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1.
Katrin Spiesberger Florian Paulfranz Anton Egger Judith Reiser Claus Vogl Judith Rudolf-Scholik Corina Mayrhofer Ludger Grosse-Hovest Gottfried Brem 《PloS one》2015,10(10)
Background
30 years ago, the potential of bispecific antibodies to engage cytotoxic T cells for the lysis of cancer cells was discovered. Today a variety of bispecific antibodies against diverse cell surface structures have been developed, the majority of them produced in mammalian cell culture systems. Beside the r28M, described here, no such bispecific antibody is known to be expressed by transgenic livestock, although various biologicals for medical needs are already harvested—mostly from the milk—of these transgenics. In this study we investigated the large-scale purification and biological activity of the bispecific antibody r28M, expressed in the blood of transgenic cattle. This tandem single-chain variable fragment antibody is designed to target human CD28 and the melanoma/glioblastoma-associated cell surface chondroitin sulfate proteoglycan 4 (CSPG4).Results
With the described optimized purification protocol an average yield of 30 mg enriched r28M fraction out of 2 liters bovine plasma could be obtained. Separation of this enriched fraction by size exclusion chromatography into monomers, dimers and aggregates and further testing regarding the biological activity revealed the monomer fraction as being the most appropriate one to continue working with. The detailed characterization of the antibody’s activity confirmed its high specificity to induce the killing of CSPG4 positive cells. In addition, first insights into tumor cell death pathways mediated by r28M-activated peripheral blood mononuclear cells were gained. In consideration of possible applications in vivo we also tested the effect of the addition of different excipients to r28M.Conclusion
Summing up, we managed to purify monomeric r28M from bovine plasma in a large-scale preparation and could prove that its biological activity is unaffected and still highly specific and thus, might be applicable for the treatment of melanoma. 相似文献2.
Milica D olovi GM Jankovi RB olovi Vesna M Martinovi-emeriki 《Cancer immunology, immunotherapy : CII》1998,15(4):286-288
A case of primary nonsecretory plasmacytoma of the spleen is reported. On laparotomy and splenectomy a 920 g spleen was removed,
measuring 16×14×6 cm. The cut surface of the entire spleen showed that the tumour occupied most of the splenic tissue. A bone
marrow aspirate and trephine, skeletal survey showed no signs of myeloma. Biopsy of the liver and regional lymph nodes was
normal. Immunocytochemistry of the splenic tumour showed positivity for pan-B and plasma cell markers. After splenectomy the
patient was treated with chemotherapy according to protocol VBCMP (M2). 相似文献
3.
4.
Zev A. Binder Kelli M. Wilson Vafi Salmasi Brent A. Orr Charles G. Eberhart I-Mei Siu Michael Lim Jon D. Weingart Alfredo Quinones-Hinojosa Chetan Bettegowda Amin B. Kassam Alessandro Olivi Henry Brem Gregory J. Riggins Gary L. Gallia 《PloS one》2016,11(3)
Objective
Human tumor cell lines form the basis of the majority of present day laboratory cancer research. These models are vital to studying the molecular biology of tumors and preclinical testing of new therapies. When compared to traditional adherent cell lines, suspension cell lines recapitulate the genetic profiles and histologic features of glioblastoma multiforme (GBM) with higher fidelity. Using a modified neural stem cell culture technique, here we report the characterization of GBM cell lines including GBM variants.Methods
Tumor tissue samples were obtained intra-operatively and cultured in neural stem cell conditions containing growth factors. Tumor lines were characterized in vitro using differentiation assays followed by immunostaining for lineage-specific markers. In vivo tumor formation was assayed by orthotopic injection in nude mice. Genetic uniqueness was confirmed via short tandem repeat (STR) DNA profiling.Results
Thirteen oncosphere lines derived from GBM and GBM variants, including a GBM with PNET features and a GBM with oligodendroglioma component, were established. All unique lines showed distinct genetic profiles by STR profiling. The lines assayed demonstrated a range of in vitro growth rates. Multipotency was confirmed using in vitro differentiation. Tumor formation demonstrated histologic features consistent with high grade gliomas, including invasion, necrosis, abnormal vascularization, and high mitotic rate. Xenografts derived from the GBM variants maintained histopathological features of the primary tumors.Conclusions
We have generated and characterized GBM suspension lines derived from patients with GBMs and GBM variants. These oncosphere cell lines will expand the resources available for preclinical study. 相似文献5.
