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81.
Lindl T 《Cytotechnology》1996,21(3):183-193
This article describes the current status in the development of human monoclonal antibodies. Over the last ten years a lot of information about the human immune system has emerged. Combining these with the many new (bio-)technologies it is plausible that the long awaited breakthrough of this technology is close. This paper focuses on the classical cell-biological methods of achieving stable, antibody-producing human cell lines via cell fusion methods or virus derived transformations of human B-lymphocytes, as well as genetic engineering methods e.g. DNA libraries or phage display technology. The available in vitro immunization methods are critically reviewed and their impact on this topic is discussed. Therapeutic applications for cancer treatment or passive immunization against infectious diseases with antibodies derived by both ways are also reviewed. 相似文献
82.
The antitumor effect of the combined administration with recombinant human interleukin-2 (rIL-2) and sizofiran (SPG), a single
glucan of Shizophyllum commune Fries, was studied in vivo in C57BL/6 mice intraperitoneally inoculated with EL-4 lymphoma.
The effect was evaluated by a) comparing the survival time of the mice, b) analysis of the intraperitoneal cell population
in Giemsa-stained specimens, c) surface marker analysis of peritoneal exudative cells with flow cytometry, d) cytotoxic assay
of cells against EL-4 and Yac-1 lymphoma, and e) elimination of some cell populations by monoclonal antibodies, to identify
the antitumor-effector cells showing cytotoxic activity. The survival of mice given both rIL-2 and SPG was significantly longer
than the control mice or those given SPG alone or rIL-2 alone. It was demonstrated that the administration of SPG and/or rIL-2
to the EL-4 lymphoma-bearing mice activated immune-response cells in the peritoneal cavity such as T lymphocytes, NK cells,
or macrophages, which might be effective in reducing lymphoma cells. The combination of rIL-2 and SPG administration appears
to activate the antitumor- immune response at the tumor site more effectively than when either agent was administered alone. 相似文献
83.
Dirk Schories 《Aquatic Ecology》1995,29(3-4):341-347
For the last two decades dense mats of species of the filamentous green algaeEnteromorpha spp. have regulary occurred on tidal flats of Köningshafen Bay (island Sylt, North Sea, FRG). In calm areas overwintering of adult plants or plant fragments is a common process to guarantee the mass development during the next season. In contrast, the distribution ofEnteromorpha on exposed sandy tidal flats depends on recruitment by juvenile stages. In 1993Enteromorpha spore settlement was recorded regularly in the field. Polyethylene dishes were placed in the field and left for a period of seven days and lateron cultivated in the laboratory to checkEnteromorpha germling development. During summer 1993 — at a minimum distance of 200 m to the nearest adultEnteromorpha populations — a total of at least 82×106 spores m–2 settled. During winter the number of spores attached to the collecting dishes was close to zero and the adjacent sand flats were free of any visibleEnteromorpha plants. In further experiments it was shown that the development ofEnteromorpha juveniles in the next spring depended on the overwintering capacity of spores. More than 2×106 spores m–2 attached to large sand grains and other substrata (e.g. Hydrobia ulvae) survived the winter. In a laboratory experiment several species ofEnteromorpha were able to survive in total darkness for at least 10 months. 相似文献
84.
Roberto E. Izquierdo Kimberly Breese Shalini Jain Daniel Carestio Lawrence Jung James Figge 《In vitro cellular & developmental biology. Animal》1995,31(1):71-76
Summary Gene transfer techniques can be used to encode the production of a polypeptide product, such as human growth hormone (hGH),
that is missing in an acquired or inherited disease state such as growth hormone deficiency. In one model system, engineered
C2C12 myoblasts are injected intramuscularly into a mouse and subsequently secrete hGH into the circulation. In this regard,
a gene-expression regulatory system that functions in myoblasts would be of interest. We demonstrate that theEscherichia coli lac operon system can be used to stringently regulate the expression of hGH in engineered C2C12 myoblasts in tissue culture.
A DNA segment encoding hGH was linked to a DNA segment containing an SV40 enhancer and promoter. The latter components were
positioned between two syntheticlac operators.Lac repressor expression was driven by a simian cytomegalovirus promoter. In transient co-transfection assays, hGH expression
from cultured C2C12 myoblasts could be modulated up to 60-fold (P = 0.002) with the inducing agent, isopropyl-β-d-thiogalactoside (IPTG). In the absence of IPTG, hGH expression was almost fully repressed. These results show that the components
of theE. coli lac operon provide a stringent regulatory system for use in myoblasts. The system might prove to be useful for the regulation
of transferred genes in animals. 相似文献
85.
Encephalitozoon cuniculi Isolated from the Urine of an AIDS Patient, which Differs from Canine and Murine Isolates 总被引:4,自引:0,他引:4
WAFAA S. HOLLISTER ELIZABETH U. CANNING NICOLA I. COLBOURN EMMA J. AARONS 《The Journal of eukaryotic microbiology》1995,42(4):367-372
ABSTRACT. A species of Encephalitozoon has been isolated from the urine of a patient with the acquired immunodeficiency syndrome and maintained in vitro in Madin Darby Canine Kidney cells. When examined by random amplified polymoprhic DNA polymerase chain reaction the new isolate was found to differ from E. hellem and to have amplified products in common with murine and canine E. cuniculi . However, it more closely resembled the canine than the murine isolate. Sodium dodecylsulphate polyacrylamide gel electrophoresis differentiated between all three isolates of E. cuniculi , with a band at 42–45 kDa present in the murine isolate only, bands at 52 kDa present in the canine and human isolates but not the murine, and a single band at 60 kDa (murine) and 65 kDa (canine) replaced by two bands at 55 and 70 kDa in the human isolate. The 55 kDa and 70 kDa antigens were also revealed as characteristic bands of the human isolate by Western blotting. The study has thus revealed that the species Encephalitozoon cuniculi is not a homogeneous entity. 相似文献
86.
