Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.4.21.4 (trypsin)
42,187 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Arterial endothelial cells were obtained from bovine aortae by mild treatment with collagenase and medium perfusion. These cells were cultured in RPMI-1640 medium containing 15mM Hepes buffer and 35% fetal calf serum at pH 7.35. Essentially all (90-95%) the effluent cells were viable and 80% of these cells attached to the substratum within 1 hour. Small patches of attached cells coalesced to form confluent monolayers in 3-5 days. Confluent monolayers of endothelial cells consisted of a homogeneous population of tightly packed, polygonal cells. Selected cultures were serially subcultured (trypsin-EDTA) for 12-14 months (30-35 passages) without any apparent change in morphology or loss of growth characteristics. Primary and three-month old (15 passages) cultures had population doubling times of 32-34 hours and 29-31 hours, respectively. These cells (primary and subcultures) did not require a minimum cell number to become established in culture. Bovine endothelial cells (primary, first, fifth and thirteenth passages) were characterized ultrastructurally by the presence of Weibel-Palade bodies, pinocytotic vesicles and microfilaments and immunologically by the presence of thrombosthenin-like contractile proteins and Factor VIII antigen. The intercellular junctions of post-confluenct cultures stained specifically with silver nitrate. From these data, we concluded that identifiable endothelial cells could be obtained from bovine aortae and cultured and maintained for prolonged periods of time.
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PMID:Culture of arterial endothelial cells: characterization and growth of bovine aortic cells. 17 37

The effect of human peripheral blood polymorphonuclear leucocyte (PMN) extracts and PMN granule lysates on in vitro immunoglobulin (Ig) synthesis by autologous peripheral blood mononuclear cells was studied. The mononuclear cells were cultured for 3 days with or without autologous plasma. Newly synthesized Ig in the culture supernatants was measured using 14C-labelled amino acids by an immune coprecipitation method. Upon addition of a PMN extract to plasma-free cultures Ig synthesis was stimulated, the mean stimulation index (SI) of cultures from thirteen individuals, including nine normals, three patients with rheumatoid arthritis and one with psoriatic arthritis being 1-8 +/- 0-2 in comparison with control cultures (P less than 0-05). By contrast, in 10% fresh autologous plasma, PMN extracts yielded a mean SI of 0-9 +/- 0-1 indicating inactivation of the active extracts by plasma inhibitors. In experiments using PMN granule lysates containing high concentrations of beta-glucuronidase and cultured in RPMI 1640, the mean stimulation index was 3-2 +/- 0-7. Stimulation of Ig synthesis was also produced by trypsin. Stimulation of Ig synthesis was also produced by trypsin. Stimulating factors in PMN extracts were inhibited by Trasylol, a protease inhibitor. These results indicate that trypsin and proteolytic lysosomal enzymes in PMN increase Ig synthesis of human peripheral blood mononuclear cells. They suggest a possible new role of PMN in the potentiation of immunoglobulin synthesis.
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PMID:Enhancement of in vitro immunoglobulin synthesis of human lymphocytes by lysosomal enzymes from polymorphonuclear leucocytes. 30 Mar 11

The binding of human IgE myeloma proteins to 16 human cultured lymphoblastoid cell lines was studied by measuring specific uptake of radiolabeled deaggregated IgE myeloma proteins and/or E-IgE rosette formation. Eight lines, RPMI-8866, Wil-2WT, RPMI-6410, RPMI-1788, RPMI-4265, Clowers, COLO-59 and Victor, bound IgE as shown by at least one of these methods. The lines, RPMI-4098, SCRF-5004, NC-37, Daudi, Raji, P3JHR-1, RPMI-1301 and Molt-4 did not bind IgE. Of the positive cell lines, 58 to 98% of the cells formed E-IgE rosetts. The binding of IgE was Fc fragment specific. It could only be inhibited by human IgE and its Fc fragment but not by IgE Fab fragments and Ig of other classes. The binding of IgE also appeared to be species specific, since a rat IgE myeloma protein did neither bind to the cells nor inhibit the binding of human IgE. The binding of IgE was relatively temperature independent and was abolished by trypsin and pronase pretreatment of the cells. Most of the cell lines binding IgE did not bind IgG but had surface immunoglobulin and did not form spontaneous E rosettes. These data suggest that certain lymphoblastoid cells may have receptors for IgE.
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PMID:Binding of IgE myeloma proteins to human cultured lymphoblastoid cells. 79 32

