell cytotoxicity generated from different peripheral blood mononuclear cell (PBMC) subpopulations. When unseparated PBMC were stimulated by rIL-2 and rIFN-, the latter induced a transient enhancement after 2 days followed by a suppression of LAK cell cytotoxicity at day 6. Enhancement of LAK cell cytotoxicity was moderate and inconstant, whereas the inhibition was strong and observed with all the donors tested. This suppression was not associated with a decrease in the [3H]thymidine uptake. PBMC depleted of adherent cells were more sensitive to the stimulation by rIL-2 and the induced cytotoxicity was not modified by rIFN-. Monocyte-enriched plastic-adherent cells, when incubated with rIL-2 and rIFN-, became cytotoxic after 2–3 days of culture and inhibited LAK cell activity after 5–6 days. Collectively, our results suggest that rIFN- affects LAK cell cytotoxicity through the activation of plastic-adherent, monocyte-rich, cells which modulate natural killer cells, first in a positive, then in a negative way.  相似文献   

19.
Summary Dantrolene-Na is a muscular relaxant which binds to sarcoplasmic reticulum (SR) with high affinity and decreases the availability of Ca2+ channels. The binding of fluorescent compounds, dantrolene-Na, nifedipine and chlortetracycline to the ciliary membrane ofParamecium aurelia has been studied. Dantrolene at the concentrations of 1.9 · 10–5, 3.8 · 10–5 and 7.9 · 10–5 M manifested a punctuated binding pattern to the cell membrane. Isolated cilia also bound dantrolene at their basal portion, whereas deciliated cell bodies lost their dotted binding pattern. Chlortetracycline showed a similar but weaker fluorescent staining. Nifedipine treated cells revealed no sign of fluorescent binding to the membrane and was only taken up in food vacuoles.Based on these observations we propose that dantrolene binding regular arrays ofParamecium cell membrane could be identical to granular plaques observed by electron microscope. The possible functioning of these structures as Ca2+ reservoirs is also discussed.  相似文献   

20.
Summary The ethanol yield was not affected and the ethanol productivity increased (10%) when linearly decreasing feeding rates were used instead of constant feeding rates in fed-batch ethanol fermentations.Nomenclature F reactor feeding rate (L.h–1) - ME mass of ethanol in the fermentor (g) - Ms mass of TRS in the fermentor (g) - Mx mass of yeast cells (dry matter) in the fermentor (g) - P ethanol productivity (g.L–1.h–1) - s standard deviation - So TRS concentration in the feeding mash (g.L–1) - t time (h) - T fermentor filling-up time (h) - TRS total reducing sugars calculated as glucose (g.L–1) - Xo yeast cells concentration (dry matter) in the inoculum (g.L–1) - average ethanol yield (% of the theoretical value)  相似文献   

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