87.
Angelo De Milito Marinunzia Catucci Francesco Iannelli Laura Romano Maurizio Zazzi Pier Egisto Valensin 《Molecular biotechnology》1995,3(2):166-169
A reliable selective PCR procedure that combines the use of additionally mutated primers with the specificity-enhancing properties
of a commercial preparation (Perfect Match, Stratagene) is described. The human immunodeficiency virus type 1pol gene point mutations known to confer in vitro resistance to azidothymidine were examined as a model for optimization of the
assay. The usual strategy of deliberately introducing an additional mismatch 1 residue from the 3′ end in the wild-type and
mutant primers did not allow reproducible discrimination between wild-type and mutant target sequences. Addition of minimal
amounts of Perfect Match to the same PCR mixtures resulted in a significantly enlarged range of selective annealing temperatures,
providing a valuable and cost-effective means for reliable detection of known mutations by selectivePCR. 相似文献
88.
Rubén C. Aguilar Lilia A. Retegui Marie-Catherine Postel-Vinay Leonor P. Roguin 《Molecular and cellular biochemistry》1994,136(1):35-42
We have previously shown that a monoclonal antibody (MAb) recognizing the human growth hormone (hGH) antigenic domain left exposed after binding to lactogenic receptors enhanced hGH binding probably through allosteric effects on the hormone binding site. Since receptors displaying different specificities would not recognize exactly the same hGH region, we explored whether some of our MAb could affect hGH binding to somatogenic receptors from rabbit liver and to human liver hGH-specific receptors.The effect of MAbAE5, AC8 and F11 on hGH binding was measured by determining the formation of125I-MAb:hGH:receptor complexes using two different experimental approaches. Results from procedure A, which involved the previous binding of the hormone to microsomes before adding125I-MAb, indicated that the hGH domain defined by epitopes AE5, AC8 and F11 is uncovered in the various hormone:receptor complexes.Procedure B was devised to reveal any alteration in the hGH molecule induced by the MAb. In this case preformed125I-MAb:hGH complexes were added to microsomes. Data showed that125I-MAb AE5:hGH complexes bound better to the various receptors than125I-MAb AE5 to hGH:receptor complexes. On the contrary, hGH previously bound to125I-MAb AC8 or125I-MAb F11 was less recognized by the receptors than the free hormone. Furthermore, binding of MAb AE5 or MAb F11 to hGH 20 K (a natural hGH variant lacking residues 32–46) also enhanced its affinity to the various receptors whereas MAb AC8 did not inhibit hGH 20 K binding.Results indicated that MAb recognizing the hGH antigenic area that remains unmasked after binding to different membrane-bound receptors are able to affect hormone binding site. MAb would induce either positive or negative allosteric changes in the hormone region involved in its binding to lactogenic, somatogenic and hGH-specific receptors. 相似文献
89.
Prakash Sista Sharon Edmiston James W. Darges Simon Robinson David J. Burns 《Molecular and cellular biochemistry》1994,141(2):129-134
Transmission of extra cellular signals across biological membranes results in the generation of lipid metabolites which in turn influence specific cellular events such as cell growth or differentiation. Many of these lipid messengers can activate protein kinase C (PKC) isozymes of which one function is to perpetuate the extracellular signals to the nucleus by phosphorylating other targets proteins. We have engineered mammalian cell lines to identify and evaluate activators and inhibitors of PKC-dependent and independent signal transduction pathways. The A31 mouse fibroblast cell line, has been stably transfected with a construct containing a triplet repeat of the TPA response element (TRE) upstream of a thymidine kinase promoter fused to the human growth hormone (hGH) gene. A31 cells containing this reporter construct exhibit significant increases in hGH secretion following stimulation by phorbol esters or other mitogens. The levels of hGH secretion are modulated in this system using different pharmacological agents. We demonstrate that this assay can be used to identify specific and general inhibitors as well as activators of the signal transduction pathway mediated by PKC isozymes. (Mol Cell Biochem141: 129–134, 1994) 相似文献
90.
Michael A. Rogers Jürgen Schweizer Thomas Krieg Hermelita Winter 《Molecular biology reports》1994,20(3):155-161
We present the nucleotide and amino acid sequence for a novel human type I hair keratin, which could be identified through its high sequence homology and strict carboxyterminal length identity as a human ortholog of the murine hair keratin mHa3. Our hHa3 sequence differs, however, from that of a previously described hHa3 hair keratin (published only as an amino acid sequence; [13]) in 24 amino acid positions, 8 of which occur in the middle of the carboxyterminal domain. PCR of genomic DNA from 25 normal human subjects using a primer pair derived from sequence segments located in the 3-region of our hHa3 clone that encode conserved amino acid sequences in both keratins, resulted in the amplification of two distinct products of 0.38 kbp and 1.0 kbp. DNA sequence analysis of the cloned PCR products allowed identification of the 0.38 kb sequence as that originating from Yuet al. [13] and the 1.0 kb sequence as that being derived from our data. The difference in fragment length was due to unique intron 6 sequences, indicating that these two keratin species are encoded by genes of their own. Moreover, extensive Southern blot analyses with DNA from 25 unrelated individuals of different races using a 3-noncoding sequence from our keratin and the intron 6 sequence of the keratin of Yuet al. [13], as hybridization probes showed that both keratin genes are present as single copy sequences occurring ubiquitously and without gross alterations in the human genome. Collectively, these data demonstrate that the human type I hair keratin described in this paper represents an isoform of the previously described hHa3 keratin. We propose that these hHa3 isoforms be named in chronological order of discovery hHa3-I and hHa3-II. 相似文献