The secondary cyst tissues derived from mice infected with protoscoleces of Echinococcus granulosus for 8-10 months were digested with 0.25% trypsin at 37 degrees C for 30 min. The separation of different cells in the remaining suspension was achieved by discontinuous gradient centrifugation. The germinal cells were washed 3 times with ice-cold HBSS, and then cultivated in the medium of RPMI 1640 supplemented with 20% of calf serum. The cells were kept in an incubator at 37 degrees C in an atmosphere of 95% air-5% CO2. After incubation for 5-7 days, the germinal cells began to multiply accompanied by the enlargement of cells as compared with those before incubation. The surface of both isolated and/or cultured cells showed smooth appearance examined by scanning electron microscopy. Immunofluorescence assay and enzyme-linked immunosorbent assay had been used for examining the specific antigenicity of the cells. The results showed that antigen components of E. granulosus were detected either on cell surface or in soluble proteins of the cells. Furthermore, 120 NIH female mice were inoculated intraperitoneally with 1-5 x 10(7) cultured germinal cells and sacrificed 1-3 months after inoculation. Only 2 cystic materials had been detected in two mice. Of which, one located in the liver and the other in peritoneal cavity of the animals. Histological examination noted that the cystic materials consisted of germinal layer and cyst fluid, but no laminated layer was observed. The above mentioned evidence demonstrated that the cells isolated from the cysts of E. granulosus were germinal cells.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:[Isolation of germinal cells from the secondary cysts of Echinococcus granulosus harbored in mice]. 139 14

Cell motility has been associated with metastatic ability in the Dunning R3327 rat prostatic adenocarcinoma model. Cancer cell motility promoters but not inhibitors have been described by many investigators. Serum-containing and serum-free media conditioned by the nonmotile, nonmetastatic G Dunning subline inhibited the motility of the highly motile, highly metastatic MAT-LyLu subline. Motility inhibition by the G subline-conditioned serum-free media was lost upon heating to 100 degrees C and by treatment with trypsin. The motility inhibitory protein(s) had a molecular weight exceeding 50,000 as determined by diafiltration. G subline-conditioned RPMI 1640 contained several proteins with molecular weights of between approximately 53,000 and 116,000 when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. One of these bands may represent the first inhibitor of cancer cell motility identified.
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PMID:Cancer cell motility-inhibitory protein in the Dunning adenocarcinoma model. 155 38

A primary culture method was established by comparing the different effects of four methods of enzymatic separation--trypsin, collagenase with and without trypsin pretreatment, and a trypsin-collagenase mixture--and five media: DMEM, DMEM and Ham's F 12 mixture, F 12, RPMI 1640 and Medium 199. The trypsin pretreatment/collagenase method was most preferable considering the high number of isolated cells, satisfactory adhesion, good growth and a single population at subconfluence. DMEM and the DMEM/F-12 mixture resulted in the best adhesion, cell growth and cell number at confluence. Primary cells separated by the trypsin pretreatment/collagenase method and cultured in DMEM were responsive to parathyroid hormone at the proliferating stage and had higher alkaline phosphatase activity than cells cultured from gingiva and mucosa after reaching confluence. The long-term cultured cells formed nodules that were slightly mineralized. These results indicate that the cultured pulp cells had properties characteristic of pulp cells in vivo. This enzymatic separation method may be useful in studies of the regulation of pulp metabolism and odontoblast differentiation.
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PMID:Establishment of primary cultures of pulp cells from bovine permanent incisors. 166 Feb 58

When isolated from lysates of the RPMI 8866 B lymphoblastoid cell line, the CD23 antigen was found to be present be in two forms corresponding to the intact 45-kDa membrane species and to a 25-kDa product that is more usually found as a released fragment in the extracellular medium. By contrast with preparations of extracellular species of CD23 which were seen to be variable in their biological activity, cell-associated CD23 was consistently mitogenic both for autogenous transformed B lymphoblasts and for pre-activated normal B cells. Addition to the normal cocktail of protease inhibitors of N alpha-tosyl-L-lysine chloromethylketone (TLCK), which has selectivity for trypsin-like serine proteases, resulted in preparations of CD23 from RPMI 8866 cell lysates that were exclusively in the 45-kDa intact form; such material retained full and reliable activity in the biological assays. The implications of these observations for the autocrine control of B lymphocyte growth are discussed.
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PMID:Intact, 45-kDa (membrane) form of CD23 is consistently mitogenic for normal and transformed B lymphoblasts. 213 59

