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1.
2.
Peter J. K. Kuppen Alexander M. M. Eggermont Andreas Marinelli Emile de Heer Cornelis J. H. van de Velde Gert Jan Fleuren 《Cancer immunology, immunotherapy : CII》1991,33(1):28-32
Summary The role of 2-mercaptoethanol and indomethacin in the induction of lymphokine-activated killer (LAK) activity by interleukin-2 (IL-2) in rat splenocyte cultures was investigated. Spleens from 4-month-old male rats of five different strains were tested. Splenocytes were cultured for 3–5 days in the presence of IL-2 (1000 U/ml) and LAK activity was assessed by 4-h51Cr release assays with P815 and YAC-1 cells as targets. LAK activity could be induced by IL-2 in splenocytes from all rat strains, but only when 2-mercaptoethanol was present in the culture medium. Optimal LAK activity was induced when the 2-mercaptoethanol concentration in splenocyte cultures was at least 5 µM. Different rat strains showed differences in levels of in vitro induction of LAK activity. In the presence of 2-mercaptoethanol the level of LAK activity induced by IL-2 was high in BN and Lewis rats, intermediate in Wistar and Wag rats, and low in DZB rats. In the absence of 2-mercaptoethanol no or minimal LAK activity was induced. Furthermore we observed that addition of 50 µm indomethacin to the culture medium in the presence of 2-mercaptoethanol augmented the induction of LAK activity to some extent. In the absence of 2-mercaptoethanol, addition of indomethacin resulted only in low levels or no induction of LAK activity. We conclude that for optimal induction of LAK activity by IL-2 in rat splenocyte cultures 2-mercaptoethanol is essential, while indomethacin can only marginally further improve this induction. 相似文献
3.
Alexander M. Eggermont Willem Weimar Bhupendra Tank Amelie M. Dekkers-Bijma Richard L. Marquet Johannes S. Lameris Dick L. Westbroek Johannes Jeekel 《Cancer immunology, immunotherapy : CII》1986,21(1):81-84
Summary A total of 20 patients with advanced colorectal cancer received recombinant leukocyte interferon-A (rIFNA) either chronically (group I: twice a week up to 20×106 IU/m2 i.m.) or cyclically (group II: 1–4 periods of 8 consecutive days up to 20×106 IU/m2 i.m. daily at 20-days intervals) over a period of 12 weeks. There was 1 partial response, 1 mixed response and 1 patient with stable disease, whilst 17 patients had progressive disease. Median survival was 15.5 months. Survival was significantly shorter when the extent of hepatic disease was >25% (P=0.05), extrahepatic disease was extensive (P<0.005), alkaline phosphatase level was >2× normal (P<0.02), or performance status was <100% (P<0.001). Toxicity consisting mainly of fever, fatigue, anorexia and weight loss was serious in group I and minimal in group II. Administration of rIFNA led to a short lived augmentation of natural killer (NK) cell activity. In the cyclically treated group this was a recurrent phenomenon whereas a marked lasting depression of NK cell activity was seen in chronically treated patients. Interferon- production capacity was significantly stimulated during rIFNA therapy. The differences in toxicity and immunostimulatory effects between the two schedules may be of importance in the design of further studies.This trial was supported in part by Hoffmann-La Roche, Basle 相似文献
4.
An De Bondt Kristel Eggermont Iris Penninckx Inge Goderis Willem F. Broekaert 《Plant cell reports》1996,15(7):549-554
We have previously developed a protocol for efficient gene transfer and regeneration of transgenic calli following cocultivation of apple (cv. Jonagold) explants with Agrobacterium tumefaciens (De Bondt et al. 1994, Plant Cell Reports 13: 587–593). Now we report on the optimization of postcultivation conditions for efficient and reproducible regeneration of transgenic shoots from the apple cultivar Jonagold. Factors which were found to be essential for efficient shoot regeneration were the use of gelrite as a gelling agent and the use of the cytokinin-mimicing thidiazuron in the selective postcultivation medium. Improved transformation efficiencies were obtained by combining the hormones thidiazuron and zeatin and by using leaf explants from in vitro grown shoots not older than 4 weeks after multiplication. Attempts to use phosphinothricin acetyl transferase as a selectable marker were not successful. Using selection on kanamycin under optimal postcultivation conditions, about 2% of the leaf explants developed transgenic shoots or shoot clusters. The presence and expression of the transferred genes was verified by -glucuronidase assays and Southern analysis. The transformation procedure has also been succesfully applied to several other apple cultivars.Abbreviations BAP
benzylaminopurine
- CTAB
hexadecyltrimethylammoniumbromide
- Na2EDTA
ethylenediamine-tetra-acetate ferric-sodium salt
- FeNaEDTA
ethylenediamine-tetra-acetate ferric-sodium salt
- GA3
gibberellic acid 3
- GusA
-glucuronidase
-
gusA
-glucuronidase gene of Escherichia coli
- IAA
indole acetic acid
- IBA
indole butyric acid
- 2iP
N6-2-isopentenyl adenine
- NAA
naphthalene acetic acid
-
nptII
neomycinphosphotransferase II gene
-
bar
phosphinothricin acetyl transferase gene
- PCR
polymerase chain reaction
- PPT
phosphinothricin
- STS
silver thiosulphate
- T-DNA
transferred DNA
- TDZ
thidiazuron
- X-Gluc
5-bromo-4-chloro-3-indolyl -D-glucuronide
- Zea
trans-Zeatin 相似文献
5.
