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Query: UMLS:C0272170 (SDS)
 50,377 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A plasmid pAc373GM-CSF was constructed and co-transfected into Spodoptera frugiperda (Sf9) cells with wild-type Autographa californica nuclear polyhedrosis virus (AcNPV) DNA. The recombinant virus vAc373GM-CSF was identified and purified by several rounds of plaque hybridization. By assaying the culture medium, we demonstrated recombinant virus infected Sf9 cells expressing hGM-CSF. Recombinant hGM-CSFs with apparent molecular masses of 14.5, 15.5 and 16.5 kDa were detected by the Western blot method. All 3 forms have biological activity of hGM-CSF. Following N-glycanase treatment, a single band of 14.5-15.5 kDa appeared in SDS-PAGE. Western blot analysis of expression in Sf9 cell treated with tunicamycin revealed only the presence of the 14.5 kDa species. Thus, the signal sequence of recombinant hGM-CSF could be recognized and cleaved by infected insect cell and the resultant molecule secreted into the media.
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PMID:Expression of human granulocyte-macrophage colony-stimulating factor gene in insect cells by a baculovirus vector. 240 25

A panel of antigen-specific mouse helper T cell clones was characterized according to patterns of lymphokine activity production, and two types of T cell were distinguished. Type 1 T helper cells (TH1) produced IL 2, interferon-gamma, GM-CSF, and IL 3 in response to antigen + presenting cells or to Con A, whereas type 2 helper T cells (TH2) produced IL 3, BSF1, and two other activities unique to the TH2 subset, a mast cell growth factor distinct from IL 3 and a T cell growth factor distinct from IL 2. Clones representing each type of T cell were characterized, and the pattern of lymphokine activities was consistent within each set. The secreted proteins induced by Con A were analyzed by biosynthetic labeling and SDS gel electrophoresis, and significant differences were seen between the two groups of T cell line. Both types of T cell grew in response to alternating cycles of antigen stimulation, followed by growth in IL 2-containing medium. Examples of both types of T cell were also specific for or restricted by the I region of the MHC, and the surface marker phenotype of the majority of both types was Ly-1+, Lyt-2-, L3T4+, Both types of helper T cell could provide help for B cells, but the nature of the help differed. TH1 cells were found among examples of T cell clones specific for chicken RBC and mouse alloantigens. TH2 cells were found among clones specific for mouse alloantigens, fowl gamma-globulin, and KLH. The relationship between these two types of T cells and previously described subsets of T helper cells is discussed.
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PMID:Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. 241 30

Borrelia burgdorferi strains (six isolates from North America and 28 isolates from Europe) were analyzed by physicochemical and immunological methods. By SDS-PAGE, all Borrelia burgdorferi strains tested had two major proteins with constant molecular weights of 60 and 41 kDa and one, two, or three variable low molecular weight proteins (OspA = 30-32 kDa, OspB = 34-36 kDa, pC = 21-22 kDa). All combinations--except OspB alone or OspB/pC--were observed. Borrelia burgdorferi strains were different from relapsing fever borreliae by strong reactivity with OspA- and/or pC-specific polyclonal antibodies, whereas relapsing fever borreliae were only weakly reactive. Among 25 Borrelia burgdorferi isolates, seven different serotypes of Borrelia burgdorferi were defined according to their reactivity in the Western blot with three monoclonal OspA-specific antibodies (H5332, H3TS, and LA5), four OspA- or OspB-specific polyclonal antibodies, and 12 polyclonal antibodies against whole borreliae. Antigenic differences between European CSF and skin isolates were observed, all skin isolates (n = 11) belonging to serotype 2 in contrast to only two out of seven CSF isolates. CSF isolates were antigenically heterogenous (serotypes 1, 2, 3, 4, and 5). Serotypes 6 and 7 were represented by two tick isolates, and the other European tick isolates are not yet fully characterized. Antigenic differences between European and North American strains may play a role in differences in the clinical picture of Lyme borreliosis.
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PMID:Antigenic variability of Borrelia burgdorferi. 246 Nov 35

Recombinant human GM-CSF has been expressed as a fusion protein in E. coli in the form of inclusion bodies. Using denaturing agents, acid cleavage and sulfitolysis, the biologically inactive GM-CSF protein could be highly purified and additionally renaturated under suitable reoxidizing conditions. The thorough repair of the two disulfide bridges could be confirmed by sequencing fragments obtained by tryptic digestion. Refolding of the molecule has been studied by CD spectrometry and identity by Western blotting and SDS-PAGE analysis. As could be demonstrated, full biological activity (colony-forming assay with fresh human bone marrow cells) was restored during renaturation of the GM-CSF protein. Further proof of biological equivalence of the E. coli-derived protein with a yeast-derived biologically active rh GM-CSF has been published elsewhere.
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PMID:E. coli derived human granulocyte-macrophage colony-stimulating factor (rh GM-CSF) available for clinical trials. 246 52