6.
We studied electrofusion of mouse two-cell embryos in order to define parameters which would result in a high yield of fused embryos. Various cell alignment times (from <10 to >60 s) and alternating current percentages (2 to 100%) were examined. The fusion parameters tested were the number of fusion pulses (1-9), pulse length (30-90 mus) and pulse strength (0.50-1.79 kV/cm). Furthermore different combinations of these three parameters were tested. In addition the influence of several embryo culture media on the fusion rates was examined. The results show that the fusion rate of the embryos increases with shorter alignment and higher percentages of the alternating current. The highest fusion rate (95%) was obtained by use of one pulse with a duration of 70 mus and a field strength of 0.60-0.79 kV/cm. The survival rate of the embryos was best if Whitten Medium was used before and after the fusion pulses. The fusion of two-cell stages results in tetraploid embryos which can serve as models for studies in polyploid cells. 相似文献
7.
8.
Jürgen Brem Mihaela C. TurcuCsaba Paizs Katri LundellMonica-Ioana To?a Florin-Dan IrimieLiisa T. Kanerva 《Process Biochemistry》2012,47(1):119-126
Lipase AK “Amano” 20 from Pseudomonas fluorescens (PFL) was immobilized using diverse immobilization techniques. The methods developed, especially the optimized sol-gel procedure, enabled the fine tuning of enzymatic activity and enantioselectivity for the kinetic resolution of racemic ethyl 3-aryl-3-hydroxypropanoates. The aryl moieties of the racemates include furan-2 and 3-yl, thiophen-2 and 3-yl, benzofuran-2-yl, benzo[b]thiophen-2-yl, as well as phenyl and 4-chloro- and 4-methoxyphenyl groups. The optimized PFL sol-gel preparation (encapsulation from the aqueous solution of PFL, sucrose and Celite in situ) was shown to be efficiently reusable in ten cycles and highly enantioselective with E > 200 to all other substrates except furan-2 and 3-yl and thiophen-2 and 3-yl substituted compounds with E 108-184. 相似文献
9.
Irp9, encoded by the high-pathogenicity island of Yersinia enterocolitica,is able to convert chorismate into salicylate,the precursor of the siderophore yersiniabactin
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The Irp9 protein of Yersinia enterocolitica participates in the synthesis of salicylate, the precursor of the siderophore yersiniabactin. In Pseudomonas species, salicylate synthesis is mediated by two enzymes: isochorismate synthase and isochorismate pyruvate-lyase. Both enzymes are required for complementation of a Yersinia irp9 mutant. However, irp9 is not able to complement Escherichia coli entC for the production of enterobactin, which requires isochorismate as a precursor. These results suggest that Irp9 directly converts chorismate into salicylate. 相似文献
10.
Intraspecific competition of endophyte infected vs uninfected plants of two woodland grass species 总被引:3,自引:0,他引:3
Grass endophytes (Clavicipitaceae, Ascomycota) are generally considered to be mutualists which increase the host's fitness. Infected plants are often more persistent and competitive than uninfected plants, influencing population dynamics and plant community diversity. However, most of this empirical evidence is based on studies focusing on agronomically important grass species such as tall fescue or perennial ryegrass and their implications for livestock and man-made habitats. Recent studies indicate that endophyte-plant associations may be more variable, ranging from parasitic to mutualistic. In the present study, we investigated the influence of endophyte infection on two wild woodland grasses, which are naturally infected with distinct fungal endophytes: Brachypodium sylvaticum with Epichloë sylvatica and Bromus benekenii with Epichloë bromicola . An intraspecific competition experiment was conducted over two growing seasons in the greenhouse and in an experimental garden. At first harvest (after 12 weeks growing), endophyte infection had a significant negative effect on above ground dry matter yield (DMY) of B. sylvaticum , but a significant positive effect on DMY of Br. benekenii under competition. The same differential effects on DMY and on total seed number were also observed at final harvest (after 62 weeks growing). Results from Br. benekenii were consistent with our hypothesis of increased competitive abilities of infected plants in nature which could explain the high infection rate observed in natural populations. In contrast, this explanation does not hold true for B. sylvaticum , and other factors such as increased herbivore and pathogen resistance together with frequent horizontal transmission may be responsible for the very high incidence of this association in nature. Our results confirm previous predictions that beneficial effects of endophyte infection in wild grasses can vary for different grass species, even in comparable habitats. 相似文献