Kristel Eggermont Inge J. Goderis Willem F. Broekaert 《Plant Molecular Biology Reporter》1996,14(3):273-279
We describe here a reliable high-throughput method for extraction of RNA from fresh or frozen plant tissue that obviates laborious
and time-consuming homogenisation by mortar and pestle. The method is based on homogenisation by high-speed reciprocal shaking
in presence of a mixture of inexpensive abrasive materials; i.e., quartz sand and glass beads. After homogenisation, the method
follows a standard procedure for RNA extraction by phenol/LiCl. Yield and quality of RNA obtained by homogenisation with the
sand/glass bead mix are identical to those obtained by mortar and pestle. 相似文献
6.
Wim Van de Vrie Sylke A. M. Van der Heyden Eric E. O. Gheuens Amelie M. Bijma Ernst A. De Bruijn Richard L. Marquet Allan T. Van Oosterom Alexander M. M. Eggermont 《Cancer immunology, immunotherapy : CII》1993,37(5):337-342
The development of resistance to anticancer drugs urges the search for different treatment modalities. Several investigators have reported the concomitant development of drug resistance and resistance to natural killer (NK), lymphokine-activated killer (LAK) or monocyte/macrophage cell lysis, while others described unchanged or even increased susceptibility. We investigated this subject in the rat colon carcinoma cell line, CC531-PAR, which is intrinsically multidrug-resistant (MDR), and in three sublines derived from this parental cell line: a cell line with an increased MDR phenotype (CC531-COL), a revertant line from CC531-COL (CC531-REV), which demonstrates enhanced sensitivity to anticancer drugs of the MDR phenotype, and an independently developed cisplatin-resistant line (CC531-CIS). In a 4-h51Cr-release assay we found no difference in susceptibility to NK cell lysis. No significant differences in lysability by adherent LAK (aLAK) cells were observed in a 4-h assay. In a prolonged 20-h51Cr-release assay an enhanced sensitivity to aLAK-cell-mediated lysis was observed in the revertant, P-glycoprotein-negative cell line and in the cisplatin-resistant cell line (CC531-CIS). None of the cell lines was completely resistant to lysis by aLAK cells. Therefore, a role for immunotherapy in the treatment of drug-resistant tumors remains a realistic option. 相似文献
7.
Pathogen-induced systemic activation of a plant defensin gene in Arabidopsis follows a salicylic acid-independent pathway. 总被引:21,自引:0,他引:21 下载免费PDF全文
I A Penninckx K Eggermont F R Terras B P Thomma G W De Samblanx A Buchala J P Mtraux J M Manners W F Broekaert 《The Plant cell》1996,8(12):2309-2323
A 5-kD plant defensin was purified from Arabidopsis leaves challenged with the fungus Alternaria brassicicola and shown to possess antifungal properties in vitro. The corresponding plant defensin gene was induced after treatment of leaves with methyl jasmonate or ethylene but not with salicylic acid or 2,6-dichloroisonicotinic acid. When challenged with A. brassicicola, the levels of the plant defensin protein and mRNA rose both in inoculated leaves and in nontreated leaves of inoculated plants (systemic leaves). These events coincided with an increase in the endogenous jasmonic acid content of both types of leaves. Systemic pathogen-induced expression of the plant defensin gene was unaffected in Arabidopsis transformants (nahG) or mutants (npr1 and cpr1) affected in the salicylic acid response but was strongly reduced in the Arabidopsis mutants eln2 and col1 that are blocked in their response to ethylene and methyl jasmonate, respectively. Our results indicate that systemic pathogen-induced expression of the plant defensin gene in Arabidopsis is independent of salicylic acid but requires components of the ethylene and jasmonic acid response. 相似文献
8.
J. P. Fryns M. Haspeslagh E. Vandenbussche P. Goddeeris E. Eggermont H. Van den Berghe 《Human genetics》1980,56(2):225-226
Summary An XX/XY mosaicism was detected in two phenotypically male newborns with perinatal death. The importance of chromosome studies at the paediatric necropsy and the significance of the present findings are discussed. 相似文献
9.
Tierens KF Thomma BP Bari RP Garmier M Eggermont K Brouwer M Penninckx IA Broekaert WF Cammue BP 《The Plant journal : for cell and molecular biology》2002,29(2):131-140
An Arabidopsis thaliana mutant, esa1, that shows enhanced susceptibility to the necrotrophic pathogens Alternaria brassicicola, Botrytis cinerea and Plectosphaerella cucumerina, but has wild-type levels of resistance to the biotrophic pathogens Pseudomonas syringae pv. tomato and Peronospora parasitica. The enhanced susceptibility towards necrotrophic pathogens correlated with a delayed induction of phytoalexin accumulation and delayed induction of the plant defensin gene PDF1.2 upon inoculation with pathogens. Two reactive oxygen generating compounds, paraquat and acifluorfen, were found to cause induction of both phytoalexin accumulation and PDF1.2 expression in wild-type plants, but this induction was almost completely abolished in esa1. This finding suggests that esa1 may somehow be involved in transduction of signals generated by reactive oxygen species. 相似文献