The purpose of this study was to evaluate the effect of various therapeutic regimens on: 1) intrathecal IgG synthesis on the basis of IgG Index value, 2) oligoclonal IgG spectrum visualized by SDS-PAGE of unconcentrated CSF, 3) CSF antibody specific activity against MBP estimated by solid phase RIA and expressed in cpm/micrograms IgG, and 4) immune complex (CIC) level in the CSF estimated by C1q binding solid phase RIA. CSF antibody against Gal-C and ganglioside was also estimated. Patients with clinically definite MS were selected according to 4 therapeutic regimens: group 1, subjected to Mega-dose prednisone therapy (4000 mg over 54 days), group 2, subjected to moderate dose prednisone therapy, group 3 subjected to Mega-dose Solu-Medrol therapy (7500 mg over 10 days), and group 4, subjected to intravenous Cyclophosphamide therapy (4000 mg over 10 days). This last group was characterized by chronic progressive course of disease. Intrathecal IgG production was significantly reduced in all 4 groups as a result of therapy. More pronounced reduction was obtained in Mega-dose prednisone (p below 0.001) and CY (p below 0.001) treated group. Therapeutic regimens did not influence the IgG oligoclonal pattern. The moderate dose prednisone therapy and Mega-dose Solu-Medrol therapy on CSF IgG anti-MBP antibody specific activity were less effective than the Mega-dose prednisone medication. CY therapy did not influence anti-MBP antibody specific activity in MS group characterized by chronic progressive course of disease. The influence of therapeutic regimens on elevated CIC level in the CSF was insignificant. In our study CSF the anti-galactocerebroside antibody appeared to be of IgM class.
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PMID:Effect of immunosuppressive therapy on humoral immune response in multiple sclerosis. 248 26

A growth factor acting synergistically with IL-3 on thiol-sensitive "mucosal type" bone marrow-derived mast cell lines, and therefore termed mast cell growth enhancing activity, is present in PWM stimulated spleen cell conditioned medium. Mast cell growth enhancing activity can be partially purified and completely separated from IL-3, IL-4, and IL-5, and for the most part from IL-6 and GM-CSF using strong cation exchange and Procion red affinity chromatography. Mast cell growth enhancing activity binds to Con A-Sepharose and can be digested with trypsin and chymotrypsin. It shows a Mr ranging from 37 to 43 kDa under nonreducing SDS-PAGE and a main isoelectric point ranging from 6.2 to 7.3.
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PMID:Partial purification of a mast cell growth-enhancing activity and its separation from IL-3 and IL-4. 278 57

The results from the Danish model of acoustic neuroma surgery are presented. In the period from 1976 to 1985, 300 patients with acoustic neuromas were operated upon using the translabyrinthine procedure. Only one small intrameatal tumour was encountered; 96 tumours were medium sized and 203 were larger than 25 mm. Of these 118 measured more than 40 mm. Mortality rate was 2%, CSF leaks occurred in 11%, and had to be closed surgically in 5%. Facial nerve function was postoperatively normal in 66%, slightly reduced in 17%, moderately reduced in 8% and abolished in 9%. Reconstruction, most often as a XII-VII anastomosis, was performed in only 6% of the patients. Cerebellar symptoms, which occurred in 45% preoperatively were present in only 7% after surgery. The preoperative hearing in both the tumour and non-tumour ear was analysed in 72 patients with tumours smaller than 2 cm. In the tumour ear, only four patients had a PTA of 0-20 dB and SDS of 81-100%; eight patients had a PTA of 0-40 dB and SDS of 61-100%; 14 had a PTA of 0-50 dB and SDS of 51-100%. This means that only a maximum of 5% of the patients, using the broadest criteria, could be candidates for hearing-conserving surgery. In all these patients the contralateral ear had hearing within normal limits (PTA 0-20 dB and SDS 95-100%). Since preservation of hearing would be achieved in only half of those subjected to suboccipital removal and since the hearing retained in patients with successful operations generally is poorer than the preoperative level, the number of patients obtaining serviceable hearing is so modest that preservation of hearing cannot be considered a valid argument in favour of suboccipital tumour removal. From a statistical point of view the risk of losing hearing in the opposite ear after tumour removal is negligible. The general morbidity after suboccipital surgery is higher than after translabyrinthine surgery, and hearing loss must be listed low among the other sequelae after tumour removal.
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PMID:Acoustic neuroma surgery: results of translabyrinthine tumour removal in 300 patients. Discussion of choice of approach in relation to overall results and possibility of hearing preservation. 262 90