Primary rhesus monkey kidney (MK) cells have long been the cells of choice for isolation and propagation of the human paramyxoviruses (parainfluenza 1, 2, 3, 4A, 4B, and mumps). However, problems with the supply and cost of MK cells and the presence of endogenous viruses, including herpes B virus and SV-5, necessitated a search for an alternative cell line. Continuous cell cultures of human origin (L132, A-549, HuT-292, HEK, G-293, G-401, A-498, A-704, CAKI-1, RD) and simian origin (LLC-MK2, BSC-1, MA-104, Vero) were evaluated for their capacity to support the growth of the human paramyxoviruses, as followed by cytopathic effect, hemadsorption, hemagglutination, and EIA. NCI-H292 (HuT-292) human lung mucoepidermoid carcinoma cells (ATCC # CRL-1848) proved to be the most sensitive line for cultivating all serotypes and strains of the paramyxoviruses. These cells were also shown to be a suitable substitute for MK in primary isolation of paramyxoviruses from clinical specimens. RPMI-1640 with 1.5 micrograms/ml trypsin was the preferred maintenance medium; alternatively, Eagle's MEM supplemented with 1.5 micrograms/ml trypsin and 0.1% ITS was satisfactory. NCI-H292 cells are a continuous line with excellent growth characteristics, although the genetic polyploidy of the cells may limit the number of passages of usable cells.
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PMID:NCI-H292 as an alternative cell line for the isolation and propagation of the human paramyxoviruses. 226 Sep 24

This study indicates that human IgE-binding factors (IgE-BF) found in the cellfree culture supernatant (CSN) of Fc epsilon R-bearing B cells are breakdown products of the surface Fc epsilon R. This conclusion is suggested by the following observations. 1) Fc epsilon R and IgE-BF share several antigenic determinants as shown by immunoprecipitation with several Mab to Fc epsilon R (MabER) and SDS-PAGE analysis of the precipitates. 2) Upon incubation at 37 degrees C, normal tonsillar lymphocytes lose their Fc epsilon R and this is associated in a time-related manner with the release in the CSN of molecules reacting with two MabER. 3) Surface radioiodinated tonsillar lymphocytes or RPMI 8866 cells release labeled IgE-binding molecules displaying the same antigenic composition and the same migration on SDS-PAGE as purified IgE-BF. 4) Peptide mapping of highly purified IgE-BF and Fc epsilon R reveals the presence of several identical fragments after digestion with either alpha-chymotrypsin, trypsin, or papain. Moreover, papain digestion of the 25-27 kD IgE-BF and of the affinity-purified Fc epsilon R, generated a 15 kD fragment reacting with two MabER and that is known to bind IgE. Although these data strongly suggest that IgE-BF may be directly derived from cell surface IgE receptors, they do not exclude the possibility that some IgE-BF may also be secreted without being first anchored in the cell membrane.
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PMID:Relationship between human IgE-binding factors (IgE-BF) and lymphocyte receptors for IgE. 243 96

It has been reported that the 45-kDa low affinity Fc epsilon R (Fc epsilon R2) on B cells is cleaved spontaneously from the cell surface to release a 28-kDa soluble fragment (sFc epsilon R2). This study demonstrates an additional mechanism by which B cells generate this fragment. Data from 35S methionine pulse-chase experiments with the Fc epsilon R2 bearing human B lymphoblastoid cell line, RPMI 8866, and immunoprecipitations of cell lysates and culture supernatants with an Fc epsilon R2 specific mAb, mAb 25, demonstrates the existence of a cell-associated 28-kDa Fc epsilon R2 fragment. This fragment was shown by partial amino(NH2)-terminal sequence analysis to be identical to the previously described 28-kDa sFc epsilon R2. The resistance to cell treatment with trypsin indicated that it was located intracellularly. Its appearance early in the biosynthesis of the Fc epsilon R2 (within a 10-min pulse), before the Fc epsilon R2 reached the cell surface, suggested that some of this fragment was generated intracellularly. Neutralization of acidic organelles with NH4Cl inhibited the formation of this intracellular fragment, strongly suggesting that it was a produce of intracellular cleavage of the Fc epsilon R2. Finally, this 28-kDa intracellular fragment was shown to be released into the culture supernatant, suggesting an intracellular mechanism by which the cells generate sFc epsilon R2.
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PMID:Intracellular cleavage of newly synthesized low affinity Fc epsilon receptor (Fc epsilon R2) provides a second pathway for the generation of the 28-kDa soluble Fc epsilon R2 fragment. 252 83


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