Colony-stimulating factors (CSFs) produced by two simian virus 40(SV40) transformed macrophage cell lines (BAM1 and BAM3), and three hybrids (HM3-11, HM3-12, and HM3-14) derived from fusion between BAM3 and a Chinese hamster cell line (hs222-16) were examined. HM3-11 and HM3-14 produce two molecular species of CSF, which are not found in the conditioned media from cultures of BAM1 and BAM3 or lipopolysaccharide (LPS), phorbolmyristate-acetate (PMA), and zymosan-stimulated BAM3. HM3-12, which is classified into another group in terms of CSF secretion, does not produce these two CSFs. On the basis of various criteria, one of these CSF species (peak 1-CSF) was characterized as a macrophage-colony-stimulating factor (M-CSF). The other CSF (peak 2-CSF) induced a group of bone marrow cells in granulocytes and macrophages as well as growth of a mast cell line, IC2. This CSF has an apparent molecular weight of 18,000, estimated by SDS-polyacrylamide gel electrophoresis. Unlike interleukin 3 (IL3) from WEHI-3 cells, the growth factor activity of peak 2-CSF binds to DEAE-Sephacel. Thus, peak 2-CSF is similar to a granulocyte-macrophage colony-stimulating factor (GM-CSF) rather than to IL3. The anti L cell CSF serum does not inhibit the CSF activity in Chinese hamster fibroblast conditioned medium, and the IC2 cells do not respond to Chinese hamster lung conditioned medium (CHLCM), suggesting that peak 1- and peak 2-CSF are of mouse origin.
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PMID:Properties of colony-stimulating factors produced by macrophage cell lines and hybrid cells. 302 9

Analysis of human serum reactivities to antigenic components of soluble Taenia solium metacestode proteins showed the predominant presence of determinants shared by T. solium, Echinococcus multilocularis and E. granulosus. Two polypeptides were demonstrated by SDS-PAGE and Western blot or enzyme-linked immunoelectrotransfer blot (EITB) assay to bind serum and CSF antibodies only from T. solium cysticercosis patients. These species-specific antigenic polypeptides focused between pH 4.6 and 3.9 after resolution by isoelectric focusing followed by EITB. The high species-specificity demonstrated by the present techniques offers the opportunity to confirm serologically an infection by T. solium metacestode.
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PMID:Demonstration of species-specific and cross-reactive components of Taenia solium metacestode antigens. 308 29

The normal myeloid hematopoietic regulatory proteins include one class of proteins that induces viability and multiplication of normal myeloid precursor cells to form colonies (called MGI-1 = CSF or IL-3) and another class (called MGI-2 = DF) that induces differentiation of normal myeloid precursors without inducing cell multiplication. Different clones of myeloid leukemia cells can differ in their response to these regulatory proteins. The present experiments characterize proteins secreted by Krebs ascites carcinoma cells that induce differentiation of 2 different types of myeloid leukemic cell clones (clones II and 7-M12). The results indicate the following: (1) Krebs cells produce 2 distinct and separable proteins, each inducing differentiation in one of the leukemic clones. (2) One protein induced differentiation of clone-II myeloid leukemic cells and of normal myeloid precursor cells was free of any colony-inducing (MGI-1 = CSF or IL-3) activity, bound to double-stranded mammalian DNA, and was thus a differentiation-inducing protein MGI-2. This MGI-2 protein (MGI-2A) was purified to a single silver-stained band on an SDS polyacrylamide gel. (3) The other protein induced differentiation of clone 7-M12 myeloid leukemic cells, did not bind to double-stranded DNA and could not be separated from the myeloid growth-inducing protein MGI-1GM (GM-CSF) after 6 steps of purification including high-pressure liquid chromatography. The use of specific antisera confirmed that the protein which induced differentiation of clone 7-M12 leukemic cells was MGI-1 GM. The results show that Krebs ascites tumor cells produce 2 different myeloid hematopoietic regulatory proteins that differ in their target specificity for different clones of myeloid leukemic cells.
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PMID:Target-cell specificity of hematopoietic regulatory proteins for different clones of myeloid leukemic cells: two regulators secreted by Krebs carcinoma cells. 325 91


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